(Biological Optical Technologies)

Dedicated to Mainstreaming Live-Cell Microscopy into
Medical Research and Clinical Practice

The products contained in this catalog were developed to meet the demanding
requirements of the current renaissance in microscopy. This revolution in microscopy
includes: confocal imaging, standing-wave microscopy, multi-probe fluorescencemicroscopy, computerized imaging, movie loops, etc.
Live-cell microscopy environmental control systems require careful planning and attention
to every detail. This catalog has been prepared to assist in the selection of a variety ofadvanced products and technologies designed specifically for the live-cell microscopist.
In addition to being a resource of technologies for the experienced microscopist, it canbe used as an informative guide for those just entering the exciting and demanding worldof live-cell microscopy. We wish you much success in your work and hope you find our
products and support beneficial. If you do not find the live-cell micro-observation products
you need in this catalog, please call us and discuss your specific problem. We would be
happy to share our knowledge base with you.
Bioptechs can be reached Monday through Friday, 9:00 AM to 5:00 PM Eastern Time.
All calls are answered by a competent human being. We will never use those annoying
automated phone systems during business hours!!!

Voice

Toll Free 877-LIVE-CEL (548-3235) U.S. and Canada
724-282-7145 (Sales and Technical assistance)Fax

724-282-0745 (24 hours a day)

E-Mailinfo@bioptechs.com (Checked regularly)Web

http://www.bioptechs.com (Includes automatic E-mail)1

What is Bioptechs?
Bioptechs is an innovative American optical engineering company that designs, develops, and manufactures
live-cell microscopy environmental control instrumentation for qualitative and/or quantitative light microscopy.
Our products are dedicated to the science of live-cell microscopy and include imaging systems, microobservation environmental control instruments, perfusion pumps, bio-compatible tubing for cell culture, andspecialized fittings and adapters as well as numerous supplemental items. Bioptechs has developed andpatented several technologies which are indispensable to the live-cell microscopist which include:• Microaqueduct laminar perfusion in a temperature-controlled optical cavity• A unique symmetric closure mechanism for optical cavities• Objective thermal regulation devices to reduce temperature gradients for high N.A. objectives• A unique hybrid culture dish system specifically designed for live-cell microscopy which utilizes anadvanced technique of first-surface thermal transfer to quickly, accurately, and uniformly maintainthe thermal, optical, and fluid requirements of cultured cells and tissue
Bioptechs products and techniques have demonstrated their ability to out-perform traditional methodologies
and have become the established standard method of micro-environmental control in many leading
universities and pharmaceutical companies. Bioptechs manufactures more than 80 standardized products
and provides customers with the availability of a variety of related products and services.
Mission:
Man is a highly complex biological structure whose composition is being studied at every level from molecular
structure to behavioral science. Within this range lies the area of cell biology. Until the past ten years, mostof the microscopy of cells has been done with fixed cells. The information gathered has been limited to onlythat which can be derived from the dead remains of cells. Now scientists have a new set of tools available
to them, such as highly sensitive electronic cameras interfaced to computers and microscopes that areable to optically detect minute traces of specialized dyes or indicators. This instrumentation can provide
both visual and quantitative information about processes and behaviors within the cell that were previously
unobtainable. Bioptechs recognizes the importance of the ability to reproduce the cells’ host environmenton the microscope, without sacrificing optical functionality, as a critical component of modern biologicalexperimentation. Our systems are used primarily in basic research applications. We are also developing
new protocols for the analyzation of live primary cells obtained from human patients to determine the mostappropriate treatment as a routine method of health care. Bioptechs’ mission is to develop and manufacture
micro-environmental control systems to enable advancements in live-cell imaging for research and clinicalapplications.
Bioptechs products:
All Bioptechs’ products are designed, manufactured, and tested at the Bioptechs’ facility in Butler, PA, and arebacked by a one-year warranty. Bioptechs live-cell systems provide full optical compatibility with all modes oflight microscopy. Bioptechs’ controllers feature a real-time temperature display and an ultra-fast learning curve
to compensate for temperature changes due to surface evaporation or perfusion. The standard controllers
have a special out-of-limit alarm circuitry to ensure the safety of the specimens and a temperature range
from ambient to 50°C. Extended ranges from ambient to 100°C are available upon request. Our temperature
control systems are economically adaptable to a variety of critical temperature control applications. Bioptechs
also offers opto-electronic and opto-mechanical design services specializing in light microscopy. Our inhouse machine shop affords the luxury of making modifications or prototyping new designs quickly andeconomically.
Applications include but not limited to:
AIDS researchAnesthesiology
Apoptosis
Artificial membrane
Avian research
Bacteriology
Brain slice

Calcium ratioing
Cancer cell migration through tissue
Cardiac myocyte recording
Chemo assay of cancer patient tumor cells
Developmental biology
Drug testing & development
Induced change assays
2

In vitro fertilization (IVF)
Morphology
Muscle force transduction
Neuronal stimulation & recording
Soft tissue microscopy
Tissue engineering
Toxicology

Contents
Observation of Live Cells on the Light Microscope

4-9Open Dish Systems (Delta T Culture Dish System)

14 - 26

Options for the Delta T Culture Dish System

16 - 26

Membrane Micro-Observation

20

Brain Slice Micro-Observation

21

Open System Perfusion

22

Delta T Stage Adapters

23

Accessories

26

Closed Chamber Systems (FCS2 System)

27 - 29

Closed System Perfusion (FCS2)

30

Options for the FCS2

32

Cooled FCS2

33

FCS2 Stage Adapters

34

FCS3 Upright Closed System Chamber

35 - 37

Closed System Perfusion (FCS3)

38

Objective Warming

39

Objective Cooling40

Perfusion Systems

41

Dual Pump Control

43

Perfusion Tubing

44

Reference Information

45

System Profile Guide

46

Checklist for Closed System Chambers

47

Checklist for Open System Chambers

48

How to Size Your Objective

49

Price Lists

50 - 52

User References

53 - 543

Observation of Live Cells on the Light Microscope
Reproduced and edited from an invited article appearing inCells A Laboratory Manual,Volume 2, pg 75.1 - 75.13.
Daniel C. Focht, David L. Spector, Robert D. Goldman, Leslie A. Leinwand

Introduction

microscopes did not have the image-contrasting enhancementsthat modern microscopes have. Therefore, the live-cellmicroscopy that was done during the film era was either poor bytoday’s standard or limited to high-contrast specimens.

The direct observation of the dynamic properties of known
molecular species in vivo has been made possible through avariety of molecular, biochemical, immunological, and imaging
techniques. For example, the now widespread use of GFP tags
permits the direct observation of specific cellular constituents invivo. However, in order to observe live and healthy mammaliancells in the microscope field, there are technical problems
with respect to the maintenance of temperature, pH, etc., of
the culture medium. This is not a significant problem whenobserving non-mammalian or plant cells. Typically, live-cell
experiments can be classified into two categories; developmental
studies to establish natural behavior, or induced change to study
the effects of a controlled factor. The correlation between invitro and in vivo phenomena is paramount for mammalian livecell investigations. Therefore, it is of the utmost importance tosimulate the host conditions of the isolated specimen accurately
during live-cell microscopy. This chapter describes multiple
approaches for the micro-observation of live cells.

Recent Technical Developments

The following developments have made quantitative live-cellmicroscopy easier:• The development of image-contrast enhancement techniquessuch as, DIC, Hoffman, Phase, Varel, Fluorescence, Multi
Photon, and Confocal Microscopy• Video cameras and quantitative CCD cameras capable of
low-light detection and electronic contrast stretching• Desktop computers bringing quantitative imaging capabilities
into mainstream science• Advances in the development of dyes and labeling techniques• Micro-environmental control systems developed to keep pacewith the associated technologies
Today these technologies have provided the instrumental
foundation for conducting live-cell experiments, requiring lesstime for engineering equipment, and leaving the researcher moretime for biology (Pentz and Schulle 1981; Webb 1986; Shotton1988). Cells can be maintained in an incubator, but putting the
microscope in an incubator is impractical. The logical extension isto duplicate incubator conditions on the stage of the microscope.
Cells need certain known regulated conditions such as nutrients,
removal of waste products, carbon dioxide, a growth surface
matrix or lattice in the case of tissue, and temperature control. In
order for cells to be visualized, they must be in an environmentthat is both conducive to their viability and compatible with themicroscope’s optical limitations.

History

Specimens used for early microscopy were often alive, or atleast temporarily alive. Techniques for fixing and staining
were developed along with advances in microscopy. As
microscope optics improved, the limitations of the opticalsystem defined the necessity for specimen preparation.
Microscopists played a game of give and take based on opticalconstraints. Magnification, numerical aperture, resolution limits,
working distance, and correction for chromatic aberration
were recognized as interrelated factors. In addition, methodsof illumination to include bright-field, dark-field, phase, andfluorescence were brought into mainstream microscopy. With
these forms of microscopy available, it was accepted that the
specimen needed to be prepared to meet the optical limitationsof the microscope. These preparations most often required
fixing the specimen. Cameras using film were combined withmicroscopes to document images. Conventional film-based
photomicroscopy was suitable for most bright-field, dark-field,
and fixed fluorescence images because exposure time was of noconcern. Time lapse images of live cells were typically acquired
using bright-field modes because the low sensitivity of film
required high light levels. Live-cell fluorescence movies were
not possible due to the toxicity of the dyes and phototoxic effectof long exposures. Although fixed cells yield less informationin comparison to live specimens, there are some advantages.
Fixed cells are flatter and last longer. They don’t move or
require constant care, and they can be stained with a varietyof compounds to selectively enhance contrast. Most anti-fadereagents are compatible with fixed (but not live) cells. When
images of live cells were recorded on conventional film, themicroscopist was limited by the sensitivity and resolution of
the film, which also limited the kinds of experiments that could
be done. Film has many limitations when compared to modern,intensified, or cooled CCD cameras, and photomultiplierscanning techniques. Consequently, early live-cell chambers
were inherently limited by the associated technology. Early

Micro-Environmental Chambers

There are two basic forms of specimen containment and microenvironmental control. In applications where single-cell analysis
is appropriate, the specimen must be contained in an opticalenclosure. The two most commonly used methods of microobservation are the open culture dish in which fluids are open to
the atmosphere, and the closed system, which is sealed.
Typical open culture dish systems are the simplest form of
live-cell observation and are generally well suited for shortterm experiments or applications where physical access to thecells is necessary. The traditional method of growing cells ina container, then observing the cells on a peripherally heatedmicroscope stage, is adequate for investigations requiring only
basic modes of relatively low-resolution microscopy. For moreadvanced investigations requiring high-resolution imaging using
fluorescence, DIC, 2 Photon, and/or confocal modes, a moreadvanced environmental control system should be used. Various
types of simple culture chambers were developed for observing
live cells under the microscope. Some of the more popular types
include the Maximow Chamber, the Romicron Chamber, andthe Rose Chamber. The Maximow Chamber was the simplest
early chamber, consisting of a thick micro-culture depression
slide made of heavy glass. An earlier version of a closed system

4was constructed by placing a gasket between a coverslip and aslide or two coverslips. The Romicron chamber is also made ofplate glass, but with an optically ground cylindrical depression
centered in the slide so that the bottom of the depression is
optically clear. A double-coverslip technique is used to cover thechamber. This consists of a larger, usually rectangular, slide uponwhich is placed a smaller circular coverslip with the primary
explant or cells of interest in medium. The smaller coverslip isheld onto the larger slide by capillary action. This double coveris then inverted to span across the cylindrical chamber and sealed(typically with beeswax). This sort of chamber is appropriate
for use with experiments where perfusion is not needed and theexperiment is of short duration so that pH, CO2, temperature,and the buildup of waste products are not common problems.
The Rose Chamber is a more elaborate version of a simple
closed chamber, consisting of a thick plate glass slide having acircular opening through the slide to form the chamber. The top
and bottom of the chamber are sealed, typically with coverslips
placed above and below the opening through the slide, often witha rubber O-ring or gasket between the slide and a coverslip toallow perfusion into and out of the chamber. This sandwich ofcoverslip, O-ring, slide and coverslip is held together by metalrims on the top and bottom of the sandwich that may be screwed
together to form a tight seal. Although these simple chambers
may be suitable for many purposes, the more sophisticatedchambers described in this chapter generally provide a moreadvanced environmental control system appropriate for more
complex experimental observations.

warm air blowers and/or infrared lamps surrounding the stage.
This is because these devices involve maintaining temperatureby switching heaters on and off continuously, frequently causing
focal drift and contamination. An alternative solution to the above
techniques utilizes first-surface thermal transfer to regulate celltemperature. An optical cavity or coverslip can be temperature
controlled by passing a current through an electrically conductive
coating on one of the optical surfaces that defines the opticalcavity. This technique has been proven to have many advantages
over traditional methods.
The following is a list of some of the typical difficulties
encountered with traditional chambers:
•
•
•
•
•
•
•
•

Coverslip breakage that occurs during assembly
Leakage onto and sometimes into the microscope
Lack of flexibility to accommodate varied volumetricrequirements
Fixed flow characteristics
Flow rates slower than biological events to be recorded
Minimal volumes larger than required by protocol
Inadequate temperature control
Lack of compatibility with all modes of microscopy

Selecting a Micro-Environmental System
Microscope Factors

Generally, inverted microscopes are used for cultured cells,
but in some cases, an upright microscope is appropriate. Whenusing an upright microscope, some concessions have to be madedepending on the intended modes of microscopy. For example,
special care must be taken in determining the magnification,
working distance, and numerical aperture, all of which are
related to the depth of field. It is also important to recognize thatfluid coupled objectives will need to be temperature controlled
when used with mammalian specimens. If a transmitted lightcondenser will be used, allowance must be made for the working
distance, physical size, and proximity to the chamber. The
geometry of the stage also must be matched or mated to theenvironmental chamber. Inverted microscopes are far morecompatible with live-cell experiments for the following reasons:cells gravitate downward, and the condenser can be employed
without interference from the stage. Upright microscopes have
two main disadvantages in open dish applications; the first being
the inability to fully utilize high N.A. objectives, and the otherbeing the position of the condenser to the specimen is limited by
the stage.

Why a Chamber?

A chamber can be any structure with optical, fluid, andtemperature control capabilities. This may be a simple culture
dish or flask, or a complex dedicated live-cell environmental
system.
• Observation of single cells in vivo with traditional diffraction
limited optics is difficult and sometimes impossible.
Therefore, cells have to be isolated from their host andstudied in an external optical culturing environment.
• The specimen needs to be studied on a flat transparent surface
to be compatibile with imaging.
• Cells also need a fluid containment structure for liquid media
of a suitable volume to maintain their viability.
• Perfusion may be needed to maintain control of pH, forremoval of waste products and to add reagents that will alter
cellular physiology.

Types of Environmental Control Techniques

Aside from the optical and physical embodiment of a chamber,a number of methods of temperature control are used. This isespecially important in studies of mammalian cells. Thermal
regulation devices employ a variety of techniques including:warmed airstream specimen warmers, electro-resistive heating
elements, Peltier effect, thermally controlled fluids, infrared
illumination, and first-surface thermal transfer. Temperature
control factors include: range, accuracy, speed, and recovery
time. Over the years, temperature has been controlled using
extreme measures, such as placing the entire microscope in
an incubator with only the eyepieces exposed. This is very
expensive and detrimental to the microscope. Another option
has been to encase the stage of the microscope with an acrylic
enclosure while pumping in heated air. The most popular option
is to use a heated plate or stage warmer to surround the culture
dish or coverslip chamber. The least desirable techniques are

Micro-Environment Physical Characteristics

When using an open system, the following characteristics need tobe addressed; volume, clear aperture, material, bottom thickness,geometry, evaporation and condensation, ambient light, view
angle, and biocompatibility. When selecting a closed system,
the following characteristics need to be addressed; fixed orvariable volume, separation of optical surfaces, perfusion-flow
characteristics, volume exchange rate, laminarity, shear stress,
and flow channel geometry.

5

Typical Culture Dish Observation

In applications where cells are being observed with low
numerical aperture (N.A.) objectives, an open culture dish andphase contrast or dark-field microscopy are frequently used.
Today’s sophisticated experiments, however, are far moredemanding on the cells’ environment. For example, during longterm developmental or induced-change studies, it is necessary
to accurately simulate the host conditions on the microscopestage without limiting the resolution of the microscope while
simultaneously maintaining the ability to change variables. This
simulation should include control of temperature, medium, pH,
and environmental toxicity. Medium and pH can be controlled
by regulated perfusion. The toxicity of the artificial environment
must be evaluated against compatibility with the specimen(Inoue 1986). Presuming the artificial environment is nontoxic or
biologically inert, one of the most difficult factors to control has
been temperature. With all the relevant papers on the importanceof accurate temperature control and its effect on data, it should
be readily apparent that temperature is a critical factor in most
mammalian live-cell experiments (Taylor and Wang, 1989;
Taylor et al. 1994). When using applications requiring high N.A.
objectives, most investigators use carefully supported coverslips
to attain optical compatibility and peripheral stage warmers tosimulate the host environment. This frequently means sacrificing
the accuracy of thermal control and/or adequate perfusion of thecells resulting in either inaccurate data due to compromised cells
or poor images. The characteristics and limitations of traditional
methods of open dish micro-observation are summarized here.
(See Figure 1)

Figure 1: Traditional open culture dish microscopy. The
traditional technique is limited by slow and inefficient stage
heaters, resulting in non uniform temperature distribution. In
addition, plastic dishes are poor conductors of heat. Thus, thetemperature does not recover quickly during or after perfusion.
Finally, the usable aperture of the dish is limited by the opening
in the heat-transfer plate.

Advances in Culture Dish Live-Cell Systems

An alternative solution to these problems is to use a microenvironmental system that utilizes a first-surface thermaltransfer technique. This technique utilizes a thin-film coatingof a nearly transparent, (see Figure 2) electrically conductive
coating applied to the bottom surface of a glass substrate,
which is then incorporated into a hybrid disposable culture dishstructure optimized for live-cell imaging. This technique provides
temperature control directly to the cells through a thermistor
feedback loop, which applies an electrical current through thecoated underside of the glass substrate (See Figure 3). The
thermal response using this technique is as fast as 0.1°C/sec.
Because of this fast response time, the controller can regulate thecurrent control within seconds, making it possible to compensate
for temperature changes that occur in the dish due to surface
evaporation, entropy, or perfusion. This fast response timeenables the addition of high-speed safety circuitry to protect thecells in the event of error. This technique offers high resolution
imaging capabilities through a uniform glass surface free ofstrain and is available in a variety of thicknesses, including
the popular No. 1.5 coverglass for high N.A. applications. The
dish environment is compatible with all modes of microscopy
including, but not limited to, bright-field, dark-field, phase, DIC,
fluorescence, reflection interference, 2 Photon, and confocal. The
first-surface thermal transfer technique on a coverslip, introduced
in 1993, provides the basis for many adaptations of the basic
principle. Since that time, numerous applications and accessories
have been developed. After becoming familiar with the accuracy,ease of use, and superior optical imaging obtainable with afirst-surface thermal transfer system, it is easy to envision andimplement numerous derivations for specific applications otherthan isolated cell culture. The following are examples of some ofthese uses, accessories, and benefits based on the Bioptechs Delta
T Culture Dish System.

Conventional culture dishes are made of plastic, which is known
to have the following characteristics:• Non uniform optical surfaces that degrade the image• Strain in the optical surface due to the injection molding
process that prohibits the use of optical systems employing
polarized light, including DIC• Thick bottom surface prohibits the use of high N.A. objectives• Inefficient heat transfer from the source to the cells which
results in long-term temperature-stabilization cycles.
Traditional culture dish warmers are peripheral heating devices
that must radiate heat through two inefficient thermallyconductive mediums (air and plastic), therefore resulting in:• Non uniform temperature distribution• Excessive thermal transfer time during thermal recovery (onthe order of minutes)• Thermal expansion of the metal surface causing verticaldisplacement which is apparent during 3D imaging or
confocal applications.
If these factors do not present a problem, a stage-warmerbased system is adequate and preferable. However, if theexperiment requires more accurate temperature control or opticalcompatibility, the techniques described in the following section
should be considered.

6Traditional Closed-System Chambers• Tissue slice specimens, natural or artificial, can be easilysuspended in an optical reference plane in the dish at theappropriate focal distance from the objective.
• Transmembrane migration or diffusion of cells or ions can be
dynamically observed during isolated perfusion of inhibitors
or changes of ion concentration to the basilateral surface.
• Continual perfusion can be employed to maintain low-volume
perfusion during long-term experiments.
• A coverglass lid is available for use during microscopy todefine an optical surface above the specimen and on thesurface of the liquid in the dish. When a series of images are
acquired in transmitted light mode, there is a changing fluid
interface between air and the surface of the medium, causing
image contrast shifts. Forming an optically flat surface on topof the medium eliminates this problem.
• An electrically heated, optically transparent lid eliminates
condensation from forming on the under surface of the cover.
• Cooling of the specimen can be accomplished by submerging
within the dish, a ring through which refrigerant fluid is
circulated.

A closed-system chamber is required when there is a need either
for complete isolation of the specimen from the outside world
or for the use of more advanced microscopic methods that areincompatible with the use of open dishes. In this case, cells areplaced in a temperature-controlled, perfusable optical cavity orplated onto a coverslip that is part of this cavity for microscopy.
There are several traditional configurations of these closedsystem chambers commercially available. Nearly all of themutilize the same basic characteristics. (See Figure 4 below)
Traditional closed-system chambers provide two optical surfaces
separated by a perfusion ring sealed with gaskets. This sandwich
is then clamped together by several other structures. With
this type of enclosure, the perfusion rate, volume, and opticalsuitability for various modes of microscopy are interrelated.
In most cases, perfusion with this configuration may result in
turbulence that can dislodge cells. The limitations of this design
impose a trade-off between functionality and compatibility withvarious modes of microscopy. Furthermore, temperature control
is usually achieved through the use of peripheral heaters or
warmed air blown across the stage. In either case, temperature
control is not reliable or maintained within an acceptable range
for critical experiments.

100.0

80.0

60.0

40.0

20.0

0.0

400

500

600

700

800

900

Transmittance (%) vs Wavelength (nm)When selecting a closed-system chamber, the following factors
need to be considered:• Optical compatibility for the intended modes of microscopy• Temperature control, including uniformity• Volume of chamber• Perfusion• Volume exchange rate for perfusion• Cell surface shear• Imaging aperture• Thermal effect of fluid coupled high N.A. objectives

Figure 2: Transmission spectra of Indium Tin Oxide coated0.170mm thick coverglass.

Figure 4: Traditional closed-system chamber.

Inherent Shortcomings of Traditional
Closed-System Chambers

Figure 3: First-surface thermal transfer.

• Excessive separation between optical surfaces limits
condenser N.A. in transmitted modes of microscopy• Fixed optical cavity volume• Requires lengthy fluid exchange rate
• Turbulent perfusion produces high cell surface shear-flow rate
• Not compatible with certain modes of light microscopy

Some advantages of the system are the highly accurate
temperature control, fast thermal recovery, and uniform
temperature distribution.

7

• Difficult to load and secure with cells• Prone to leakage

Advances in Closed-System Chambers

A solution to these problems is provided by the use of a
microaqueduct perfusion technique. A microaqueduct flow livecell, micro-observation chamber is prepared by incorporating
perfusion grooves into one of the optical surfaces that define the
Figure 5: Microaqueduct slide enclosure optical cavity, thereby
eliminating the perfusion ring common to most other chambers
and defining the optical cavity with only one gasket separating
the perfusion slide from the coverslip.
The physical configuration of these grooves produces a nearlaminar flow region in the optical cavity. The single gasket design
allows the user to define the size, volume, thickness, and shape ofthe optical cavity. In addition, microaqueduct perfusion

Bioptechs FCS2

provides large aperture flow inputs and outputs, eliminating
the problem of volume exchange rates. To further enhance theperformance of this design, the first-surface thermal transfer
technique (the electrically conductive, thermal controlled coatingon the microaqueduct slide) was incorporated, thus adding
thermal uniformity to the chamber even during experiments withperiods of no flow. This system is exemplified by the Bioptechs
FCS2 Closed System Chamber. (See Figure 5) Therefore, thespecimen, adherent cells on a coverslip, is maintained safely ina temperature-controlled optical environment that is compatible
with all modes of microscopy, including, but not limited to,
low and high N.A., transmitted, bright-field, dark-field, phase,
DIC, and reflected modes of fluorescence, as well as confocal.
For optimal temperature control when using high N.A. lenses(immersion), an objective lens heater is required. This system
is further characterized by a sophisticated temperature control
system easily modified for custom applications.

High N.A. Objectives

In high N.A. imaging, it should be noted that the opticalmedium used to couple the objective to the coverglass will act
as a heat sink. If the objective temperature is not regulated,
this will result in a temperature gradient across the field of
as much as 5°C. The microscope manufacturers have not yet
met the live-cell researchers’ needs by providing integrated
temperature control for the objective. So, an external objective
heater must be used. To control the temperature of the objective
accurately, it is necessary to overcome the constant drain of heatfrom any thermally conductive mass such as the nose piece or
microscope frame. The thermal characteristics of all objectives
and microscopes vary considerably, making it necessary to havean efficient transfer of heat to the objective and an intelligent
controller. Such a system would sense the temperature of theobjective at a point close to the specimen and regulate the heatapplied to the objective while taking into account the thermal
mass of the objective and the ambient conditions. A device forthis purpose, which particularly surrounds the upper portion of
the objective central tube with a heating band and measures thethermal transfer in a gap formed between the ends of the heating
bands, is exclusively available from Bioptechs, Inc. (Patent #5,410,429). The controller regulates the heater current in sucha manner that it maintains the temperature to within 0.1°C.
Although the objective heater is designed to fit most objectives,it does not fit all objectives. Some objectives, especially watercoupled objectives, are particularly difficult to work with. The
main problem is their large diameter and lack of adequate surface
area on which to transfer heat. Sometimes their size makes it
difficult, if not impossible, to have sufficient working room tofocus on the specimen. The microscope manufacturers haveagreed that heating of the objectives to 37°C will not harm theobjectives, but they have not made any comments about daily
heating. It is recommended that the objective be stored in a 37°C
enclosure when not in use to reduce the possibility of detrimental
effects of thermal cycling between physiological and room
temperatures.

Temperature Sensors
Heater Contacts
Perfusion Connections

Upper Gasket

Microaqueduct Slide (opticalsurface, perfusion guide andtemperature control)

Lower Gasket
Defines Flow Geometry

40 mm, #1.5
Coverglass
Self-Locking
Base with
Internal
Temperature
Control

Objective Heaterfor High N.A. Lenses

Perfusion

Perfusion is necessary for one or both of the following reasons;either experiments on live cells take place over a time span
greater than cells will tolerate without fresh media, or theexperiment is one in which chemical change is induced throughperfusion. In both cases, it is necessary to have a method of
perfusing cells in their micro-observation environment thatwill not impede the experiment or the acquisition of electrical,

Figure 58chemical, or optical data. Due to the small fluid volume in livecell chambers, the diaphragm effect produced at the unsupported
aperture of the coverslip, and the sensitivity to shear stressesof the cells, selection of a perfusion system requires great careand, in some cases, a trade-off. The ideal perfusion pump would
provide a closely controlled, purely analog flow indefinitely. Thisis currently not available. The three basic methods of perfusion
are gravity flow, manual injection with a syringe, and mechanicalpumps. Gravity flow is very inexpensive and difficult to control
at flow rates necessary for microscopy. Manual syringes are
ideal for adding growth factors, inhibitors, or other periodic
small-volume fluids. Mechanical pumps are the most reliable
and are available in two popular forms: motorized syringe andperistaltic. The syringe pump is limited in volume for long-term
experiments and subject to flow variations on a micro-flow scaledue to temporary sticking of the plunger. Use caution in selecting
a pump. Stepper motor-driven pumps can be controlled to very
slow flow rates, but the instantaneous rotor movement produceshydrodynamic pulses that can cause the coverslip to flex or thecells to dislodge with the sudden pulse. Before deciding which
type is the most appropriate for the application, consider thefollowing variables:• Expected flow rate
• Uniformity of flow rate over various time scales• Range of flow rate and total volume of flow
• Quality of flow
One pump that has excellent flow characteristics for microscopy
especially at slow flow rates is the model 720 Micro-Perfusion
Pump (INSTECH). It is a peristaltic pump small enough to beheld in the palm of the hand. It uses a tachometer regulated DC
motor and a multistage, step-down gear box to drive the roller
spindle, resulting in a flow profile free from sudden pulsations
typical of most peristaltic pumps. It is equipped with an internal
speed control. It can also be interfaced to a computer throughan analog interface. The pump can be configured to provide
single-channel perfusion for closed chambers or dual-channel
for continuous self-leveling perfusion in an open chamber. Flowrates are adjustable from 2 to 180 ml/hour.

References

DeBiasio R., G.R. Bright, L.A. Ernst, A.S. Waggoner, and
D.L. Taylor. 1987. Five parameter fluorescence imaging: Woundhealing of living Swiss 3T3 cells. J. Cell Biol. 105: 1613-1622.
Focht D.C., Latimer J.J. 2001. Cellular Micro-Observation in
Pharmaceutical Research. Pharmacuetical Visions, Autumn2001:
18-22.
Focht D. C. Farkas D.L. Mammalian Live-Cell Microscopy
Environmental Control. Cell Vision, Journal of Analytical
Morphology. 1995,Vol 2 No.6 450-454.
Farkas D. L., T.B. Ballou, G.W. Fisher, and D.L. Taylor. 1995.
From in vitro to in vivo by dynamic multi-wavelength imaging,
SPIE Prog. Biomed. Optics 2386: 138-149.
Farkas D.L., G. Baxter, R.R. DeBiasio, A. Gough, M.A.
Nederlof, D. Pane, J. Pane, D.R. Patek, K.W. Ryan, and D.L.
Taylor. 1993. Multimode light microscopy and the dynamics ofmolecules, cells, and tissues. Annu. Rev. of Physiol. 55: 785-817.
Inoue, S. 1986 Video Microscopy. Plenum Press, New York.
Pentz S. and H. Schulle. 1981. Revealing the adhesion
mechanism of cultured liver cells to glass surfaces during mitosis
by reflection-contrast microscopy. Zeiss-Inform. Oberkochen 25:
41-43.
Shotton, D.M. 1988. Video-enhanced light microscopy and its
applications in cell biology. J. Cell Sci. 89: 129-150.
Taylor, D.L. and Y.-L Wang. 1989. Fluorescence microscopyof living cells in culture. In Fluorescent analogs, labeling cells
and basic microscopy (part A) and Quantitative Fluorescencemicroscopy-imaging and spectroscopy; (part B). Methods in Cell
Biol. vol. 29 and 30.
Taylor D.L., R. Debiasio, G. LaRocca, D. Pane, P. Post, J.
Colega, K. Giuliano, K. Burton, A. Gough, A. Dow, J. Yu, A.S.
Waggoner, and D.L. Farkas. 1994. Potential of machine-vision
microscopy in toxicologic pathology. Toxicol. Pathology. 22:
145-159.
Webb, W.W. 1986. Light microscopy - A modern renaissance.
Ann. N.Y. Acad. Sci. 483: 387-391.

TubingFor live-cell perfusion experiments, it is prudent to use USPClass VI certified tubing for maximum biocompatibility. There
are three types of tubing that meet this requirement: PharMed(available through Fisher, VWR, and other suppliers), Tygon
2275, and C-Flex. Although PharMed is well suited for cell
culture, it has one main drawback. It is opaque. The perfusate is
not visible through the tubing. Therefore, bubbles that may formin the tubing are hidden from view. This is a major concern whenusing a closed-system chamber because bubbles in a narrow
optical chamber become lodged by surface tension. C-Flex and
Tygon 2275 tubing in l/16 inch size is available nearly clear andnot prone to leaching plasticizers into the perfusate.

9Thermal Reference Images
The thermopgraphic images on these pages represent the propagation of heat in micro-environmental systems. This enables you
to visualize temperature distribution patterns normally not seen by the human eye. These images provide you with a far better
understanding of what is happening on your stage than the limited information on the numerical displays of temperature controllers.
The following thermographic images were acquired at the Bioptechs facility. They were made under laboratory conditions with theuse of a calibrated thermographic camera. An infrared analysis program was used to extract the temperature data and annotate theimages.

Thermal Profile of a Bioptechs Delta T Stage Adapter

Visibile Light Image of a Bioptechs Delta T Stage Adapter

The first Thermograph shows the efficiency, accuracy and uniformity of the Delta T system. Notice the temperature of the stage adapter.
It is nearly the same temperature as the room temperature background. Only the specimen and media are heated. Power consumption
is 0.9 watts because heat is only applied to the specimen area. There is no heat transmitted to the stage. Therefore, it remains “Z” axis
stabile. This is a sharp contrast to traditional peripheral heating methods (shown below), and clearly superior.

Thermal Profile of a Traditional Stage Heater

Thermal Profile of a Traditional Closed Chamber System

This thermograph indicates the disadvantage of peripheral heating.
This is a thermal model of a 50mm culture dish in the center of a
100mm diameter uniformly heated, 3mm thick, aluminum platewith a 25 mm hole in the center. This image was acquired after 20
minutes of equilibration. Note the high temperatures of nearly 60
C, that it takes to reach 37 C in the specimen area. In this case heatthat is not beneficial to the specimen is sunk into the stage causing
Z-axis instability, not to mention the non-uniform temperature of
the specimen area.

This thermograph shows a traditional geometry for a closed
system chamber on a peripherally heated plate equilibrated for 30
minutes. There are two 40mm coverslips separated by a 0.5mm
gasket sandwiched between a 100 mm heated aluminum plate andan acrylic ring cover. Note the temperature of the heated plate
required to warm the center of the field. There is a two-degree
difference between the center of field and the acrylic plate, a ninedegree difference between the heated plate and the specimen area.
This plate is resting in free air, not in contact with a stage plate thatwould sink much of the heat away through conductivity. This excess
heat transmitted to the microscope results in “Z” axis variations.
Also, due to the poor thermal conductivity you can see that it would
take a long time to re-equilibrate after perfusion.

10

Thermal Reference Images

Thermal Profile of a Bioptechs FCS2

Visibile Light Image of a Bioptechs FCS2

The thermograph above demonstrates the uniform temperature distribution of an FCS2. Notice that the coverslip temperature is so
uniform that its location, in infrared, is indistinguishable from the base of the chamber. This demonstrates the effectiveness of the ITO
heated Microaqueduct slide. It is capable of re-equilibrating cell temperature within seconds of perfusion and eliminates the typicalthermal gradient that occurs with peripheral heating.

Thermal Profile of a Bioptechs Objective Heater

Visibile Light Image of a Bioptechs Objective Heater

Notice the temperature distribution in the following locations: nosepiece, bottom of objective, region above heater band, and top of
objective. Power consumption after equilibration is 1.3 watts. The point is that, unless you warm the entire microscope, the microscope
will always act as a heat sink with respect to warming the objective. You can expect a small thermal gradient. All objectives havedifferent thermal profiles. Therefore, it is imperative to efficiently transfer heat to the core of the objective and prevent excess heat fromradiating from the heater-band. This is exactly what the Bioptechs Objective Heater does! Check with Bioptechs for thermal information
on your objective.

11

Bioptechs Stable “Z” Specimen Warming System
Not everyone needs the sophistication of a high-end Delta T Dish System, or the expense that comes with it.
Therefore, Bioptechs has developed a very competitively priced specimen warming system that accommodates any brand of 35mm dish and provides superior thermal control with the exclusive feature of “Z” axis
stability for time-lapse applications. Examine the construction of the Stable “Z” and see for yourself how“stage warmers” should have been made from the beginning! If you need a specimen warmer or if you arestill using a traditional “heated metal plate” to warm your specimen, contact Bioptechs for a demo of this newtechnology!Introducing the Bioptechs Stable “Z” Specimen Warming System
Thermal expansion adjustment ringcompensates for dish height

Adapter plate has a low thermal conductivity and near zero thermal expansion. This
nearly eliminates heat transfer to the stage

Specimen surface iscoplanar with thesupport surface sothermal expansion
does not affect the
specimen plane

Only the specimen retainer
is heated making the mostefficient use of available energy
Ceramic support surface• Stage Geometry to Fit All Popular Microscopes• Adjustable Compensation for Variations in Plastic Ware
• Small Footprint Controller; Easily Mounted on Scope• Powered by 12 V AC Adapter or 12V Battery• Can Accept Any Brand 35mm Dish

• Ambient to 50°C
• Only 4 Watts• Includes Reference Sensor• CE, UL, CSA, Certified• Electrically Quiet

NEW!!!
Now available multi-well heater for Nunc Lab-Tek slides!
Magnetically held pressure piecethat both holds the slide frommoving during stage translationand transfers heat to surroundthe wells
Olympus Stage Adapter

Nunc Lab-Tek 1,2,4 and 8 wellcoverslip bottomed chamber slides
TM

Z-axis stable heating plate
Ceramic support
surface designed to
fit into an SBS plategeometry
Adapter plate has low thermal condutivity and nearzero thermal expansion this effectively eliminates
heat transfer to the stage

Stable Z Controller
12Introducing the Bioptechs Educational (EDU) Series Culture
Dish Control System!
The Delta T EDU is a simplified version of the popular Delta T system that is typically used at the Ph.D. level.
It is designed to enhance the instruction of biology by providing a standardized platform along with a setof instructions that make curriculum relevant demonstrations easy to do and provide a cost savings to theeducational institution. The Delta T EDU gives students the opportunity to experience some of the excitement
or wow factor of observing events that occur in Insect, Yeast, Mammalian, and Avian specimens at the cellularlevel. Some of the demonstrations that can be accomplished in one lab period are: yeast budding, bacterial
invasion and replication, chemically induced reactions of cells, cell to cell interaction, chick embryo heart
rate variations, etc. Bioptechs is currently looking to partner with educators to develop additional live celldemonstration protocols.
Contact Bioptechs if you would like to try a system or suggest a demonstration that you would like to be able
to do in your classroom!

EDU Series Controller

EDU Stage Adapter

Thermal Slide Carrier

Introducing the Bioptechs Thermal Slide Carrier
The Thermal Slide Carrier provides the user with a simple, inexpensive yet accurate means of warming
specimens on a microscope slide on an upright microscope. Simply slide the Bioptechs ITO coated slide into
the carrier and place the carrier into any specimen holder of an upright microscope. A closed loop feedback
controller reads the temperature of the upper surface of the glass slide and applies the appropriate current
to the ITO coated underside of the slide to control temperature. Great for short term observation of live cells.
Compatible with all modes of microscopy!

Revision 5 Bioptechs Controllers!
Bioptechs is pleased to introduce a new generation of temperature controllers with new features!
• Improved analog performance with full digital interface• Greater ease of use, convenient desktop design• Smaller footprint uses less desk space• New GREEN design uses less power and operates more efficiently• Operates from any 12VDC source

Delta T Rev 5 Controller

FCS2 & 3 Rev 5 Controller

13

Objective Heater Rev 5 Controller

Bioptechs Open Dish Systems
An open dish should be used when you need to have physical access to cells
or tissue for such applications as microinjection, electrostimulation, wounding,micromanipulation, use of mechanical probes, recording, and stimulation.
-orAn open system may offer the most economical, convenient, and accurate microobservation method for your application providing an open fluid surface is
acceptable. The following section offers a variety of techniques and supportive
equipment beneficial for the micro-observation of:• Adherent cells• Tissue cultures on a lattice• Suspended cells• Natural or artificial membranes

The Bioptechs’ Delta T® Culture Dish System is an open dish live-cell environmental,
optical containment system. It was designed specifically for live-cell microobservation. Every aspect of its design was engineered for the safety of the cells
and convenience of the user.

Delta T® is ideally suited for use on an
inverted microscope even with high
N.A. lenses.

Delta T® compliments dipping lenses or
LWD 10 - 63X lenses used on an
upright microscope.

Stage heaters are a thing of the past. What you really want and need is fast, efficient specimen
temperature control. Stage heating should be the last thing you want to do because it can cause
Z axis drift. Bioptechs introduced and established The Delta T® Culture Dish System with first-surface direct
thermal transfer and dual mode temperature control for imaging applications in 1993. Bioptechs has now integrated numerous customer requests into this current generation of Open Culture Dish Micro-Environmental
Control System, the Delta T®.
An easily applied continual perfusion system can be employed in either open or closed modes making longterm experiments routine and simplified.

14

Bioptechs Delta T® Open Culture Dish System
Finally, a culture dish system specifically designed for live-cell microscopy! Now
you can have accurate temperature control and high numeric aperture compatibility
in a convenient disposable culture dish system compatible with all modes of
microscopy. Never before has there been so many capabilities integrated into a
specimen control system.
• Easily adapted to a variety of specimen types from monolayered adherent cells to tissue preps• Low mass to thermo-regulate as opposed to conventional stage heaters• Plate, incubate, and observe without the need to transfer your cells• Fast thermal recovery after perfusion (within seconds)• Compatible with inverted and upright microscope stands• Coverglass bottom for optimum optical compatibility• No need for warm air blowers or stage heaters• Direct first-surface heating to your cells• No preheating

Developed and manufactured by Bioptechs, Inc. Pat No. 5,552,321
An intelligent feedback loop passes an electrical current through a thin film coating on the underside of theglass substrate on which the cells are grown. Heat is applied directly to the cells without the inefficiencies
associated with peripheral heating by traditional culture dish warmers. The controller features a real-time temperature display and fast learning curve to compensate for cooling due to surface evaporation while responding to temperature changes due to perfusion. There is also an alarmed protection circuit to safeguard the cells
and an internal reference for user adjustable calibration. The standard controller has a temperature range of
ambient to 50°C. Bioptechs exclusively offers opaque culture dishes which eliminates internal reflection of
unwanted ambient light from the background for fluorescence imaging.

New Features Now Standard on all Delta T® Systems:••••

Mode indicator (dynamic or imaging)
Cold start acceleration
Heat shock activation
Remote setpoint port

• TTL interface and footswitch mode activation• Temperature output port for recording• Heated Lid power supply

You will find the Delta T® a reliable and indispensable addition to your microscope.
15

The Bioptechs’ Delta T® Culture Dish System makes
traditional micro-environmental chambers obsolete!
Limitations of Traditional Technique
•
•
•
•
•
•
•
•

Stage heaters are inefficient, slow, and inaccurate
Plastic dishes are poor conductors of heat
Temperature does not recover quickly during or after
perfusion.
Plastic dishes are not suitable for high resolution or polarization microscopy
Nonuniform temperature distribution
Unnecessary dead volume
Usable aperture of dish limited by the opening in heattransfer plate
Surface evaporation significantly contributes to non-linearity of temperature distribution

Advantages of the Delta T® Dish System
•
•
•
•
•
•
•
•
•
•
•

Place cells onto coverglass and observe
Highly accurate temperature control
Fast thermal recovery
Superior optical image
Stage adapters to fit most popular microscopes
Designed for inverted microscopes but ideal for water
immersion objectives on uprights
Immediate alarm if cell temperature changes
Rigid mount for X, Y stability
Uniform temperature distribution
Cells unaffected by surface evaporation
Numerous specimen adapters available

Originally designed for Inverted
microscopes. However, whencombined with water “dipping”
objectives the Delta T is ideal on
upright microscopes.

•••

Optional heated cover to prevent condensation from forming when used with transmitted modes of microscopy. Bioptechs PN 04200803
For low N.A., use a temperature controller, stage adapter, and a quantity of dishes are all you will need.
However, for high N.A. compatibility, an Objective Heater and an Objective Temperature Controller mustalso be used to assure uniform temperature across the field.
The Delta T controller operates from 100-240 volt AC, 50-60 Hz, or battery. Thermo-regulation is achieved
through control of both amplitude and duration of the current flowing through the ITO coating; average
power to dish is 0.9 Watt.

Bioptechs also offers the Delta T® Dishes without the Indium Tin-Oxide coating for applications where
temperature control is not required. These dishes are available upon special request at a much lower
cost and are compatible with all of the accessories of the standard Delta T® Dish.
16

Delta T® Controller Included Features
Many of these features are made possible due to its fast thermal response which is
not possible with conventional peripheral / radiant heating systems.
Revision 5 Controller

Visual Alarm Led (Red)Colored Display Mode Selection
Red = Setpoint
Amber = Stage
Green = Setpoint
Orange = Heated LidBlue = Imaging
Red = Heat ShockAlarm Reset Button

Power On / Off Button

TTL Port Communicates withAcquisition Host Automatically
Changing Operating Modes
from Dynamic to Imaging andBack During Imaging

Controller Power PlugRemote Setpoint Input V = Temp/10
Analog Temperature Output V = Temp/10
Heated Lid Power Supply

Options for the Delta T® Culture Dish System
Hinged Perfusion Adapter
The Bioptechs Hinged Perfusion Adapter provides Delta T® Culture
Dish users with a convenient and inexpensive method of supporting perfusion needles in the culture dish. The typical application is
to maintain low volume perfusion over cells during long-term experiments. Perfusion adapters are sold in pairs; one hinge and needle isused to irrigate, the other is used to aspirate. The balance between
irrigating and aspirating can be maintained continuously with the useof the Delta T® Micro-Perfusion pump*. Additional supports can be
added to hold gas jets, pH probes, cooling apparatus, or other items
which do not require critical positioning.

Benefits• Needles compatible with 1/16" tubing• User adjustable friction for reliable positioning• Repeatable positioning, flips out when replacing dishes• Adjustable pick-up tube to control level of media in dish
• Perfusion assembly translates with the dish & stage adapter• Eliminates the need for expensive micro-manipulators for
low precision positioning* Bioptechs recommends the use of the Micro-Perfusion Pump withthe double tubing set for closely regulated micro-perfusion.
17

Bioptechs PN 0420081601

Options Continued
Perfusable

Non-perfusable

Bioptechs PN 0420031216

Bioptechs PN 04200312

Coverglass Lid
The Coverglass Lid is a cover for the
Delta T® Culture Dish to be used when imaging to create an optical surface onto theliquid in the dish. This eliminates the optical effect of fluid motion at the air to liquidsurface above the cells that causes thecontrast of the image to change. Therefore, when acquiring a series of images
in a transmitted light, contrast enhancing
mode of microscopy all images will have auniform contrast.

Forming an optically flattened glass-to-media
surface on the top of the cells eliminates thisproblem. The Coverglass Lid fits loosely on
the Delta T® Culture Dish and supports a 1 mm
x 22 mm coverglass in the center of the field
3mm above the specimen. The Coverglass Lid
is reusable and helps the Delta T® bridge thegap between an open dish and a closed system environment. The Coverglass Lid is madeof 304 stainless steel, autoclaveable, andavailable with or without perfusion. This item
and/or the Heating Culture Dish Cover are nearly essential for multi-user facilities.

304 Stainless top
Glass window in contact with themedia provides a uniform andconsistent optical interface for
imaging and/or illumination
Optional 14ga perfusion tubes
1.9mm distance from optical
window to dish surface
Delta T coverglass bottom dishHeating Culture Dish Cover
The Bioptechs Heating Culture Dish Cover is a device, which
provides a condensate-free optical surface above the media
in a Delta T® Culture Dish through which specimens can betransilluminated on an inverted microscope. It is autoclaveable and powered by the Delta T® Controller. The Heated Lidalso provides a gas port to enable containment and control of
atmospheric conditions under the lid.

Bioptechs PN 04200803

The Bioptechs Heated Lid with Perfusion is a device, which
will provide a condensate free optical surface on the top of a
Delta T® Dish through which specimens can be perfused andtrans-illuminated on an inverted microscope. Specimens canbe perfused by attaching perfusion tubing to the ports provided. It is recommended to use the Bioptechs Micro-Perfusion
Pump with the dual perfusion tubing for this purpose.

Bioptechs PN 0420080316

The Heating Culture Dish Cover is made of 316L stainless steel and glass with gold contacts. It is reusable,
autoclaveable, and an ideal accessory for multi-user facilities.
18

Options Continued
Delta T® Cooling Ring
The Bioptechs Cooling Ring is an immersion device
that absorbs heat from the specimen by providing athermally conductive physical barrier between chilled
fluid passing through the ring and the fluid surrounding the specimen. This Cooling Ring is made of
autoclavable 304 stainless steel and provides the microscopist with a convenient and inexpensive methodof reducing the temperature of specimens in Delta T®
Culture Dishes. The Cooling Ring is supported on thestage adapter and translates along with the dish. It iseasily flipped out of the way to enable easy exchangeof dishes in the stage adapter. It is ideal for temporarily cooling the specimen during micro-injection.
Bioptechs PN 04200318
Optional 14ga Perfusion Tubes
Stainless Steel Adjustment Ring
Optical Glass Surface

Media Volume Reducer
The Media Depth Reducer allows for the reduction of media
volume above cells adherent to the coverslip bottom of a
Delta T Dish. Reducing this volume may be desirable when,for instance, background fluorescence is a problem, or when
the cost of a reagent dictates the use of small volumes.
The Media Depth Reducer is composed of a 1mm thickcoverglass window mounted in an externally threaded black
Radel insert, which in turn mates to an internally threaded
metal mounting ring. The height of the coverglass window
relative to the coverglass bottom is continuously adjustableby rotating the Radel insert.
Bioptechs PN 04200303

Delta T Coverglass Bottom Dish
Depth Adjustable by Rotation of Adjustment Ring

Yeast Slide
The Bioptechs Yeast Slide provides an opticallytransparent, temperature controlled surface on
which to place specimens on an upright microscope. It is a glass slide with an ITO coating on
its bottom surface measuring 50mm X 75mm. It
is placed on the stage of an upright microscopeand mechanically constrained by conventional
means. It accommodates a standard 25 x 75mm
slide having yeast or other specimens under acoverslip. Its temperature is maintained by anelectronic controller that reads temperature fromthe slides integral thermal sensor and heats bypassing a current through the ITO coating on theunderside of the slide.
Bioptechs PN 042005076

19

Tissue Slice or Artificial Membrane MicroObservation Utilizing Delta T® Culture Dish System
The Bioptechs Delta T® Culture Dish System gives you a great deal of flexibility never before achievable withtraditional culture dishes. To observe a specimen, you only have to place it in the Delta T dish within the focal
plane of the objective and let the Delta T® system maintain the temperature, optical, and fluid requirements.
Bioptechs manufactures several adapters to provide a convenient method of micro-observation of live tissue or cells grown on artificial membrane. Refer to the diagrams below for samples of the devices currently
available or consider the advantages of combining one of the traditional inserts available from Costar, Falcon,
Nunc, etc. with the Delta T® Culture Dish System.
Specimens are defined onto a plane parallel to the coverslip for flat-fieldobservation and suspended in the dish by a Radel holder threaded into an
adjustment ring.
Both the upper and lower contact surfaces of the dish are precision
injection molded to be used as optical reference planes.

Tissue Slice Adapter for Delta T Culture Dish System
Stainless Steel Adjustment Ring
Tissue Slice Adapter is Adjustable in
HeightOptional 14ga Perfusion Tubes
Delta T Coverglass Bottom Dish
Ruber O-Ring to Secure Membrane in Place
Tissue Slice Membrane
Depth Adjustable by Rotation of Adjustment
Ring

Bioptechs PN 042021918

Membrane Insert Adapter for Delta T Dishes
The Delta T environment is easily adapted to accommodate manyof the artificial membrane culturing products on the market. The
device shown supports a Costar Transwell membrane. The bottom
surface of the membrane is observable on an inverted microscope
and adjustable in the “Z” axis to enable accommodation of higher
numeric aperture lenses. Perfusion ports can be used to perfuse
the basil surface of the specimen. Both surfaces of the membrane
can be perfused separately.
•
•
•
•
•

Tubing connections 1/16”
Fits only Delta T Dishes
Reusable
Autoclaveable
When ordering this device, Bioptechs requires a sample of theinsert to be used.

Bioptechs PN 0420011301

20

Brain Slice Micro-Observation Chamber
for Inverted Microscopy
The Bioptechs Brain Slice Adapter is combined with the Delta T® Culture Dish System to provide a convenientmethod of observing thick cut sections of brain or other tissue in a perfusable, temperature controlled, opticalenvironment on an inverted microscope. Perfusion ports are made of 304 stainless steel and are compatible
with 1/16" tubing. As with all Bioptechs Delta T® Culture Dish Adapters, the specimen is adjustable in the Z
axis plane to accommodate the working distance of the objective. Custom geometry adapters are available
upon special order to accommodate specimens having unique geometry.
Radially Porforated for Media Exchange
Nylon Mesh-Bottomed Weight

Brain Slice Insert
Stainless Steel Adjustment Ring
Optinal 14ga Perfusion Tubes
Delta T Dish
10mm Dia x 0.5mm Polycarbonate Disk with
1.5mm Aperature in center
The Specimen Rests on the Aperature while
Supported in the Delta T Environment

Bioptechs PN 0420201919

Depth Adjustable by Rotation of Adjustment Ring

•
•
•
•
•
•

The specimen is placed onto a polycarbonate disk with an aperture in its center
This disk is placed into a well / receiver in the Z adjustable support
A 304 stainless steel weight with a Nylon mesh covering rests on the specimen to keep it from floatingThis assembly is threaded into a focusing ring that rests on the top of the Delta T Dish
The depth of the specimen in the dish is adjusted by rotating the focusing ring
Perfusion occurs around all surfaces of the specimen

Atmospheric Barrier Ring For Water “Dipping” Objectives
on Upright Microscopes

The Atmospheric Barrier Ring is made ofborosilicate glass, autoclaveable and available inseveral sizes to fit most popular water “dipping”
lenses. It is placed around the barrel of water“dipping” lenses on upright microscopes toreduce ambient contamination and evaporation,
thereby increasing humidity above the specimen
and helping to retain the pH during long-termtime lapse imaging.

Bioptechs PN 04200325

21

Automated Perfusion Configuration for
Delta T™ Culture Dish System

The Delta T Micro-Perfusion Pump is equipped with a dual tube manifold. The tube with a black ring on it hasa slower fluid transfer rate than the other tube. The fresh media supply is coupled to this slow flow tube andthe dish aspiration needle is attached to the faster flow tube. The level in the dish is controlled by adjusting
the depth of the aspiration needle. This technique works well with all Delta T perfusable accessories.
Note: The Hinged Perfusion Adapters (shown) can be replaced by the Perfusable Coverglass Lid for closed
system perfusion.
22

23

Bioptechs Delta T System Media Perfusion Diagram with CO2 Regulated MediaBioptechs Delta T System with CO2 Perfusion Above the Media Diagram

24

Bioptechs Delta T® Stage adapters
The Bioptechs ∆T® Stage Adapter reads the temperature of ∆T Culture Dish, provides electrical contacts
to power the dish, and supports the dish on the stage for translation. All ∆T systems require aStage Adapter. These images are provided to assist in identifying and selecting a stage adapter appropriate
for your microscope.

Single plate stage adapter fits:
Zeiss Axioverts
Leica DMIRB
Bioptechs PN 04202601

Leica DMIRB triple plate stage adapter
Bioptechs PN 04201203

Triple plate stage adapter fits:
Axiovert, Ludl, Prior, and ASIstagesBioptechs PN 04202602

Round stage adapter fits: Nikon, Olympus,
and Delta Vision stage openings
Bioptechs PN 04201415
To fit Zeiss Gliding stage PN 04202603

Multi-well plate size adapter fits
into any multi-well receiver
Bioptechs PN 04202003
Cell Robotics size PN 04201220

Delta T Upright stage adapter fits
most upright microscopes
Bioptechs PN 04202116

Leica Galvanic rectangular stage insert
Bioptechs PN 04201807
Marzhauser Galvanic rectangular stage insert
Bioptechs PN 04202807

Leica Galvanic round stage insert
Bioptechs PN 04201207
25

Accessories
Bioptechs Culturing Cylinders:
Culturing Cylinders are used to barricade cells or suspended specimens in a Delta T® Culture Dish or to restrict and concentrate the growth and location of cells
plated on an FCS2® Coverslip. They are 5mm highand available in a variety of inside diameters including
1mm, 2mm, 4mm, 6mm, 8mm, 10mm, and 12mm. The
outer diameter is always 2mm greater than the inside
diameter due to the 1mm wall thickness. The cylinders
are made of Pyrex glass and are heavier than plasticcloning rings to eliminate floating. They are optically
See Price List for Bioptechs PN
polished on the bottom surface to mate with and forma tight seal with other glass surfaces such as FCS2® Coverslips
and Delta T® Culture Dishes without the need of grease. Culturing Cylinders can be autoclaved for reuse.
No grease, no contamination, reuseable!

Delta T® Culture Dish Application:
When using Culturing Cylinders with the Delta T® Culture Dish, select the appropriate size cylinder for the protocol and place the cylinder into the center of the dish.
Pipette the specimen into the cylinder. Then pipette media around the cylinder to adepth equal to the depth in the cylinder, thereby equalizing hydrostatic forces.

FCS2® Application:
Place the appropriate Culture Cylinder onto the center of the 40mm FCS2® Coverslip to restrict plating of the cells to the central region of the Coverslip. Apply
media around the cylinder to equalize hydrostatic forces. After cells have grown toconfluence, remove the Culture Cylinder and transfer the coverslip to the FCS2®
for micro-observation.

Boekel Desktop Warmer
When working with live cells on a microscope, it is convenient
to have an incubator close to the scope. It is beneficial for
temporary storage of chambers as well as specimens or media to keep them prewarmed. Most importantly it is a clean,dry, and safe place for objectives when not being used. Bioptechs recommends the Boekel Warmer because of its small
size and light weight. It is not humidified, making it an ideal
warmed objective storage device. The Warmer measures 8”
x 8” x 7”, operates on 120V AC, and is small enough to be
mounted close to the microscope for temperature controlledstorage of perfusion media during experiments.
Bioptechs PN 260700

26

Bioptechs Closed Chamber Systems
Closed systems are used for the following applications:• The experiment requires that the specimen be contained in a completely airtight environment.
• The optical constraints of the microscope dictate a more precise definition of optical surfaces.
• The experiment requires discrete control of the flow characteristics (shear-force issues).
Bioptechs offers an advanced common sense approach to live-cell environmental control. The
FCS2® Closed System along with the Objective Heater® for high numeric aperture applications is the mostreliable and easily modifiable method for time-lapse micro-observation of living cells available.
Eliminates the need for:• Warm air curtains (blow dryers)• Peripheral stage warmers• Heat lamps• Microscope enclosures

The following section offers an integrated solution for closed-system, micro-environmental control completewith near laminar perfusion, user definable volume and flow, and optical compatibility with all modes of lightmicroscopy.

Eight-hour time lapse of normal mammary epithelial cells at 250X DIC. Mammosphere formation is visible inthis video. (The round cluster of cells in the center of the field is a mammosphere.)Compliments of Jean Latimer Ph.D. Magee Womens Research Institute, Pittsburgh, PA.

Tissue: Human Breast Tumor Cells
Total acquisition Time: over 5 hours (1 frame every 10 minutes)
8 pane image represents 1 frame every 30 minutes
200X DIC Zeiss Axiovert, QED Imaging, Bioptechs FCS2® Chamber System
Image compliments of Jean Latimer, Ph.D., Magee Womens Research Institute, Pittsburgh, PA
27

In order to accurately record the morphology of your cells, youwill need a micro observation environment that is both conduciveto their viability and compatible with all techniques of microscopy!
The Bioptechs’ Focht Chamber System (FCS2®) is a state-of-the-art, parallel plateflow cell with uniform temperature control and a user definable flow cavity that iscompatibile with all modes of light microscopy including: BF, DF, PH, DIC, Hoffman,
Fluorescence, Confocal, and Multi Photon.

Developed and manufactured by Bioptechs, Inc. patent #s 4974952, 5414556 and 5410429• Microaqueduct design enables proper Koehler illumination with high numeric aperture optics for bothtransmitted and reflected modes of microscopy• Suitable for no flow through high flow rate procedures where a rapid exchange of media is required
and discrete control of cell surface shear is user definable• Cell temperature can be controlled from ambient to 50°C ± 0.1°C without the need of an air curtain• Temperature is controlled uniformly across entire field with media equilibrating as it enters the chamber• Compatible with 1/16" tubing for perfusion• Easily assembled with ordinary skill (no tools required)• Stand-alone temperature controller with an alarm interrupt circuit to safeguard your cells• Objective Heater® and Objective Temperature Controller required for high N.A. applications• Remote setpoint interface and/or chart recorder output now included on all systems•

Closed system so that CO2 dependent media can be employed28

Bioptechs FCS2® Technical Data
The patented FCS2® Microaqueduct Slide technology, with its heated optical thin film coating, provides highnumeric aperture compatibility, temperature control without an air curtain, high volume perfusion rates, user
definable volume, and control of cell surface shear. The optical design of the FCS2® is based on slide andcoverslip geometry to provide compatibility with all microscopy techniques.

Specifications:Physical size
Coverslip #1.5 thick X
Imaging aperture
Maximum volume
Minimum volume

75mm OD, 13mm high40mm Diameter
22mm
0.706µL
0.043µL

Maximum volume exchange rate
Minimum fluid aperture
Separation between optical surfaces
External port ID
Temperature stability

The controller features separate channels for the
microaqueduct slide and metal chamber base plus:• Large backlit LCD temperature display• Logic circuit safeguards cells• Built in calibration• Audible and visible LED alarm• Chart recorder output for temperature recording• Remote setpoint for temperature programming• Auto reset• Dual voltage operation 120 - 240V AC 50 - 60 Hz

1/sec
0.63mm^2
50 - 1000µ1.6mm±0.1°C

Bioptechs PN 060319-2-03 (FCS2 Chamber)

Temperature Sensors
Heater Contacts
Perfusion Connections

The volume of the chamber is determined by both the
thickness and internal geometry of thelower gasket. The flow
characteristics can befurther modified bychanging the internal geometry of thisgasket. An assortmentof gaskets shapes andthicknesses as well as
blank gasket material is included in thestarter kit.

Upper Gasket

Microaqueduct Slide (opticalsurface, perfusion guide andtemperature control)

Lower Gasket
Defines Flow Geometry

40 mm, #1.5
Coverglass

The FCS2 Starter Kit Includes
Chamber, Controller, 5 Microaqueduct slides, 30 piecegasket set, 50 - #1.5 coverslips. The Objective Heaterand Objective Heater Controller is optional (needed foruse of high numeric aperture objectives).
The objective heater is mounted to the upper end of
the central retracting tube of the objective. The standard size fits diameters between 17 and 28mm. The
medium size fits diameters between 23 and 29mm.
There is also a large size to fit objectives with 26 35mm diameter.

Self-Locking
Base with
Internal
Temperature
Control

Objective Heaterfor High N.A. Lenses29

30
6 Luer adapter (male, adapts 1/16" tubing to female Luer lock)7 Alternate media glass syringe (growth factors, inhibitors, etc.)
8 Three-way valve (switches source of perfusion; pump or manual)
9 FCS2 Chamber
10 Tubing to waste container
11 Tee adapter (place at same height as chamber, open top of tee to prevent siphoning)

Note: Syringes are glass to prevent contamination of media from silicone lubricant typical in plastic syringes.

1 Perfusion tubing, 1/16" ID supply line (keep as short as possible)
2 FCS2 Micro-Perfusion Pump (recommended for prolonged, controlled
perfusion in delicate optical cavities)
3 Tee adapter (adds low volume syringe to facilitate manual perfusion whenloading chamber)
4 Luer adapter (female, adapts Luer taper syringe to 1/16" tubing)
5 Low volume glass syringe

Typical FCS2® Perfusion Configuration for Induced Change Experiments

31

CO2 Enriched Media Schematic for Bioptechs FCS2 & FCS3 systems.
Easy to setup, reliable and consumes very little CO2

Bioptechs Options for the FCS2®
Open Mode Top for the FCS2®
The open mode option allows the FCS2® to be assembled
without the microaqueduct slide thus exposing the cells on
the coverslip for microinjection. The coverslip can then be
removed and reassembled with the microaqueduct slide forlong term time-lapse applications.

Open Mode Adapter

Bioptechs PN 060319-2-1513

Installed on FCS2 Base

Custom Microaqueduct Slides
The standard Microaqueduct Slide is made withtwo “T” shaped grooves 22mm apart in a pattern
optimized for uniform perfusion. If an experimentrequires a narrow flow path or low dead volume,
Bioptechs can custom machine the microaqueduct slide to your specifications.
Bioptechs PN 130119-5

Call for Bioptechs PN

Custom Gaskets
The standard gasket set included in the FCS2® Starter Kit contains gaskets of various thickness (ranging from100 to 1000 microns) and internal geometry. Included are blank gaskets that can be customized by the enduser. However, in some cases an end-user may want a supply of a special shape or thickness of gasket. The
Bioptechs Custom Design Department can make these special items. We require a one-time setup charge inorder to make any number of gaskets an end-user wants at the standard gasket price. Standard Gasket Set
Includes: (3) 0.1mm 30mm Round, (2) 0.1mm 14 x 24, (3) 0.25mm 30 mm Round, (2) 0.25mm 14 x 24, (1)0.25mm Blank, (3) 0.5mm 30mm Round, (2) 0.5mm 14 x 24, (2) 0.5mm Blank,(5) 0.75mm 30mm Round w/
holes, (2) 0.75mm 14 x 24, (1) 0.75mm Blank, (2) 1.0mm 30mm Round, (1) 1.0mm 14 x 24, (1) 1.0mm Blank

30mm Standard
Round

14mm x 24mm
Rectangle

30mm Round w/
perfusion holes

Blank 40mm

Tubing Adapters
Connecting tubing to syringes and chambers may require
special adapters. Bioptechs has assembled a kit of connectors and adapters which conform to the specificationsin the FCS2® perfusion drawing (previous page). The kit
components are pictured.
Bioptechs PN 162003-1

32

Closed-System Cooled Chamber
FCS2® Cooling

Refrigerant Fluid PortFCS2 Cooling Adapter

FCS2 Cooling Adapter

This configuration is designed to allow high
N.A. observation of specimens at below
ambient temperatures on an inverted
microscope. This design provides the same
optical and flow characteristics as thewarmed FCS2
but provides an adjacent secondary perfusion chamber through which a refrigerant
fluid is circulated. Caution: When working
below ambient temperatures with highnumeric aperture lenses, an Objective
Cooling Ring and Objective Thermal Isolator should be used.

FCS2
Top

Exploded View

FCS2
Base

Note: optional tubing can be installed through which refrigerant fluid can be circulated. The coolant connection tubes exit the FCS2® from above. As shown in the picture below.

Assembled Cooled FCS2 With Cooling Adapter
Bioptechs PN 03060319-2

FCS2 with Cooling Adapter installed
Note: auxiliary tubing on the adapter
Bioptechs PN 03060319-2(Cooilng Adapter Only: 060319-2-0301)

How it works:
Cells are plated onto a 40 mm coverslip. Then the coverslip is placed into a modified FCS2® chamber. This
provides a perfuseable laminar flow optical chamber with user modifiable flow characteristics. The upper glass
element (microaqueduct slide) is then used to remove heat from the specimen cavity to be absorbed by thecooled fluid being circulated in the second, parallel cavity formed by the addition of an o-ring sealed window
adapter. Gravity or an electric pump is then used to create a flow of chilled fluid through the heat exchange
cavity in the chamber. The cells remain safely enclosed in a separate optical enclosure. If more thermal transfer is necessary, tubing can be installed into the base of the FCS2®.
Note: If you are using high N.A. objectives at below ambient temperatures, it will be necessary to cool andthermally isolate the objective from the microscope. The optional thermal isolator will prevent condensationfrom forming on the lower element of the objective. Four gas ports are provided to purge the adapter with dry
air. An anti-reflection coated window optimized for 340-700 nm seals the bottom of the adapter. See page 40.

33

Bioptechs FCS2® Stage Adapter Identification
The FCS2® Starter Set requires a stage adapter for precise positioning. Due to the diversity of microscopestages, a stage adapter cannot be included with the FCS2® Starter Set. Therefore, you need to select a stage
adapter for your scope from the illustrations below. When ordering, please indicate the brand of microscope,
the manufacturer’s stage identification number, and description, along with the corresponding Bioptechs’ partnumber. Custom designs are available upon special order.

Round stage adapter fits:
Olympus IMT2, IX70. IX50
Nikon Diaphot, TE200-300
and Deltavision
Bioptechs PN 060319-2-1415

Triple plate stage adapter fits only:
Leica DMIRB
Bioptechs PN 060319-2-12

Triple plate stage adapter fits:
Zeiss Axiovert, Ludl, Prior, and ASI Stages
Bioptechs PN 060319-2-2611

Single plate stage adapter fits:
Zeiss Axiovertl, Leica DMIRB and DMIL
Bioptechs PN 060319-2-2613

34

Bioptechs Upright Microscope Compatible
Closed Chamber System (FCS3®)
The Bioptechs’ Focht Chamber System 3(FCS3®) is basically a redesign of the popular FCS2 but optimized
for use on an upright microscope. The FCS3® uses the same, microaqueduct slide, gaskets, and, coverslips
as the inverted model. Therefore, it has all of the same operating characteristics as its predecessor. The
main difference, in the upright version, is that the coverslip portion of the optical cavity is oriented upward
toward the objective*. As an added benefit, the upright version uses a universal stage adapter to fit all uprightmicroscopes. By simply replacing the specimen carrier with the upright FCS3® interface plate, any uprightmicroscope can become a multi-functional live-cell workstation.
The Upright FCS3® provides all the functionality of the inverted model (FCS2®) including:• Microaqueduct design enables proper Koehler illumination with high numeric aperture optics for both transmitted and reflected modes of microscopy• Suitable for no flow, through high flow rate procedures where a rapid exchange of media is required anddiscrete control of cell surface shear is adjustable• Cell temperature can be controlled from ambient to 50°C ± 0.1°C without the need for an air curtain• Temperature is controlled uniformly across entire field with media equilibrating as it enters the chamber• Closed system so that CO2 dependent media can be employed• Compatible with 1/16” tubing for perfusion• Easily assembled with ordinary skill (no tools required)• A stand-alone temperature controller with an audible alarm and interrupt circuit to safeguard your cells• Objective Heater and Objective Temperature Controller required for high numeric aperture applications• Includes the capibility to interface to a computer for temperature programming and/or chart recorder output• Fits most Leica, Nikon, Olympus & Zeiss upright microscopes (custom stage adapaters available)

FCS3 & Universal Upright Stage Adapter

The optical cavities supportive structure positions the microaqueduct slide 5mm millimeters above the stage
surface with the coverslip facing upwards towards the objective. The additional space between the top surfaceof the stage and bottom of the microaquaduct slide is required for electronic connections and perfusion ports.
It may be necessary, on some scopes, to readjust the condenser stop screw to accommodate this additional
distance. No other modifications should be necessary.
35

Bioptechs FCS3®
Temperature Control:
The FCS3 was designed to maintain accurate thermal control and allow near laminar flow perfusion. Both ofthese functions are incorporated into our patented Microaqueduct Slide (see drawing below). The surface ofthe slide, opposite the specimen side, is coated with an electrically conductive transparent thin film of IndiumTin Oxide (ITO) and two electrical contacts (busbars). When the FCS3 is completely assembled and placedon the stage adapter, two electrical contacts and a thermal sensor, (not shown in drawing), rest on these busbars. A temperature controller is used to pass a regulated current flow through the ITO Coating. This causes
the surface of the slide to heat. The heat is transferred through the media to the cell surface on the coverslip
thereby providing a conductive heat transfer. The metal base of the chamber is also temperature regulated toprovide heat to both the incoming media and peripheral thermal support to the metal housing.
Microaqueduct Perfusion:
A fluid pathway is formed by separating the Microaqueduct slide from the coverslip containing cells with asingle silicone gasket. This gasket can be any thickness from 50 micron to 1mm and any lateral geometry you
choose or create. This arrangement allows the user to define the flow characteristics. Therefore, you are notlimited by the geometry of the optical cavity instead you select or create it! Fluid access to this flow channel
is made through two 14-gauge needle stock tubes protruding from the sides of the chamber top. These tubes
provide fluid connection to two perfusion holes in theMicroaqueduct slide that interface two “T” shaped grooves
cut into the inner surface of the Microaqueduct slide. The“T” groove allows the media to seek the path of least resistance and become nearly laminar before flowing acrossthe cells. This technique eliminates the need for the metal
perfusion ring and additional gaskets, which are the limiting factors, required by most conventional chambers.

Microaqueduct design enables proper Koehler illumination with high-numeric aperture optics for both transmitted and reflected modes of microscopy
•
•
•
•
•
•
•
•

Suitable for no flow through high rate flow procedures where a rapid exchange of media is required withlow cell surface shear
Cell temperature can be controlled from ambient to 50 degrees C +/- 0.2 degrees C without the need of an
air curtain
Temperature is controlled uniformly across entire field with media equilibrating as it enters the chamber
Closed system so that bicarbonate CO2 or organic buffers can be employedCompatible with 1/16” tubing for perfusion (C-Flex, Tygon, etc.)
Easily assembled with ordinary skill (no tools required)Stand-alone temperature controller with an alarm circuit to safeguard your cells
Near laminar flow
36

Bioptechs FCS3® Technical Data
The FCS3® utilizes a patented Microaqueduct Slide technology, having an electrically conductive, opticallytransparent thin film coating, on its outer surface to provide the following attributes:• High numeric aperture compatibility• Temperature control without an air curtain• High or low rate perfusion capability• User definable volume, and control of cell surface shear• Compatibility with all modes of microscopySpecifications:Physical size
Coverslip #1.5 thickImaging aperture
Maximum volume
Minimum volume

75mm OD, 13mm high40mm Diameter
28mm
706µL
43µL

Maximum volume exchange rate
Minimum fluid aperture
Separation between optical surfaces
External port ID
Temperature stability

The controller features separate channels for the
microaqueduct slide and metal chamber base plus:• Separate channels for the microaqueduct slide andchamber base• Large backlit LCD temperature display• Logic circuit safeguards cells• Built in calibration• Audible and visible LED alarm• Chart recorder output for temperature recording• Remote setpoint for temperature programming• Auto reset• Dual voltage operation 120 - 240V AC 50 - 60 Hz*The volume of the chamber is determined by both the

1/sec
0.63mm^2
50 - 1000µ1.6mm±0.1°C

Bioptechs PN 21-060319-3-03 (FCS3 Chamber)

Chamber Top

Pressure Plate

thickness and internal geometry of the upper gasket.
Changing the internal geometry of this gasket canfurther modify the flow characteristics.
40mm Coverslip

Upper Gasket*

Microaqueduct Slide

Lower Gasket
Perfusion Tubes(14 gauge)

FCS3 Cross Sectional View

The FCS3 Starter Kit Includes

Chamber Base

Universal upright stage adapter plate, Chamber,Controller, 5 Microaqueduct slides, a 30 piece gasket
set including an assortment of various shapes andthicknesses as well as blank gasket material for user
customization, and 50 - #1.5 40mm coverslips.

An Objective Heater and Controller are optional (needed for use with high numeric aperture objectives). The
objective heater is mounted to the upper end of the central retracting tube of the objective. The standard size
fits diameters between 17 and 28mm. The medium size fits diameters between 23 and 29mm. There is also alarge size to fit objectives with 26 - 35mm diameter. (See page 43)37

6 Luer adapter (male, adapts 1/16" tubing to female Luer lock)7 Alternate media glass syringe (growth factors, inhibitors, etc.)
8 Three-way valve (switches source of perfusion; pump or manual)
9 FCS3 Chamber
10 Tubing to waste container
11 Tee adapter (place at same height as chamber, open top of tee to prevent siphoning)

Typical FCS3® Perfusion Configuration for Induced Change Experiments

1 Perfusion tubing, 1/16" ID supply line (keep as short as possible)
2 FCS3 Micro-Perfusion Pump (recommended for prolonged, controlled
perfusion in delicate optical cavities)
3 Tee adapter (adds low volume syringe to facilitate manual perfusion whenloading chamber)
4 Luer adapter (female, adapts Luer taper syringe to 1/16" tubing)
5 Low volume glass syringe

Note: Syringes are glass to prevent contamination of media from silicone lubricant typical in plastic syringes.

38

Bioptechs Objective Temperature Control
The Problem:
When high numeric aperture objectives are used to observe temperature sensitive specimens, heat from thespecimen is transferred through the optical coupling medium (oil, glycerine, or water) to the colder objective.
Therefore, it is necessary to control the temperature of the objective. It is important to understand that when thesystem is set up properly, the objective heater is only used to prevent heat loss from the specimen. It should not
be used to provide heat to the specimen.

The Wrong Solution:
Never use an objective heating device that does not transfer heat to the objective efficiently. Any heat that does
not go into the objective will escape, convect upward and “overheat” your specimen!

The Right Solution:
To eliminate the heat loss from the specimen, Bioptechs developed a patented Objective Heater System®, which
includes a heater/sensor and an electronic controller specifically designed for this purpose. The heater/sensor
is comprised of an adjustable thin-film heating band which surrounds 3/4 of the diameter of the upper region of
the central retracting tube of the objective. A temperature probe positioned in the gap formed between the endsof the heating band provides accurate feedback to a closed loop controller. A metal cube shaped frame supportsa thermal sensor and contains a mechanism to adjust the size of the heater-band.
The Objective Controller® is specifically designed to slowly heat the objective over a fifteen minute warm-up
period, then hold the objective at the setpoint value within 0.1°C. The Controller operates from ambient to 50°C
and has special safety circuitry which utilizes a 0.9°C error window to shut down the Controller and sound an
alarm if the temperature of the objective deviates after it has reached setpoint.
The heater loop requires a minimum of 5mm longitudinal physical contact with a cylindrical surface on the objective. In some cases there may be a decorative collar on the objective that must be removed in order to permit
adequate surface contact and efficient transfer of heat.

Special note:
1. When using warmed objectives it is recommended to use Type 37 Immerion Oil, available from Bioptechs.
This oil is specially formulated to have a refractive index of 1.518 at 37°C.
2. The Bioptechs’ Objective Heater® can be adapted to fit all objectives. Due to the size, geometry and thermal characteristics of some objectives, it may be necessary to use a thermal spacer to eliminate the influenceof the nosepiece.
High efficiency thermal transfer,consumes less power and noradiation of excess heat

Band tightening screw on backside for more convenient accessRemovable power cord so theheater can remain on the objective
while it is rotated in the nosepiece

Second retainer loop for remarkably secure attachment,
easily tightend with the pushof a button

Highly flexible power cord

New smaller cube design, fits all
objectives in any orientation
Bioptechs Objective Heater® mounted to a 100X lens with a Thermal Spacer

39

Objective Accessories
Bioptechs Objective Cooling Collar
The Bioptechs Cooling Collar is an attachment device designed toremove heat from an objective through fluid transfer. The Cooling
Collar provides an isolated fluid passageway for chilled fluid to circulate around the objective thus absorbing heat by conduction to theadjacent metal surface. A separate chiller bath and circulating pumpis required.

Bioptechs PN 150303

Objective Thermal Isolator and Spacers
The Bioptechs’ Thermal Isolator is made of a material having
low thermal conductivity for mounting objectives to thenosepiece of a microscope, which provides a gas trap to
enclose the exposed optical elements from condensation. It isnecessary to provide a supply of dry air or gas into the baseof the Thermal Isolator. The Thermal Isolator will add 9mm to
the height of the objective. Therefore, it is usually necessary
to elevate the stage by the same amount. Due to the wide
variety of stages available, Bioptechs does not provide
spacers for stage mounts. When the height of a microscopestage is adjusted by 9mm to accommodate the addition of a
Thermal Spacer to one of the objectives, it is often convenientto adjust the height of the other objectives on the turret by thesame amount. The Objective Spacer functions to add 9mms ofheight to an objective. Note: When ordering Objective Thermal
Isolators or Spacers you must specify the thread type.

• RMS = 0.8" x 36 tpi (approximately 20mm diameter)• 25mm x 0.75 mm/thread (approximately 1" diameter)

Thermal Isolators

Thermal Spacers
For RMS Thread:
Bioptechs PN 152009R - Isolator
Bioptechs PN 152019R - Spacer
For 25mm x 0.75mm Thread:
Bioptechs PN 152009 - Isolator
Bioptechs PN 152019 - Spacer

Bioptechs Objective Wrench & Pliers
In order to expose sufficient surface for mounting heat transfer devices on some manufacturers’ objectives, itis necessary to remove the outer decorative collar which contains written information such as magnification,
N. A., etc. The objective wrench is designed to enable the user to isolate stress applied to an objective whenremoving this collar. The rotational force applied to the objective must be applied exclusively to the mounting threads and the cover. The wrench, designed with two thread sizes (0.8" X 36 TPI (RMS) and 25mm X
0.75mm/thread), securely binds these threads and provides sufficient leverage for removal of the cover.
Objective Wrench & Pliers

Objective with decorative collar removed

Bioptechs PN 021523

40

Micro-Perfusion Pump
When imaging cells on a coverslip in a chamber where liquid is confined, any changes in flow rate can translate to changes in pressure. This causes the coverslip to exhibit diaphragm-like behavior. Given the narrow
depth of field of microscope objectives, this coverslip flexing causes the cells to go out of focus. To eliminate
this problem at the lower flow rates and significantly reduce it at higher flow rates, Bioptechs recommends theMicro-Perfusion Pump for use with all its micro-observation systems.
The Micro-Perfusion Pump is a miniature, single or dual, channel full-featured peristaltic pump designed
specifically for low-flow rates. Unlike most peristaltic pumps that are driven by stepper motors, the Micro-Perfusion Pump is driven by a tachometer regulated, multi-stage DC gear motor. This assures a smooth analog
rotation of the roller spindle, free of instantaneous steps. It is regulated by either the internal control circuitry
adjustable from 2-180 ml/hr or it can be interfaced with a computer through a DIO port. The pump comes withan external 9 Volt AC adapter, and also contains an internal 9 Volt battery which can function as the primary
power supply if needed.
The pump includes a single 0.062" I.D. tube set for use as a single channel pump or a dual tube set that has
two 0.062" I.D. tubes for use as a dual channel-pump. Although other tubing sizes are available, these twosizes are best suited for imaging applications. The pump tubes are made of C-Flex and are terminated witha 1/16” tubing barb. The base of the pump is threaded for easy mounting to a stand or fixture near the microscope.

Features• Compact• Usable as either single or dual channel• Precise speed setting feature• Flow rates of 2 to 180 ml/hr• Compatible with 1/16" tubing• External DC power supply with internal 9 volt backup battery• Can be operated on internal battery if desired• Preassembled autoclavable pump tubes• Computer interfaceable

Back, Internal or External Speed Control,
Accepts External Power Suppy.

Front , Accommodates One or Two Tubes

Bioptechs PN 060319131616 (FCS2)
Bioptechs PN 0420131616 (Delta T)

41

Options for the Micro-Perfusion Pump
Rod Mounting Clamp for MicroPerfusion Pump
The Micro-Perfusions Pump Rod Mounting Clamp
provides a method of mounting perfusion pumps
onto a conventional one-half inch support post.
This makes positioning perfusion pumps adjacentto the microscope easy and convenient.

Bioptechs PN 16181303

Single Channel Tubing Assembly for
Micro-Perfusion Pump
The Single Channel Tubing Assembly for theMicro-Perfusion Pump is a precise length of either
C-Flex or Silicon tubing, 0.0625" ID, bonded totermination blocks that anchor to the pump andprovide 1/16th inch barb ends to attach additional
tubing. The Single Channel tubing is typically used
for FCS2 perfusion.

Bioptechs PN 060319192016 for C-Flex
Bioptechs PN 060319192016S for Silicon

Dual Channel Tubing Assembly for
Micro-Perfusion Pump
The Dual Channel Tubing Assembly for theMicro-Perfusion Pump is made of two precise
lengths of either C-Flex or Silicon Tubing bonded
to termination blocks that anchor to the pump andprovide 1/16th inch barb ends to attach additional
tubing. The dual channel tubing is typically used
for perfusion of the Delta T and its accessories.
Note: The tube with the black band attached has
a slightly smaller ID. Therefore the fluid flow is
reduced through that tube as compared to theunmarked tube. When properly assembled (seeillustration on page 19) this slight disparity in flow
rates enables worry-free control of the fluid level inthe dish.

Bioptechs PN 0420042016 for C-Flex
Bioptechs PN 0420042016S for Silicon

42Perfusion and Temperature Control For Live-Cell Imaging!
The live-cell microscopy Perfusion and Temperature Control Interface from
Bioptechs™ provides the following valuable features:
Extreme ease of use (fill in the blanks) for precise and repeatable control of:• Dual or single micro-perfusion pumps• Multi-flow rate calibration• Flow profile to reduce dead volume delays• Temperature profiling and cycling• Temperature and perfusion recording• Saving and reloading settings• Data logging of perfusion and temperature• Graphic display of events

Graphic represents one of three different control screens:• Profiled Flow (allows control of acceleration rates during
pump speed changes)• Alternating Flow (immediate transition from one pump toanother)• Temperature (programming and recording)This intuitive control interface system is optimized for time-lapse
imaging of live-cell activity. It provides a convenient, accurate,
reliable, and repeatable method of controlling fluid and temperature for chemically or thermally induced change experiments inlive-cell chambers. WYSIWYG on both Mac and PC platforms!
System includes: Program CD, USB Dongle, External USB interface and cabling, Instructions Manual, and Self Help Wall Poster.
Note: Temperature programming and recording of Bioptechs products only. Temperature recording with USB interface only.
System Requirements: USB Mac or USB PC - Windows 2000 orXP, and two Bioptechs Micro-Perfusion pumps (for controlled perfusion) or a Bioptechs Temperature Controller FCS2, FCS3, Delta
T, or Objective Heater (for temperature control).
43

Bioptechs PN 13161603-13 (USB)Perfusion Tubing
It is important to use a tubing suitable for cell culture and compatible with theenvironmental apparatus. The main considerations for tubing are:
•
•
•
•
•

Size
Biocompatibility
Gas permeability
Autoclavability
Transparency

Typically, 1/16th” inside diameter tubing is more than adequate to perfuse the small volumes of cells thatare used on a microscope. The 1/16th” diameter provides adequate flow and is easily workable. There areseveral choices that Bioptechs has found to be suitable for microscopy applications. All have excellent biocompatibility and nearly zero gas permeability. The main differences are the survivability of autoclaving andtransparency. Transparency is an issue when a parallel plate chamber is used. If any bubbles form during
the perfusion process and get into the parallel plate optical cavity, they will adhere to the glass and potentially ruin an experiment. If a transparent tubing is used, the bubbles can be seen, giving you time to trap andeliminate them before they do harm. The final issue is autoclaving. All three of the top biocompatible tubings
claim autoclavability. However, some survive better than others. The following chart represents Bioptechs’
experience in testing these materials.
Material

Size

C-FlexPharmedTygon 2275

1/16”
1/16”
1/16”

Gas Permeability

Transparency

Autoclavability

Translucent
OpaqueNearly transparent

Excellent
Unknown
Fair

Bioptechs PN 20202275 for Tygon (25ft)
Bioptechs PN 2020822 for C-Flex (50ft)

Bioptechs recommends and sells C-Flex tubing. It is available in the 1/16” ID size and sold in 50 foot reels.

• C-Flex is a unique patented thermoplastic elastomer specifically formulated tomeet the critical demands of medical, and pharmaceutical, research.
• C-Flex contains no toxic extractables (non-PVC, non-latex, non-silicone)• Complies with USP XXII, Class VI, FDA and USDA standards.
• Manufactured under strict GMP’s in an FDA registered facility• Significantly less permeable than silicone• Low protein binding• Ultra smooth inner bore
• Safely disposable

44Reference Information
Bioptechs’ Temperature Control System Algorithm
The five input values read by the controller are:
1. User set point
2. Dish temperature below 0.9°C below user setpoint
3. Dish temperature from 0.9°C below user setpoint to 0.1°C below setpoint
4. Dish temperature between user setpoint minus 0.1°C and user setpoint (control resolution)
5. Dish temperature at or above set point
There are six conditions that affect the output:[a] Object temperature is below 0.9°C below user setpoint (cold startup mode). The heater
output starts at 0 volts and increases at a fixed rate to a maximum value.
[b] Object temperature increases above 0.9°C below user setpoint and below setpoint -0.1°C. A
latch is set for condition [f] and output voltage acceleration rate decreases from cold startup
mode and the output voltage is set at a rate about five times the minimum decay rate.
[c] Object temperature is above user setpoint. The output voltage is reduced to one-third its
previous value and decays at an accelerated rate until condition [d] or [e] occurs.
[d] Object temperature falls from above user setpoint to within control resolution band. The
output voltage is held steady at the previous high state value, minus the accelerated decay
value if present, at a minimum decay rate. (Accelerated decay value present only if precededby condition [c].)[e] Object temperature is within control resolution band. The output voltage is held steady but
decays at a minimum rate.
[f] Object temperature or thermistor reading falls below 0.9°C below user setpoint. The output
voltage is shut off to the object and an alarm sounds until the user cancels the alert by pressing the red button. This occurs if the temperature sensor is subject to cold shock. The amplitude, gain and decay rate, maximum current, operating current, response time, sensitivity, alarm range, and bandwidth are all variables thatare adjusted specifically for each type of controller. Although each type of controller auto-ranges, its range ofaccommodation is limited to the typical thermal load of the object it is regulating. Each channel is controlled
separately but both share the same setpoint.
Industry Standard Thickness for Coverslip Glass:#00 = 0.060 - 0.080#2 = 0.170 - 0.250#0 = 0.080 - 0.120#3 = 0.280 - 0.320#1 = 0.130 - 0.170#4 = 0.380 - 0.420#1.5 = 0.160 - 0.190#5 = 0.500 - 0.600

Microscope Stage Types:
Single plate stage is defined by a thick metal plate that uses a separate mechanical specimentranslator to slide the specimen on the surface of the stage. Friction occurs between the specimen and surface of the stage. The specimen sometimes is placed into an adapter plate.
Triple plate stage is made of three individual plates; bearing mounted, to eliminate friction
between the plates. The specimen is placed in or on the top plate and both the specimen andthe top plate move together. This is a premium stage with the best X, Y, Z stability.
Motorized Stages are usually of the triple plate type and provide automation and superior
repeatability.

45

Micro-Environmental System Profile Questions
This list of questions is fundamental to identifying the appropriate components of a micro-environmental system. Completion of this list prior to contacting Bioptechs will optimize the exchange of technical information.
Please copy and fax this report to Bioptechs prior to contacting us by phone. Fax 724-282-0745
Name: ________________________________ Institution: ________________ Phone: ________________
Fax: ______________________ Email: _______________________________
1. Who is the brand & model of microscope? Carl Zeiss Leica Nikon Olympus

Model: _____________

2. What type of microscope?

Upright

Inverted

3. What is the brand of the stage?

Carl Zeiss, Leica, Nikon, Olympus, Marzhouser, Ludl, Prior, ASI, etc.

4. What type of stage for inverted microscopes?

Single plate

5. What mode or combinations of modes of microscopy will be used?
Brightfield
Darkfield
DIC
Polarization
Reflection Interference
FluorescenceStanding Wave
VAREL

Triple plate

Phase
Modulation Contrast
Multi-Photon
Confocal

6. Which types of objectives and their magnifications? (include all that apply)
Inverted microscopes:
4X, 5X, 10X, 20X, 40X dry, (40 X, 63X, and 100X fluid coupled)
Upright microscopes:
4X, 5X, 10X, 20X, (other magnifications require water dipping lenses for Delta T use)
7. What is the condenser N.A.? (relates to working distance) Common values: 0.3-0.6,
8. What is the imaging scenerio?

Single image

0.7-0.9,

1.0 - 1.4

Time lapse (interval) __________________

9. What is the duration of time specimens will be on the microscope? (minutes, hours, days) _________
10. How will the correlation of optical contrast images to other modes such as fluorescence be recorded?
No contrast images
First image only
Contrast image with every fluorescence image
11. What type of experiment or object of experiment? (brief explanation)

12. What is the specimen type?
Adherent monolayer
Cell suspension

Natural tissue

Artificial membrane

13. What temperature do the specimens need to be maintained?
14. What is the appropriate chamber type?

_______°C

Open

Closed

15. Will micromanipulators be used and when? Before imaging

During imaging

Both

16. Does the specimen need perfusion of media and what is the perfusion profile?
Intermittent (manual)
Automated
Continuous
Single or multiple perfusate sources
Explain:
17. Will CO2 dependent media be used or other gas regulation be necessary?
46

CO2 ____, Gas reg. _____

Check List for Ordering an FCS2® or FCS3®
Having established the need for a closed system, there are a few technical matters that should be considered. Following
this guide will ensure that these areas have been addressed.
The FCS2 and FCS3 are used for both high and low numeric aperture applications. Therefore, the objective heater andobjective temperature controller are sold separately. The main optional consideration is perfusion.

When Ordering an FCS2 or FCS3 System:
1. Select a stage adapter from the FCS2 stage adapter page and provide the microscope manufacturer’s
name and model of stage to confirm identification of the appropriate adapter. (See pg 28) If ordering FCS3 skip to #2.
2. If the cells need perfusion, the following items should be included on the order:• Micro-Perfusion Pump
Bioptechs PN 060319131616• C-Flex tubing (50ft)
Bioptechs PN 2020822
or• Tygon tubing (25ft)
Bioptechs PN 20202275• Bioptechs Perfusion Connector Kit,
Bioptechs PN 162003-1
Includes: Male Luer adapter
Two Female Luer adapters,“T” adapter & Three-way Luer valve
3. Although the starter set comes with 50 coverslips, you may want to order additional coverslips.
Bioptechs PN 40-1313-0319 (250pk)
4. If high numeric aperture objectives are to be used, you will need to order the following:
Objective Heater Controller
Bioptechs PN 150803---AND--An Objective Heater* (three sizes available)
14 - 22mm diameter
19 - 27mm diameter
23 - 33mm diameter
30 - 37mm diameter

Bioptechs PN 150819-1913
Bioptechs PN 150819
Bioptechs PN 150815
Bioptechs PN 150812* We cannot guarantee that our stock Objective Heater fits all objectives. Therefore we ask that you specify the make,magnification, N.A. and any other attributes of the objectives you intend to use. This step will assist us in determining thecompatibility of our Objective Heater with your optics
5. Indicate the N.A. of the transmitted condenser if it will be used.
6. Supplemental components for FCS2 operation at temperatures below ambient are:•
FCS2 Cooling Adapter (fits all FCS2’s)
Bioptechs PN 060319 0301•

Objective Cooling Collar(individually machined to fit users own objective)

Bioptechs PN 150303•

Objective Thermal-Isolator(Available for infinity corrected objectives
with an anti-reflective coated window,
160mm tube-length objectives require acompensating lens at an additional charge)

For RMS Thread:
Bioptechs PN 152009R - Isolator
Bioptechs PN 152019R - Spacer
For 25mm x 0.75mm Thread:
Bioptechs PN 152009 - Isolator
Bioptechs PN 152019 - Spacer

47

Check List for Ordering a Delta T® Culture Dish System
Having established the need for an open dish system, there are a few technical matters that should be considered.
Following this guide will ensure that these areas have been addressed.

When Ordering a Delta T System:
1. Provide the microscope manufacturer’s name and model of your stage to identify the appropriate adapter.
2. Indicate the type of specimen.
a. In the case of adherent cultured cells and low N.A. objectives, you will only need the basic Delta
T system. It is comprised of a Delta Stage Adapter and a Delta T Dish Controller plus asuitable quantity of 0.5mm dishes.
b. If the specimen is a natural membrane, cut section, or tissue slice, an appropriate specimen
carrier will need to be added to the basic system. (see Tissue Slice, Brain Slice, and Artificial
Membrane pages).
3. Determine if high or low N.A. lenses will be used. Order the appropriate dishes for the N.A. of theobjectives.
• For low N.A. applications, order 0.5mm thick bottomed dishes, Bioptechs PN 04200405• For high N.A. applications, order #1.5 coverslip bottomed dishes, Bioptechs PN 04200415
Note: If the application involves Fluorescence, add the suffix B to specify black sidewall dishes.
Clear dishes are shipped with clear lids & black dishes are shipped with black lids.
4. If the cells need perfusion, the following items should be included on the order:
If media surface can be open to atmospheric air use:
Delta T Hinged Perfusion Adapters
Bioptechs PN 0420081601
Delta T Micro-Perfusion Pump
Bioptechs PN 0420131616
C-Flex tubing (50ft)
Bioptechs PN 2020822
orTygon tubing (25ft)
Bioptechs PN 20202275
If media must be closed to atmospheric exposure:
Delta T Perfusable Coverglass Lid :Bioptechs PN 0420031216

5. If contrast enhancement techniques are used for transmitted images on an inverted microscope,consider the option of a Heated Lid:
Bioptechs PN 04200803
Perfusable Heated Lid
Bioptechs PN 0420080316
6. If a series of optically induced contrast images are to be made into a movie loop, the Coverglass Lid, withor without perfusion, is advisable.
Delta T Coverglass Lid
Bioptechs PN 04200312
Delta T Perfusable Coverglass Lid
Bioptechs PN 0420031216
7. If high numeric aperture objectives are to be used at physiological temperatures, you will need to order thefollowing:
Objective Heater Controller
Bioptechs PN 150803----AND---An Objective Heater* (four sizes available)
14 - 22mm diameter
19 - 27mm diameter
23 - 33mm diameter
30 - 37mm diameter

Bioptechs PN 150819-1913
Bioptechs PN 150819
Bioptechs PN 150815
Bioptechs PN 150812* We cannot guarantee that our stock Objective Heater fits all objectives. Therefore, we ask that you specify
the make, magnification, N.A. and any other attributes of the objectives you intend to use. This step willassist us in determining the compatibility of our Objective Heater with your optics.
If the temperature of the specimen needs to be below ambient, include the Delta T Cooling Ring attachment.
Bioptechs PN 04200318

48

How to Size Your Objective
A is the diameter of the spring loaded retractable portion of the objective. This is the diameter that the heater
band must fit.
B is the minimum vertical surface needed to mount the Objective Heater and transfer heat.

Bioptechs Objective Heater Band Sizes:
SmallStandard

MediumLarge

A - 14 - 22 mm

A: 19 - 27 mm

A: 23 - 33 mm

A: 30 - 37 mm

B - 5.5 mm

B: 5.5 mm

B: 6mm

B: 3.5 mm

Bioptechs PN 150819-1913

Bioptechs PN 150819

Bioptechs PN 150815

Bioptechs PN 150812

Please note that the above diagram depicts a typical Objective Heater mounting location. There are a wide
variety of objective configurations. Therefore it may be necessary to mount an Objective Heater at a different
location than indicated above. If the geometry of your objective will not accommodate an Objective Heater,
contact us via email (info@bioptechs.com) or fax us with the information below. Please add your contact information and we will give you a definite answer as to whether your objective can accept an Objective Heater.
Please send faxes, Attn: Sales, Fax: (724)282-0745.
Your measurement for Dimension A
Your measurement for Dimension B
Your Objective Manufacturer:Your Objective Specifications:

mm or
mm or

inches.
inches.

Some objectives require the removal of a decorative collar as shown below. If removal of a decorative collar isrequired an objective wrench and pliers are available from Bioptechs.

Objective Wrench and Pliers
Bioptechs PN 021523
Objective

Inside of
Objective

Outer Shieldof Objective

Objective with
Heater Installed49

Bioptechs Delta T® Price Schedule
Product

Part Number

Domestic

ExportDelta T Starter Set Includes the following:

0420-4$3,350.00$3,852.50

Delta T Controller, Heated Lid, Coverglass Lid, Culture Cylinder Pack: (1) 4mm, (1) 6mm, (1) 8mm, (1) 10mm, 0.17mm
clear dishes (10/pk), Standard Stage Adapter (please specify which stage when ordering)

Delta T Culture Dish System (Page 14)
Delta T Culture Dish Controller
0420-4-03
Delta T Culture Dish Stage Adapters (Page 25)
96-Well Plate Sized Adapter
04202003
Leica DMIRB Triple Plate Adapter
04201203
Nikon, Olympus, Deltavision Adapter
04201415
Upright Universal Adapter
04202116Marzhauser, Zeiss (K), ASI, Ludl & Prior Adapter
04202602
Zeiss (G) Adapter
04202603
Zeiss (M) & Leica Single Plate Adapter
04202601
Custom Delta T Stage Adapters:
Cell Robotics Adapter
04201220
Leica Galvanic Adapter (Round)
04201207
Leica Galvanic Adapter (Rectangular)
04201807
Marzhauser Galvanic Adapter (Rectangular)
04202807
Delta T Culture Dishes (10/pk) (Page 15)
0.5mm thick glass (clear)
04200405
0.17mm thick glass (clear)
04200415C
0.17mm thick glass (black)04200415B
Delta TPG Uncoated (no temperature control) Culture Dishes (10/pk)
0.5mm thick glass
0420040500
0.17mm thick glass
0420041500C
0.17mm thick glass
0420041500B
Delta T Culture Dish Accessories (Page 17)
Tissue Slice Adapter
0420201918
Artificial Membrane Adapter
0420011301
Brain Slice Adapter
0420201919
Hinged Perfusion Adapter Set (2/set)
0420081601
Cooling Ring
04200318
Heated Lid
04200803
Perfusable Heated Lid
0420080316
Atmospheric Barrier Ring
04200325
Coverglass Lid
04200312
Perfusable Coverglass Lid
0420031216
Media Volume Reducer
04200303
Yeast Slide Adapter
042005076
Perfusion (Page 35)
Delta T Micro-Perfusion Pump
0420131616
Delta T Dual Channel Tubing Assembly (4/set) C-Flex
0420042016
Delta T Dual Channel Tubing Assembly (4/set) Silicon
0420042016S
1/16" Tygon 2275 Tubing (25 ft)
20202275
C-Flex Tubing (50 ft)
2020822
Perfusion Pump Rod Mounting Clamp
16181303
Perfusion/ Temperature Controller (USB interface)13161603-13
Objective Heater System
See Catalog Page 39
Glass Culture Cylinders
See Catalog Page 26
Note: 1. Bioptechs reserves the right to change prices.
2. The prices listed above are stated in US $.
50$2,500.00$2,875.00$550.00$550.00$550.00$550.00$550.00$550.00$550.00$632.50$632.50$632.50$632.50$632.50$632.50$632.50$750.00$750.00$750.00$750.00$862.50$862.50$862.50$862.50$57.50$57.50$57.50$66.13$66.13$66.13$35.00$35.00$35.00$40.25$40.25$40.25$350.00$400.00$400.00$300.00$150.00$300.00$375.00$50.00$150.00$250.00$300.00$500.00$402.50$460.00$460.00$345.00$172.50$345.00$431.25$57.50$172.50$287.50$345.00$575.00$995.00$125.00$125.00$55.00$90.00$25.00$3.200.00$1,144.25$143.75$143.75$63.25$103.50$28.75$3,680.00

Bioptechs FCS2® & FCS3® Price Schedule
Product

Part Number

Domestic

ExportFCS2 System (Page 28)
FCS2 Starter Set (includes:)060319-2$4,000.00$4,600.00

FCS2 Chamber, Chamber Controller, 5 Microaqueduct Slides, 50-40mm Coverslips, and Gasket Set (30/set)

FCS2 Stage Adapters (Page 34)
Cell Robotics Adapter
Leica DMIRB Triple Plate Adapter
Nikon, Olympus, Deltavision Adapter
Zeiss (K), ASI, Ludl & Prior Adapter
Zeiss (M) & Leica Single Plate Adapter
Closed Chamber Supplies
FCS2 Chamber
FCS2 Chamber Controller
Microaqueduct Slides (5/pk)
40mm Coverslips (250/pk)
40mm Coverslips (500/pk)
40mm Coverslips (1000/pk)
FCS2 Gasket Set (30/set)
FCS2 Accessories (Page 32)
FCS2 Cooling Adapter
FCS2 Open Mode Adapter
FCS2 Low Dead Volume Top
Cooled FCS2 (CFCS2)
Perfusion (Page 30)
FCS Micro-Perfusion Pump
FCS Single Channel Tubing Assembly (4/pk) C-Flex
FCS Single Channel Tubing Assembly (4/pk) Silicon
1/16" Tygon 2275 Tubing (25 Ft)
1/16" C-Flex Tubing (50 Ft)
1/16" Perfusion Adapter Set
Micro-Perfusion Pump Rod Mounting Clamp
Perfusion/Temp Control (USB Interface)
FCS3 System (Page 35)
FCS3 Starter Set (includes:)060319-2-0318
060319-2-12060319-2-1415
060319-2-2611
060319-2-2613$330.00$550.00$375.00$650.00$500.00$379.50$632.50$431.25$747.50$575.00

060319-2-03
060319-2-0303
130119-5
40-1313-0319
40-1313-03192
40-1313-03193
060319-2-0719$1,650.00$2,000.00$350.00$200.00$350.00$650.00$150.00$1,897.50$2,300.00$402.50$230.00$402.50$747.50$172.50060319-2-0301
060319-2-1513
060319-2-1242
03060319-2$400.00$175.00$495.00$2,500.00$460.00$201.25$569.25$2,875.00

060319131616
060319192016
060319192016S
20202275
2020822
162003-1
16181303
13161603-13$995.00$125.00$125.00$55.00$90.00$15.00$25.00$3,200.00$1,144.25$143.75$143.75$63.25$103.50$17.25$28.75$3,680.00

21-060319-3$4,000.00$4,600.00

FCS3 Chamber, Chamber Controller, 5 Microaqueduct Slides, 50- #1.5,40mm Coverslips, and Gasket Set (30/set)**Note: FCS2 and FCS3 gaskets, coverslips, and microaqueduct slides are interchangable**

FCS3 Universal Upright Stage Adapter
FCS3 Zeiss A Stage Adapter
FCS3 K Stage Adapter
FCS3 Chamber
FCS3 Chamber Controller
FCS3 Spanner Wrench
FCS3 Pressure Plate
Thermal Slide System
Thermal Slide Starter Set (Includes:)21-060319-3-08
21-060319-3-2601
21-060319-3-2611
21-060319-3-03
21-060319-3-0303
21-060319-3-1923
21-060319-3-16$1,000.00$1,000.00$1,250.00$2,602.00$2,000.00$54.00$36.00$1,150.00$1,150.00$1,437.50$2,992.30$2,300.00$62.10$41.402019-1919-0$1,700.00$1,955.00

Thermal Slide Controller, Controller Stand, Controller Power Supply, Thermal Slide Carrier, (1pk) 5qty Thermal Slides

Thermal Slide Controller w/ Power Supply
Thermal Slide Carrier
Thermal Slide Pack (5 qty)
Note:2019-1919-3
2019-1919-1
2019-1919-2

1. Bioptechs reserves the right to change prices.
2. The prices listed above are stated in US $.

51$1,285.00$375.00$150.00$1,477.75$431.25$172.50

ProductObjective Heater System (Page 39)
Objective Heater Controller
Objective Heater Small
Objective Heater Standard
Objective Heater Medium
Objective Heater Large
37 Degree Immersion Oil
Objective Wrench & Pliers
Objective Thermal Isolator RMS
Objective Thermal Isolator 25mm
Objective Thermal Spacer RMS
Objective Thermal Spacer 25mm
Objective Cooling Collar
Boekel Warmer
Stable "Z" System
Stable Z Starter Set

Part Number

Domestic

Export150803150819-1913
150819150815150812150825021523152009R
152009152019R
152019150303260700$1,500.00$625.00$625.00$665.00$700.00$6.25$175.00$500.00$500.00$100.00$100.00$700.00$250.00$1,725.00$718.75$718.75$764.75$805.00$7.19$201.25$575.00$575.00$115.00$115.00$805.00$316.25403-1926$1,775.00$2,041.25

Includes: Stable Z Controller, Stable Z Controller Holder, Stable Z Controller Power Supply, andStable Z Stage Adapter (Please specify which stage adapter when ordering)

Stable Z Controller
K Stage Adapter
Nikon Stage Adapter
Olympus Stage Adapter
SBS Stage Adapter
A Stage Adapter
Nunc Lab-TekTM Warming Adapter
Replacement Reference Thermistor
Interchangeable Coverslip Dish Set

403-1926-03$1,285.00
403-1926-19-11-K$590.00
403-1926-19-14-N$590.00
403-1926-19-15-O$590.00
403-1926-19-19-S$590.00
403-1926-19-2601-A $590.00
403-1323$825.00
403-1926-1820$21.00190310$80.00$1,477.75$678.50$678.50$678.50$678.50$678.50$948.75$24.15$92.00Includes: Dish Base, 10mm Threaded Insert, (2) o-rings, (50pk) 30mm Coverslips

Interchangeable Coverslip Dish Set (10+)

190310$65.00 per$74.75 perSpecial Pricing When Ordering 10+ Includes: (10+) Dish Base, (10+) 10mm Threaded Insert, (20+) O-rings,(500+pk) 30mm coverslips

30mm Coverslips (#1.5) Qty 100
EDU Culture Dish Control System
EDU Starter Set

301313-0319$27.00$31.050420-050421-2$1,175.00$1,351.25

Includes: EDU Controller, EDU Controller Power Supply, EDU Stage Adapter, (1pk) 10qty clear dishes, (1pk)
100 qty 22mm coverslips, 22mm Glass Spacer, Coverslip Weight, 6mm Culture Cyl

EDU Controller w/ Power Supply
EDU Stage Adapter
Glass Culture Cylinders (Page 26)
Starter Set (2, 4, 6, 8, 10 mm)
1 mm Inner Diameter x 5mm high
2 mm Inner Diameter x 5mm high
4 mm Inner Diameter x 5mm high
6 mm Inner Diameter x 5mm high
8 mm Inner Diameter x 5mm high
10 mm Inner Diameter x 5mm high
12 mm Inner Diameter x 5mm high
14 mm Inner Diameter x 5mm high0420-050421
0420-050421-1$600.00$400.00$690.00$460.00

070303-1919
070303-01
070303-02
070303-04
070303-06
070303-08070303-10070303-12070303-14$125.00$25.00$25.00$25.00$25.00$25.00$25.00$25.00$25.00$143.75$28.75$28.75$28.75$28.75$28.75$28.75$28.75$28.7552

References
This is not a complete list of Bioptechs' references, a keyword searchable database of these and many other references areavailable on the Bioptechs web site. www.bioptechs.com
Title

Citation

Authors

The Zn2+ transporting pathways in pancreatic beta-cells: A
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J. Biol. Chem. published 30 December 2005, 10.1074/
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Armen V. Gyulkhandanyan, Simon C. Lee, George Bikopoulos, Feihan Dai, and Michael B. Wheeler

Distinct Morphological Stages of Dentate Granule Neuron
Maturation in the Adult Mouse Hippocampus

J. Neurosci. 2006; 26(1): p. 3-11

Chunmei Zhao, E. Matthew Teng, Robert G. Summers, Jr,
Guo-li Ming, and Fred H. Gage

A transport mechanism for NAADP in a rat basophilic cellline

FASEB J. published 10 January 2006, 10.1096/fj.05-5058fje

R. A. Billington, E. A. Bellomo, E. M. Floriddia, J. Erriquez,
C. Distasi, and A. A. Genazzani

A role for the septation initiation network in septum assembly revealed by genetic analysis of sid2-250 suppressors

Genetics published 16 January 2006, 10.1534/
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Quan-wen Jin, Mian Zhou, Andrea Bimbo, Mohan K Balasubramanian, and Dannel McCollum

Kinetics of penetration influence the apparent potency ofvanilloids on TRPV1

Mol. Pharmacol. published 17 January 2006, 10.1124/
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Jozsef Lazar, Derek C. Braun, Attila Toth, Yun Wang, Larry
V. Pearce, Vladimir A. Pavlyukovets, Peter M. Blumberg,
Susan H. Garfield, Stephen Wincovitch, Hyun-Kyung Choi,
and Jeewoo Lee

Regulation of N-Cadherin Dynamics at Neuronal Contacts
by Ligand Binding and Cytoskeletal Coupling

Mol. Biol. Cell. 2006; 17(2): p. 862-875

Olivier Thoumine, Mireille Lambert, Rene-Marc Mege, andDaniel Choquet

Cell Cycle-dependent Recruitment of Telomerase RNA and
Cajal Bodies to Human Telomeres

Mol. Biol. Cell. 2006; 17(2): p. 944-954

Beata E. Jady, Patricia Richard, Edouard Bertrand, andTamas Kiss

Localized recruitment and activation of RhoA underlies
dendritic spine morphology in a glutamate receptor-dependent manner

J. Cell Biol. 2006; 172(3): p. 453-467

Vanessa Schubert, Jorge Santos Da Silva, and Carlos G.
Dotti

A functional module of yeast Mediator that governs the
dynamic range of heat shock gene expression

Genetics published 1 February 2006, 10.1534/
genetics.105.052738

Harpreet Singh, Alexander M Erkine, Selena B. Kremer,
Harry M Duttweiler, Donnie A Davis, Jabed Iqbal, Rachel R
Gross, and David Gross

Disruption of SLP-76 Interaction with Gads Inhibits Dynamic
Clustering of SLP-76 and Fc{varepsilon}RI Signaling in
Mast Cells

Mol. Cell. Biol. 2006; 26(5): p. 1826-1838

Michael A. Silverman, Jonathan Shoag, Jennifer Wu, andGary A. Koretzky

CLIP-170 Homologue and NUDE Play Overlapping Roles in
NUDF Localization in Aspergillus nidulans

Mol. Biol. Cell published 8 February 2006, 10.1091/
mbc.E05-11-1084

Vladimir P. Efimov, Jun Zhang, and Xin Xiang

The beta-agonist isoproterenol attenuates EGF-stimulated
wound closure in human airway epithelial cells

AJP: Lung. 2006; 290(3): p. L485-L491

Bradley J. Schnackenberg, Stacie M. Jones, Crystal Pate,
Brian Shank, Laura Sessions, Laura M. Pittman, Lawrence
E. Cornett, and Richard C. Kurten

Myo1c binds tightly and specifically to phosphatidylinositol
4,5-bisphosphate and inositol 1,4,5-trisphosphate

Proc. Natl. Acad. Sci. USA published 21 February 2006,
10.1073/pnas.0505685103

David E. Hokanson and E. Michael Ostap

PKG phosphorylates Pak1, inhibiting Pak/Nck binding andstimulating Pak/VASP association

J. Biol. Chem. published 20 February 2006, 10.1074/
jbc.M600279200

Benjamin H. Fryer, Changhui Wang, Srilakshmi Vedantam,
Guo-Lei Zhou, Shenghao Jin, Linda Fletcher, M. Celeste
Simon, and Jeffrey Field

Comparative Evaluation of HERG Currents and QT Intervals
following Challenge with Suspected Torsadogenic andNontorsadogenic Drugs

J. Pharmacol. Exp. Ther. 2006; 316(3): p. 1098-1106

Alexander N. Katchman, John Koerner, Toshimasa Tosaka,
Raymond L. Woosley, and Steven N. Ebert

Rap1-Mediated Activation of Extracellular Signal-Regulated
Kinases by Cyclic AMP Is Dependent on the Mode of Rap1
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Zhiping Wang, Tara J. Dillon, Viji Pokala, Snigdha Mishra,
Kirstin Labudda, Brian Hunter, and Philip J. S. Stork

Changes in chromatin structure and mobility in living cells atsites of DNA double-strand breaks

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Michael J. Kruhlak, Arkady Celeste, Graham Dellaire, Oscar
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Genetic ablation of zyxin causes Mena/VASP mislocalization, increased motility, and deficits in actin remodeling

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Laura M. Hoffman, Christopher C. Jensen, Susanne
Kloeker, C.- L. Albert Wang, Masaaki Yoshigi, and Mary C.
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Assembling an intermediate filament network by dynamic
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Lynne Chang, Yaron Shav-Tal, Tatjana Trcek, Robert H.
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A motor neuron disease-associated mutation in p150Glued
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Jennifer R. Levy, Charlotte J. Sumner, Juliane P. Caviston,Mariko K. Tokito, Srikanth Ranganathan, Lee A. Ligon,
Karen E. Wallace, Bernadette H. LaMonte, George G.
Harmison, Imke Puls, Kenneth H. Fischbeck, and Erika L.F.
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Synthetic D-amino acid peptide inhibits tumor cell motility
on laminin-5

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Role of target geometry in phagocytosis

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Julie A. Champion and Samir Mitragotri

A physical model of axonal damage due to oxidative stressProc. Natl. Acad. Sci. USA published 28 March 2006,
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Anne E. Counterman, Terrence G. D’Onofrio, Anne Milasincic Andrews, and Paul S. Weiss

Cholesterol Is Required for Efficient Endoplasmic Reticulum-to-Golgi Transport of Secretory Membrane ProteinsMol. Biol. Cell. 2006; 17(4): p. 1593-1605

Andrew Ridsdale, Maxime Denis, Pierre-Yves Gougeon,
Johnny K. Ngsee, John F. Presley, and Xiaohui Zha

Auto-activation of the apoptosis protein BAX increases mitochondrial membrane permeability and is inhibited by BCL-2

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Chibing Tan, Paulina J. Dlugosz, Jun Peng, Zhi Zhang,Suzanne M. Lapolla, Scott M. Plafker, David W. Andrews,
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The Zn2+-transporting Pathways in Pancreatic beta-Cells:
A ROLE FOR THE L-TYPE VOLTAGE-GATED Ca2+CHANNEL

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A Coiled-Coil Domain of Melanophilin Is Essential for
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Alistair N. Hume, Abul K. Tarafder, Jose S. Ramalho, Elena
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This is not a complete list of Bioptechs' references, a keyword searchable database with these and hundreds of otherreferences are available on the Bioptechs web site. www.bioptechs.com

54

Notes55