Count Cells Without Slides
CellDrop FL, CellDrop BF, CellDrop FLi,
CellDrop FLxi, CellDrop BFx

Version: 1-AUG-20241

1.

Introduction

4

Product Technology ........................................................................................................................ 4
Key Features ...................................................................................................................................4
Optics .............................................................................................................................................. 4

2.

Set-up and Safety

63.

Instrument Diagram 8

4.

Quick Guide

9

Cell Count Procedure ...................................................................................................................... 9Best Practices ............................................................................................................................... 10

5.

General Operation and Software Information

11

Top Action Bar .............................................................................................................................. 11
Software Icons and Buttons .......................................................................................................... 12
Chamber Height Options............................................................................................................... 12
Data Storage and Memory Warnings ............................................................................................ 13
Channels ....................................................................................................................................... 14
Count Apps ...................................................................................................................................15
Brightfield ...................................................................................................................................... 15
Trypan Blue Tissue Culture........................................................................................................... 15
Trypan Blue Primary Cells ............................................................................................................ 15
Trypan Blue Nuclei ........................................................................................................................ 15
Erythrosin B Tissue Culture .......................................................................................................... 15
Erythrosin B Primary Cells ............................................................................................................ 15
Erythrosin B Nuclei........................................................................................................................ 16
AO/PI............................................................................................................................................. 16
AO ................................................................................................................................................. 16
PI ................................................................................................................................................... 16
Yeast ............................................................................................................................................. 16
Yeast AO/PI ..................................................................................................................................17
Yeast Brightfield ............................................................................................................................ 17
GFP ............................................................................................................................................... 17
Hepatocytes ..................................................................................................................................17
Primary Cell AO/PI ........................................................................................................................ 18
Nuclei AO/PI..................................................................................................................................18
Fixed Cells PI ................................................................................................................................ 18
Fixed Nuclei PI .............................................................................................................................. 18
Custom Methods ........................................................................................................................... 186.

Count Tab View

19

Sample Name and Sample Information ........................................................................................ 19
Image Controls .............................................................................................................................. 19
Dilution Calculator ......................................................................................................................... 21
Count Button .................................................................................................................................21

7.

Count Settings 22

8.

Results Tab View

25

Export Icon ....................................................................................................................................25
Next Count ....................................................................................................................................25
Common controls .......................................................................................................................... 25
Export Icon ....................................................................................................................................26
Discard. New Count ...................................................................................................................... 26
Save and e-Sign............................................................................................................................ 26
Export and Backup of Signed Record ........................................................................................... 27
Controls After e-Signing Record ...................................................................................................27
Signed. Next Count ....................................................................................................................... 28
Add Signature ............................................................................................................................... 28
Record Security............................................................................................................................. 28

9.

Result Tab Common Controls

29

Optimize Settings .......................................................................................................................... 29
Cell Size Gating ............................................................................................................................ 292

Intensity Gating (GFP app) ........................................................................................................... 29
Action Bar Controls ....................................................................................................................... 30
Results View Options .................................................................................................................... 30

10. Export and Print Options

32

Screen Capture ............................................................................................................................. 32
Export ............................................................................................................................................ 32

11. Utility Apps

33

User Accounts Standard Mode ..................................................................................................... 33
User Accounts 21 CFR Mode ....................................................................................................... 34
Data............................................................................................................................................... 36
Count Codes .................................................................................................................................37
Calculator ...................................................................................................................................... 38
Timer ............................................................................................................................................. 38
Settings ......................................................................................................................................... 38
Updater ......................................................................................................................................... 41
ScreenLocker ................................................................................................................................ 41

12. 21 CFR App

42

21 CFR .......................................................................................................................................... 42
Archiving ....................................................................................................................................... 46

13. Troubleshooting and Maintenance48

Quick Help..................................................................................................................................... 48
Software Errors ............................................................................................................................. 48
Cleaning ........................................................................................................................................ 48
Solvent Compatibility..................................................................................................................... 49

14. Customer Support

50

Warranty........................................................................................................................................ 50
Instructions for Disposal of Packaging and Waste Equipment...................................................... 50
Research Use Only ....................................................................................................................... 50

3

1. Introduction
Product Technology
DeNovix CellDrop™ Automated Cell Counters are compact instruments with integrated processors andhigh-resolution touch screens. The software utilizes application specific icons along with a custom
operating system to provide an intuitive, easy-to-use instrument for cell count measurements.
The pre-configured EasyApps™ software is pre-installed and factory optimized for ease of use. The
software allows the user to customize settings when needed to meet lab specific workflow preferences.
The CellDrop uses three chamber heights to enable cell counts over a wide dynamic concentration range
according to specifications. This innovative, patented technology (see denovix.com/patents) along withuser-defined protocols allows users the flexibility required for today’s cell counting workflows and allows
users to Count Cells Without Slides.
This User Guide is designed for both CellDrop Brightfield and CellDrop Dual Fluorescence plus Brightfieldmodels. Several applications and software features are only applicable for instruments with Fluorescencemode and do not apply to the CellDrop Brightfield models. In addition, some applications are only
applicable for CellDrop FLxi or CellDrop BFx which include higher power magnification.

Key Features•

No consumable slides required•Stand-alone (no PC required) with integrated processor and Hi-Definition 7-inch touchscreen•

Live preview of sample and immediate, on-screen analysis of count result•

Custom software interface with intuitive EasyApps software•

Variable chamber height to enable an accurate count for a broad range of cell population
densities

OpticsBrightfield•

Illumination = LED 530 nm

Fluorescence•

Excitation: 470 nm +/- 20 nm•

Green Fluorescence Emission: 525 nm +/- 25 nm•

Red Fluorescence Emission 645 nm +/- 37 nm

Upgrade Options
CellDrop instruments may be upgraded at any time. Contact DeNovix or your distributor if you areinterested in the following:•

Convert Pay-As-You-Go to Unlimited•

Convert CellDrop Brightfield models to CellDrop Dual Fluorescence plus Brightfield models•

Activate 21 CFR Part 11 Compliance Ready Software•

Activate machine learning apps

4Models and Applications
CellDrop is available in multiple models which should be selected based on the main application forwhich the instrument will be used. Note that CellDrop FLi replaced CellDrop FL as the DeNovix
standard Dual Fluorescence plus Brightfield instrument.

5

2. Set-up and Safety
1.

Remove all packing materials including materials protecting the arm and sample surfaces.

2.

Place the instrument on a flat bench and avoid vibrations to ensure optimal performance.

3.

Plug the unit into a 100-240 VAC, 50-60/Hz receptacle using the grounded power supply (GlobTek,
Inc. part number TR9CE3000C95CP-NR6B, 12V / 3A / 36W) included with each unit.

4.

Toggle the power switch to the On position to turn on the instrument. To complete initialization,
ensure that the arm is down and no materials are between the sample surfaces. The operating
system will boot up in less than a minute. Toggle the power switch to Off to initiate shutdown of the
instrument.

Biological Hazard Safety
Trypan Blue, Acridine Orange, Propidium Iodide and other reagents are known hazardous materials.
Consult Safety Datasheets (SDS) for these assays prior to use.
Biological samples such as tissues and bodily fluids of humans and other animals have the potential totransmit infectious diseases. Follow all applicable local regulations. Wear appropriate personal protective
equipment (PPE). Read and follow the guidelines in this User Guide, in the SDS for any chemicals in use
and additional biosafety publications applicable for the types of sample used and institution/region of use.

Instrument Connectivity
The CellDrop comes equipped with Wi-Fi, Ethernet, HDMI and three USB ports. The USB ports are used
to facilitate data transfer to a FAT32 formatted USB storage device and support the use of an Opticon OPI
3601 USB barcode reader. USB mouse and keyboard devices may also be used with CellDrop Series
instruments. The CellDrop does not support printers connected directly to the device via USB.
Cautions
DO NOT REMOVE COVER: No operator serviceable components inside. Refer servicing to qualified
personnel. CellDrop Cell Counters are designed for indoor use under the following conditions:Temperature: 15˚ to 35˚ C | Humidity: 35 to 65 % | Elevation: 2000m or less | Pollution: Pollution degree2
Use only the power supply provided with the instrument. Use of the instrument in any manner not specified
by the manufacturer may impair the protection provided by the supplied power cord and power supply.
NE PAS OUVRIR L’APPAREIL.
Aucun composant réparable par un utilisateur est inclus. Confiez l'entretien à du personnel qualifié. Le
CellDrop est conçu pour une utilisation en intérieur dans les conditions suivantes :Température : 15° à 35˚C | Humidité: ........35 à 65% | Élévation: 2000m ou moins| Pollution: Degré de pollution 2
Utilisez uniquement le bloc d'alimentation inclus avec l'instrument. Utiliser l’appareil dans des conditions
non spécifiées par le fabricant peut compromettre la protection offerte par le cable d’alimentation et le bloc
d'alimentation fournis avec l’appareil.

6

VCCI (Class B) compliance statement for users in Japan:

This is a Class B product based on the standard of the Voluntary Control Council for Interference (VCCI)
from Information Technology Equipment. If this is used near a radio or television receiver in a domestic
environment, it may cause radio interference. Install and use the equipment according to the instruction
manual. This equipment has been tested for radio frequency emissions and has been verified to meet
CISPR 22.2 Class B.

7

3. Instrument Diagram8

4. Quick Guide
Cell Count Procedure1.

Launch any EasyApps Count app.

2.

Lower arm to start live preview. Check to ensure that the field of view is clear.

3.

Select a protocol.
Optional: Enter sample name and additional sample info.
Optional: Edit the Protocol Count Settings for the sample being measured. Check that the protocol
includes the correct Dilution Factor.

4.

Mix the sample well. Load the sample volume indicated on the Count button. Slide the pipette tip
along the alignment groove until it rests against the top surface. Dispense the sample into thecounting chamber formed between the upper and lower sample surfaces.

5.

Allow the cells to settle.

6.

Touch the focus icon and then adjust while viewing the Brightfield channel. Live cells should have abright center and a dark outline. The image below on the left is correctly focused. The image on theright is out of focus. Verify focus in green channel for AO, AO/PI and Yeast apps.

7.

Adjust Trypan Exposure: Touch the exposure icon and select Normal or Low exposure. Live cells
should have a white center with a dark ring like the image below on the left. The middle image isoverexposed and the right image is underexposed.

8.

Fluorescence Exposure: Select the green or red fluorescence channel. Touch the exposure icon
and then adjust the setting. The fluorescence signal should not extend outside of the boundaries of
the cell when viewed in brightfield. The image on the left is correct. The middle image isoverexposed and the right image is underexposed. Adjust the exposure for each channel
independently.
99.

Press Count.

10. Review the result.
Optional: Adjust gating to exclude debris or alternative cell populations from analysis.
Optional: Press Optimize Settings to adjust count settings and reanalyze the image(s).
11. Once a measurement is complete, lift arm and wipe away sample with a clean, dry laboratory wipeand close the arm. When the arm is lifted, the result is immediately saved. Pressing Count again
prior to lifting the arm will overwrite the previous result for that sample.
12. Results from the active measurement session may be viewed while the app is open or viewed later
using the Data app. Saved results may be reanalyzed if desired.
13. Export results as desired.

Best Practices
Note: See Technical Note 186 – CellDrop Best Practices for additional details.
•

Ensure that surfaces are clean prior to loading a sample into the sample chamber.

•

If a previous sample has dried on the surfaces or if the sample is not loading easily, clean thesample surfaces. Lower arm and pipette 15 µL of 70% ethanol into the alignment groove andsample chamber. Wait 10 seconds then firmly wipe both upper and lower surfaces with a drylaboratory wipe.

•

Vortex samples before loading.

•

Ensure that all sample concentrations fall within concentration limits of the instrument for accurate
results.

•

Use the appropriate protocol settings for Chamber Height when measuring high or low
concentration samples or when using slides.

•

Avoid introducing air bubbles into the sample solution when mixing or dispensing samples.

•

Avoid drawing sample back into pipette tip after dispensing.

•

Refer to DeNovix Assay protocols or other manufacturer guidelines for assay specific instructions.

10

5. General Operation and Software Information
CellDrop Automated Cell Counters include pre-installed EasyApps software for rapid counting andviability assessment. Each app is designed for a common cell counting technique such as Brightfield,
Trypan Blue, AO/PI, etc.
Activation of 21 CFR Part 11 Compliance Ready Software affects several aspects of the userinterface. Where applicable in this guide, additional instructions for 21 CFR Mode operation areprovided. A license and activation code must be purchased to activate these features. Contact
DeNovix or your local Distributor. 21 CFR Mode requires users to login to the device.
Apps include a common Top Action Bar that allows a user to easily navigate between the live
preview on the Count tab and the Result tab. Controls available from the action bar are described
below.

Top Action Bar•

Count Tab: Select to view a live preview on screen. View the loaded sample, select saved
protocols, edit protocol settings, define dilution guidance parameters and initiate a cell count.

•

Results Tab: This tab enables the user to review the data for all samples measured in thecurrent measurement session.

•

Alert: The alert icon (bell) is displayed to notify the user of system alerts such as timer
expiration or low count alerts (Pay-As-You-Go models).

•

Account: The current account in use is displayed on the top action bar. The General accountis the default account for CellDrop applications. Tap the account name to display a list ofavailable accounts. Changing an account will automatically use the protocol indicated as thedefault option selected in the User Accounts app.
o 21 CFR Mode: The account that is actively logged in is displayed. The default accounts“Primary Admin” and “General” are disabled and may not be used.

•Home Icon: Tapping the Home icon will navigate back to the Home screen and close the app
and current measurement session. All previous results are saved and accessible from theData app.

•

Overflow: The Overflow icon is used to access a menu of screen specific options:o Screen Capture: Screen shots may be taken of most but not all views. Refer to the Export
and Print Options chapter for more information.
o Count Code Log (PAYG models only): Displays a log of count code use.
o Tech Support: Sends an email with diagnostic information to DeNovix. Email must be
preconfigured from the Settings app for this function to work correctly.
o Set Focus: Allows the user to save a global focal position for DirectPipette™ mode or for
slide mode. This position is the initial position used when opening a count application andessentially eliminates the need to adjust focus once set for CellDrop.
o Reset Auto Focus: Used to clear the saved auto focus starting position allowing the user
to rerun the algorithm to define a best starting position.
o About: The About feature brings up an information box displaying the current app version
number.

11

Software Icons and Buttons
CellDrop applications include a set of standard icons and buttons used throughout the software. The
following table defines each button.
Home

Add

Select All

Overflow
Menu

Edit

Increase /Decrease

Count

Copy

ExportExposure

DeleteClear
Selected

Focus

Expand /Collapse

Information

Calculator

Notifications

Favorite

Chamber Height Options
CellDrop Cell Counters use three chamber heights to enable measurements over a wide cell density
range. The 100 um chamber height is equivalent to the 0.1 mm depth used on standard
hemocytometers.
The default chamber height is 100 um and is suitable for most counts. Users may choose a different
chamber height based on the anticipated density. The upper surface position is automatically moved
to the position defined in the protocol. The tables below are a guideline of the best chamber height tochoose for each CellDrop model based on the density of the sample.

Standard Magnification (FLi & BF)
Gap Height(um)Volume(uL)

Minimum Density(cells/mL)

Maximum Density(cells/mL)

400

40

7.0E+02

3.1E+06100

10

2.9E+03

1.3E+0750

55.9E+03

2.5E+07

Gap Height(um)Volume(uL)

Minimum Density(cells/mL)

Maximum Density(cells/mL)

400

40

4.3E+03

2.6E+07100

10

1.7E+04

1.0E+0850

53.4E+04

2.1E+08

Higher Magnification (FLxi & BFx)

12

Data Storage and Memory Warnings
CellDrop models include a solid-state drive with 120 GB or more of storage capacity, sufficient forseveral thousand results. The user will be alerted if the memory approaches capacity. Data can bedeleted from the CellDrop at any time from the Data App (Standard Mode) or Archived (21 CFRmode). If the Memory Full warning is activated, CellDrop will not save any more results. The user or
System Administrator must make sufficient memory available to make additional counts. Memory
upgrade options are available. Contact DeNovix or your DeNovix Distributor for details.

Display Sleep
The backlight for the screen will automatically turn off after a user-defined period of inactivity. Touchthe screen to turn it back on. Adjust the Sleep Time from Display section in the Settings App.

Preview Suspend
The CellDrop live preview on the Count screen is suspended if the arm is left in the down position for
longer than two minutes. The user may resume the preview when desired.

13

Count Apps
Count apps are designed for a common cell counting techniques such as Brightfield, Trypan Blue, AO/PI,
etc. To properly count a sample and optimize a result, the researcher must first capture the best image(s)possible. Examples of proper and improper adjustments are included in this guide in Section 6, Count Tab.
Protocols and Optimizing Settings
Each Count app includes the relevant group of count settings saved as a Protocol. Once a sample has
been counted, protocol parameters can be adjusted using the Optimize Settings button.

Channels
Count apps use an application specific set of optical channels. The CellDrop FL models include Brightfield,
Green Fluorescence and Red Fluorescence channels. The CellDrop BF models include Brightfield only.

Apps

ModelsBF

FL

FLi

Channels
FLxi

BFx

Brightfield
Trypan Blue
Tissue Culture
Trypan Blue
Primary Cell
Trypan Blue
Nuclei
Erythrosin B
Tissue Culture
Erythrosin B
Primary Cell
Erythrosin B
NucleiBrightfield

Green

Red

-

-

-

-

-

-

-

-

-

-

-

-

-

-

AO/PI

-

-

AO

-

-

PI

-

-

Yeast

-

Yeast AO/PI

-

-

-

Yeast
Brightfield

-

-

-

GFP

-

Hepatocytes-

AO/PI Primary--

AO/PI Nuclei--

Fixed Nuclei

-

-

-

Fixed Cells

-

-

-

CustomMethods

-

-

-

-

-

-

-

-

-

14

-

Count Apps
Brightfield
The Brightfield app is used for quick, label-free total cell or particle counts. A brightfield count does notdetermine cell viability. This app will count cells with a bright, white center and a defined border. The
application is not designed to count dark, solid objects.

Trypan Blue Tissue Culture
Trypan blue is a cell membrane impermeable dye that only enters cells with compromised membranes.
Upon entry, the dye binds to intracellular proteins resulting in bluish colored cells. The trypan blue
exclusion assay enables a direct counting of live (unstained) and dead (stained) cells.
Published literature suggests that using trypan blue for samples with viabilities less than 70% may result inhigher reported cell viability values when compared to fluorescent-based detection methods.
% Viability = (Live Cell Count / (Live Cell Count + Dead Cell Count)) *100.

Trypan Blue Primary Cells
The Primary Cell Trypan Blue app is configured in a similar manner as the Trypan Blue app. The
application includes optimized protocol settings for counts and viability assessment of Trypan Blue stained
primary cells such as PBMCs. For samples with high levels of debris, live/dead determination by afluorescence-based assay is recommended.

Trypan Blue Nuclei
The Nuclei Trypan Blue app uses the colorimetric membrane impermeable dye trypan blue to distinguishunlysed or intact cells from isolated nuclei or dead cells. The trypan blue dye is able to permeate themembranes of isolated nuclei, resulting in a bluish colored stain and allowing their correct identification.
Trypan is not able to enter the membranes of living cells which can then be counted as unlysed or intact
cells. For samples with high levels of debris, nuclei counting and extraction efficiency by fluorescencebased assay is recommended.
Extraction Efficiency (%) = (Stained count / (Stained + Unstained count)) *100

Erythrosin B Tissue Culture
Erythrosin B is a biosafe and non-toxic colorimetric dye that can be used as an alternative to Trypan Bluefor cell counting and viability assessment. Evaluation of cell viability using Erythrosin B dye exclusion isbased on a similar principle to Trypan Blue, that intact cell membranes exclude the polar molecules
whereas compromised membranes do not. Live cells remain unstained while dead cells are stained andhave a darker appearance.
% Viability = (Live Cell Count / (Live Cell Count + Dead Cell Count)) *100.

Erythrosin B Primary Cells
The Erythrosin B Primary Cells app is configured in a similar manner as the Erythrosin B Tissue Culture
app. The application includes optimized protocol settings for counts and viability assessment of Erythrosin
B stained primary cells such as PBMCs. For samples with high levels of debris, live/dead determination by
a fluorescence-based assay is recommended.

15

Erythrosin B Nuclei
The Erythrosin B Nuclei app uses Erythrosin B dye to distinguish unlysed or intact cells from isolated
nuclei or dead cells. Erythrosin B is able to permeate the membranes of isolated nuclei, resulting in a pinkcolored stain and allowing their correct identification. The stain is not able to enter the membranes of living
cells, which can then be counted as unlysed or intact cells. For samples with high levels of debris, nuclei
counting and extraction efficiency by fluorescence-based assay is recommended.
Extraction Efficiency (%) = (Stained count / (Stained + Unstained count)) *100

AO/PI
The AO/PI App uses green fluorescent images to capture live nucleated cell counts and red fluorescentimages for counting dead cells. The application is suited for counting cell culture samples or primary
samples.
Acridine orange (AO) and propidium iodine (PI) are both cell permeable nuclear dyes that in combination
are used to count live and dead cells within a cell solution. The AO enters both live and dead cells andresults in green fluorescing cells. PI enters only dead, nucleated cells with compromised membranes togenerate red fluorescence. Cells stained with both AO and PI fluoresce red due to fluorescence resonanceenergy transfer (FRET).
% Viability = (AO count / (AO count + PI count)) *100

AO
The AO app uses green fluorescence images to count live nucleated cells.
Acridine orange is a nuclear staining cationic fluorescent dye that enters live and dead cells and interacts
with DNA and RNA by intercalation or electrostatic attractions. When bound to DNA, it is spectrally very
similar to fluorescein, with an excitation maximum at 502 nm and an emission maximum at 525 nm.
Although AO enters dead cells, the intensity of the green fluorescence is very low for these cells comparedto the green fluoresce emitted from live cells. This app may be useful when counting solutions containing
either red blood cells and/or significant cellular debris.
The % cell viability is not provided when using this app.

PI
The PI app uses brightfield images to capture live cell counts and red fluorescence images for counting
dead cells.
Propidium iodide (PI) is a red-fluorescent nuclear and chromosome stain that only enters dead cells. Upon
entry, the PI stain binds to DNA by intercalating between the bases with little or no sequence preference.
In aqueous solution, the dye has excitation/emission maxima of 493/636 nm. When bound, theexcitation/emission maxima shift to 535/617 nm respectively.
Calculation:% Viability = (Brightfield count / (Brightfield count + PI count)) *100.

Yeast
The Yeast App uses green fluorescent images to capture live yeast cell counts and red fluorescent images
for counting dead yeast cells.
Yeast cell live/dead staining is performed using fluorescein diacetate (FDA) and propidium iodide (PI).
FDA freely moves in and out of the plasma membrane of diverse organisms from bacteria to mammaliancells. The non-fluorescent FDA taken up by cells is converted the into the green fluorescent metabolite
16fluorescein. The green fluorescence is used as an indicator of live cells. PI enters only nucleated cells withcompromised membranes to generate red fluorescence indicating dead cells.
Note: Yeast cells are smaller than typical mammalian cells and may take a little longer to settle into
position.
% Viability = (FDA count / (FDA count+ PI count)) *100

Yeast AO/PI
The Yeast AO/PI app uses green fluorescence to capture images of live yeast cells and red fluorescenceto capture images of dead yeast cells.
Acridine orange (AO) and propidium iodide (PI) are cell permeable nuclear dyes that in combination areused to count live and dead cells and determine the viability of a cell sample. The AO enters both
uncompromised intact live cells and fluoresces green. PI is only able to permeate the membrane of deadcells, quenches the AO fluorescent signal, and ultimately fluoresces red.
% Viability = (AO count / (AO count +PI count)) *100

Yeast Brightfield
The Yeast Brightfield app is used for quick, label-free total yeast cell counts. This count does not provide
sample viability information. This app will count yeast cells with a bright, white center and defined cell
membrane. This application is not designed to count dark, solid objects.

GFP
The GFP App uses green fluorescent images to count GFP transfected cells and brightfield images tocount non-transfected cells.
As GFP intensity can be highly variable it is recommended that the fluorescent threshold is set for eachsample after the count using the intensity slider in the results screen.
The green fluorescent protein (GFP) isolated from the Aequorea victoria jellyfish is a usefulreporter molecule for monitoring gene expression. GFP emits green light (lambda max = 509 nm) when
excited with UV or blue light resulting in cells that fluoresce green.
The GFP app is used to determine transfection efficiency of transient transfections.
% Efficiency = (GFP+ count / (GFP+ count+ Brightfield count)) *100

Hepatocytes
The Hepatocytes app uses a machine-learning developed algorithm, along with AO/PI, to analyze samples
in both brightfield and dual fluorescence channels. Live hepatocytes, live lymphocytes, and othernucleated cells are detected in the green channel. Dead hepatocytes, free nuclei, and other deadnucleated cells are detected in the red channel. Extra cellular debris is detected using the brightfieldchannel.
This app requires an activation code as well as a CellDrop instrument with sufficient processor RAM.
Contact DeNovix or your DeNovix Distributor for details.
Hepatocyte Viability % = (Live Hepatocytes/(Live Hepatocytes + Dead Hepatocytes))*100
Live Hepatocyte % = (Live hepatocytes/(Live+Dead Hepatocytes+Nuclei+Lymphocytes))*10017

Primary Cell AO/PI
The Primary Cell AO/PI app is configured in the same manner as the AO/PI app. The application includes
optimized protocol settings for counts and viability assessment of AO/PI stained primary cells such as
PBMCs.

Nuclei AO/PI
The Nuclei AO/PI app uses red fluorescence to capture images of isolated nuclei and green fluorescenceto capture images of unlysed or intact cells. Acridine orange (AO) and propidium iodide (PI) are cellpermeable nuclear dyes that in combination are used to count live and dead cells or nuclei within a cellsample. The AO enters both unlysed cells and isolated nuclei and fluoresces green. PI is only able topermeate the membrane of dead cells or isolated nuclei and once inside, fluoresces red. The nuclei arestained with both AO and PI but overwhelmingly fluoresce red due to fluorescence resonance energy
transfer (FRET) from the AO molecule to PI.
Extraction Efficiency (%) = (PI count / (AO+PI count)) *100

Fixed Cells PI
The Fixed Cells PI app uses red fluorescence to capture images of fixed cells. PI is a cell permeable
nucleic acid dye that is only able to permeate the membrane of dead cells. Dead cells will fluoresce red,and any debris in the sample will not fluoresce. The CellDrop’s proprietary counting algorithm is able todetermine between clustered and non-clustered cells, providing the researcher with the concentration of
fixed cells and the degree of clustering present in a sample prior to downstream sequencing preparation.

Fixed Nuclei PI
The Fixed Nuclei PI app uses red fluorescence to capture images of fixed nuclei. PI is a cell & nuclei
permeable nucleic acid dye that will permeate the membrane of isolated nuclei. Isolated nuclei will
fluoresce red, and any debris in the sample will not fluoresce. The CellDrop’s proprietary counting
algorithm is able to determine between clustered and non-clustered nuclei, providing the researcher withthe concentration of fixed nuclei and the degree of clustering present in a sample prior to downstream
sequencing preparation.

Custom Methods
The Custom Methods app allows users to count cells using brightfield, green fluorescence or red
fluorescence techniques. The application is intended to provide users with the tools to conduct
experiments using any of the available measurement channels.

18

6. Count Tab View
From the Count tab, users can preview the live image, input sample information, select the protocol forcounting, adjust the image and initiate the cell count.

Sample Name and Sample Information•

Tapping the Sample Name field will bring up the Sample Information dialogue. An optional
sample name, short description and a passage number may be entered. This information issaved with the count result.

Protocol Selector•

Protocols allow the user to adjust and save count settings within each application. Count settings
are the screening criteria used by the counting algorithms such as diameter, roundness andthreshold. See Section 7 - Count Settings for details•

Each app opens to the Default Protocol, suitable for counting most mammalian cell lines within
this application. Users may set a custom protocol as their default in the User Accounts app.

Image Controls•

Channel Selector: For the CellDrop FL model, use the Channel Selector to switch between
brightfield and fluorescence preview channels. Within fluorescence apps, the icon indicates theactive channel. Tap the icon to select a channel. CellDrop BF units and brightfield applications(Brightfield, Trypan Blue) only include the brightfield channel.

Brightfield

Green Fluorescence

Red Fluorescence

•

Focus
: The focus controls may be used when previewing any Count screen channel(brightfield, green or red fluorescence). Changing the focus when viewing one channel willautomatically change the focus of the other channels. Use the arrows to change the focus.
o Manual focus: Adjust focus while viewing the brightfield channel. Live cells should have abright center and a dark outline. The image below on the left is correctly focused. The image
on the right is out of focus.

19o For the AO/PI, AO, Primary AO/PI and Yeast Apps – check the focus in the green channel
prior to counting. In some cases, the ideal fluorescence focal plane is different than thebrightfield focal plane. A sharp focus in the green channel will
deliver the best results.
o Auto focus: Autofocus finds the best focal position for a count.
Load cells and ensure cells are roughly in focus.
Press the “Auto” button.
If auto focus does not regularly find a best focal position, select“Reset Auto Focus” from the overflow menu. Follow the
onscreen instructions. Load cells, roughly set the focus andrerun the algorithm.
•

Exposure : Touch the exposure icon to display the exposure
control in the Trypan app or for fluorescence channels. The
control is not required for brightfield counts which use a preset value.
o Trypan Blue and Primary Trypan apps: Choose the correct exposure based on theconcentration of trypan dye used or set a Custom Exposure.
▪

If the assay is 0.4% trypan dye mixed 1/1 with cell suspension, select Normal▪

If the assay is 0.2% trypan dye mixed 1/1 with cell suspension, select Low▪

Live cells should have a white center with a dark ring like the image below on the left.
The middle image is overexposed and the right image is underexposed.

▪

Custom Exposure – Some dyes used in brightfield applications have a different
opacity than common grades of Trypan Blue. The Custom Exposure Wizard instructs
the User on the steps to follow to establish proper exposure values for the 50, 100
and 400 micron chamber heights. Follow the on-screen instructions.

o Fluorescence Apps: Select a fluorescence channel. Touch the exposure icon
to open the
exposure settings. Tap the arrows to adjust. The fluorescence signal should not extend outsideof the boundaries of the cell when viewed in brightfield. The image on the left is correct. The
middle image is overexposed and the right image is underexposed. Adjust the exposure foreach channel independently.

20

•

Zoom Controls: All preview and result images on the CellDrop support zoom and pancapabilities. Pinch and zoom using two fingers to zoom in or out. Tap and drag within the image
to move the preview location.

•

Image map: A small position map in the top right is an indicator of what portion of the full image
is currently displayed on the screen.

•

Expand/Collapse: Tapping the expand or collapse icon for a preview or result will toggle
between a full screen view and the standard view.

Dilution Calculator
The dilution calculator enables the user to define target cell number or density and culture volume. The
counted result will automatically provide instructions to correctly dilute cultures to reach the desired target.
•

Tap the calculator icon to set dilution guidance parameters for the next sample count.

•

Setting targets is optional for any specific count. Inputs may be added or modified post
measurement from the Results tab Calculator screen.

•Once targets are set, they will stay in effect for all subsequent sample measurements within asession until changed.

•

When applied, the inputs are used to generate the dilution guidance presented under the Single
Sample view as well as the Results tab Results List view when the Show Dilution Guidance box
is checked.

Count Button•

The Count button initiates a measurement.

•

The required volume of sample is displayed on the Count button.

21

7. Count Settings
Each Count app includes an applicable group of count settings saved within protocols. Countsettings may be adjusted prior to a count. A subset of the parameters can be also adjusted after acount to optimize a result.
•

Cell Outlineo Enhanced Outline - Draws an outline around the identified cell boundary(recommended).
o Circle Outline - Marks each counted cell with a circle. The outlined image will berendered more quickly.

•

Sample Modeo DirectPipette - This is the default mode and is used when samples are loaded directly
into the CellDrop sample chamber.
o Slide - Choose this option when applications require cells to be contained in a slide while
counting.
•

Chamber Height: A unique feature of CellDrop Cell Counters is an adjustable chamberheight that allows for a greater dynamic range than any slide-based counter on themarket. The default chamber height is 100 µm and is recommended for most counts. Forvery low density samples, choose 400 µm. For very dense samples, select 50 µm. The
tables below includes guidelines.

Standard Magnification (FLi & BF)
Gap Height(um)Volume(uL)

Minimum Density(cells/mL)

Maximum Density(cells/mL)

400

40

7.0E+02

3.1E+06100

10

2.9E+03

1.3E+0750

55.9E+03

2.5E+07

Gap Height(um)Volume(uL)

Minimum Density(cells/mL)

Maximum Density(cells/mL)

400

40

4.3E+03

2.6E+07100

10

1.7E+04

1.0E+0850

53.4E+04

2.1E+08

Higher Magnification (FLxi & BFx)•

Small Cell Mode: Small Cell Mode can improve brightfield channel counts of small cells
near the 4 um limit of detection. The additional algorithm complexity increases the length of
the analysis.

•

Irregular Cell Mode: Enabling irregular cell mode improves counts of cells that are elliptical,
elongated or irregular in shape.
•

Dilution Factor: This value is applied to the cell count when calculating cells/mL toaccount for any dilutions or reagents added to the sample. The Dilution Factor is the ratio22

of the total assay volume to the volume of cell suspension in the assay. Enter the proper
dilution factor to calculate the correct cell density.
Example Dilution Factor calculations:o For pure cell suspensions, the Protocol Dilution Factor = 1.
o For a mixture of 5 µL of assay working solution to 5 µL of cell suspension, set Dilution
Factor = 2.
o For a mixture of 8 µL of assay working solution to 2 µL of cell suspension, set Dilution
Factor = 5.

•

Diameter (um): The Diameter settings specify the size range of objects to seek during acount. The algorithm rejects and does not process objects below the minimum size or above
the maximum limits. The limits define the boundaries of the data set, graphs andhistograms. The total allowable range is 2-400 um.
o Minimum Diameter - This setting defines the diameter of the smallest objects to be
counted. The proper value ensures that cellular debris will not be counted. Increase thissetting when counting larger cells if they are being counted as multiple smaller cells.
Decrease if small cells are not being counted.
o Maximum Diameter - Objects above the maximum diameter will not be counted. Adjust
the maximum diameter to provide a realistic size band to analyze during counts. Ifclusters of cells are being counted as a large individual cell, reducing the maximum
diameter to reflect the true cell size will improve the results.

•

Roundness: CellDrop uses Roundness settings to identify cells based on morphology. The
higher the roundness, the more circular and completely outlined a cell must be to be
included in the count. Reducing the roundness setting allows cells that are not perfectly
circular or cells with incomplete structures to be counted. If single cells are counted as
multiple cells or artifacts are counted, increase the roundness setting. If cells are notcounted, decrease the roundness.
o Brightfield Roundness - Used for Brightfield counts.
o Live Roundness - Used in apps counting live cells.
o Dead Roundness - Used in apps counting dead cells.
o Green Roundness - Used in the Custom Methods app for the green fluorescencechannel.
o Red Roundness - Used in the Custom Methods app for the red fluorescence channel.
o FL Roundness - Used in the GFP app to count fluorescing cells.

•

Threshold: The CellDrop Series uses Threshold settings to evaluate each counted cell to
see if it is above the intensity criteria set in the protocol. The higher the threshold is, thegreater the intensity must be to be included in the count.
o Stained Threshold - Used in the Trypan Blue app to determine if a cell is dead or alive.
The higher the setting, the darker a cell must be to be counted as dead. If dead cells are
not being counted, decrease the threshold. If live cells are being counted as dead,increase the threshold value.
o Green Fluorescence Threshold - Used in apps that count cells using the green
fluorescence channel.
o Red Fluorescence Threshold - Used in apps that count cells using the red fluorescencechannel.

23

o Decrease the fluorescence threshold if cells that are positively stained are not being
counted in the analysis and vice versa.

24

8. Results Tab View
This section details the Results Tab user interface and some differences seen when an instrument is inStandard Mode or 21 CFR Mode.

STANDARD MODE VIEW

Export Icon
The export

button is active immediately after a count in Standard Mode.

Next Count
This button changes the view back to the Count tab and starts a live preview. The previous result is not
saved until the user raises the arm. If the Count button is pressed without raising the arm, the previous
count will be overwritten. To capture a new result of a new sample, raise the arm, clean the upper andlower sample surfaces, lower the arm then load the next sample.

Common controls
All remaining controls (i.e. Optimize Settings) are available in both Standard and 21 CFR mode andare detailed in the next section of the guide.

25

21 CFR MODE VIEW

Export Icon
The export

button is inactive until the record is electronically signed.

Discard. New Count
This button is visible when the result is un-signed. Tap this button to discard the result and return theuser to the live preview in the Count tab. The result is not saved. SOPs should be established related
to the use of discard to avoid ‘testing into compliance’ scenarios. The Discard button may be disabled
by an Administrator account using the Permissions tab in the Settings app.

Save and e-Sign
The user may optimize settings at this time. Once satisfied with the result, the user electronically signs
the record and saves the result using the Save and e-Sign button. A result is not saved and may
not be exported until the result is electronically signed. The electronic signature dialogue willopen. The logged in user may sign a record with any valid Signature Meaning for his/her User ID. The
password is also required for the first signing of a session. 21 CFR Mode configuration details arefound in the 21 CFR section of this guide.

26

Export and Backup of Signed Record
After signing a record, the user will be asked to export the signed record to a location of the user’s
choosing. The choice will be saved during the active Count app session.

Exporting the record ensures that the valid electronically signed record as a human readable pdf, as
well as any associated files selected, are exported to a secondary location. This creates a backup of
the record which should then be further managed according to the organization’s policies.
IMPORTANT – Organizations should establish SOPs to ensure that signed records from the
instrument are exported and backed up on a routine basis. System Administrators can search forresults in the Data App that have not been exported.
Records that have not been exported are not backed up and are subject to loss. The end user’sorganization is solely responsible for ensuring proper procedures are followed for electronic record
backup.

Controls After e-Signing Record
After signing a record, the following buttons are displayed:27

Signed. Next Count
This button indicates the record is electronically signed and saved. Pressing the button will take theuser back to a live preview on the Count Tab. The data is saved. Pay-As-You-Go systems deduct a
count after the arm is raised to remove the previous sample.
Add SignatureThis button allows the logged in User to add a signature to a previously signed record.
Record Security
Electronically signed results may not be altered. If the user chooses to optimize settings, change
gating or in other ways alter the result, the buttons revert to Discard. New Count and Save and eSign. The user may electronically sign and save the new result. The new result is saved while theoriginal record is unaltered and is retained.

28

9. Result Tab Common Controls

Optimize Settings•Once a sample has been counted, a subset of the protocol parameters can be further
adjusted to optimize the result. Press the Optimize Settings button, adjust settings and thenpress Count with Current Settings to reanalyze the existing image(s) using the new
settings. Users have the option to save the adjusted settings to a protocol if desired by using
the Save to Protocol button that appears when a change is made.

Cell Size Gating•

Use the on-screen sliders to adjust the size range of interest for the current sample. Celloutlines and count results are updated in real time. Gate to exclude debris or alternative cell
populations from analysis. Press the Histogram button to graphically view cell size
distribution and apply gating. Press Apply Gating to Result to update and save the result.

Intensity Gating (GFP app)•

In the GFP app, users may gate cells by size as well as intensity. Use the on-screen slider toadjust the intensity gating from 0-100. The higher the number, the higher the intensity mustbe for a cell to be included in the counted result. The intensity gating icon will open a graphshowing the intensity profile of the cell population.

Result Overlay Selector

Brightfield

Brightfield/
Green Overlay

Green
Fluorescence

Brightfield/
Red Overlay

Red
Fluorescence

Green/ Red
Overlay

Brightfield/
Green/Red
Overlay29

Action Bar Controls•

View Selector: Use this drop down to access the various Results tab views including viewing
a list of multiple results. The Default selection is Single Sample view.

•

Outlines: The checkbox is used to toggle on/off the outlines for counted cells for views that
include cell images.

•

Export: Export results in .csv, .pdf, or .png formats via USB, Email, Network Folder andNetwork Printer.
21 CFR Mode: Only results that have been electronically signed may be exported.

•

Image Selector: This icon indicates the overlaid images included in the result being viewed.
Tap the icon to change the overlay combination for the current sample or after optimizing
settings for a previous result.

•Calculator: Access the Dilution Calculator.

Additional Action Bar Controls•

Select/Deselect All
one tap of the icon.

: These icons enable all items in a list to be selected or deselected with•Edit
: Highlight a single sample and then tap the edit icon to add or modify sample
information. Changes will overwrite the information stored in the database and will be appliedin all lists referencing the sample.
21 CFR Mode: Record may not be edited from this view.

•

Delete : Tapping this icon will delete the highlighted samples from all results lists.
21 CFR Mode: Signed records may not be deleted. Results can only be removed via
archiving by a System Administrator. See 21 CFR section for details.

Results View Options•

Single Sample
o Sample Number: The arrows on either side of the sample number are used to switch
between samples measured in the current session.
o Sample Name: This is an editable field. Touching will open the Sample Information
dialogue for input. Changes will overwrite the information stored in the database and will beapplied to all lists referencing the sample.
21 CFR Mode: Sample information may be edited prior to signing a result. To edit and savethe sample information for a previously signed result, a new signature is required.
o Results Panel: A table of results will be displayed in the Results panel. If dilution
parameters were provided at the time the count was initiated, then dilution guidance will
also be presented in the panel.

•

Results Listo The list shows all samples counted in the current measurement session.
o When the Show Dilution Guidance box is checked, the dilution guidance is displayed witheach result in the list for which dilution information was entered.

30

•Image Listo The list displays all samples counted in the current measurement session.
o Cell Count: The sample cell count is presented in the table. This value automatically
updates whenever the results are recalculated due to a change in cell size limits or othercounting parameters.

•

Graph
o Includes a summary of results to allow the user to select different charts to add.
o A color legend is included for the various histograms. Select up to 5 samples at a time tochart.
o Graph Type Menu: Use the drop down to select additional graph types.

Graph Types•

The graph types available are app dependent. Graphs may be expanded to full screen foreasier viewing using the expand/collapse button.

Graph showing totals only or with a cell size distribution histogram•

Total Cell Count: Available for the Brightfield and GFP apps only.

•

Live Cell Count: Available for all apps except Brightfield.

•

Dead Cell Count: Available for Trypan Blue, PI, AO/PI and Yeast apps.

•

Live and Dead Cell Count: Available for Trypan Blue, PI, AO/PI and Yeast apps.

•

GFP+ Cell Count: Available for the GFP app only.

Additional Views•

Cluster #: Available for all apps. Displays count of cell cluster sizes.

•% Viability: Available for Trypan Blue, PI, AO/PI and Yeast apps.

31

10.

Export and Print Options
There are several export and print options available on the CellDrop. Data export options are accessedusing the export iconin the blue action bar. The screen capture option is accessed from the overflow
menu. Data available to export includes results, protocol settings, images, and histograms.

Screen Capture
The screen capture feature in the overflow menu is used to export a .png file. Screen capture is available
for most EasyApps screens.

ExportTapping the Export icon will bring up a series of two dialogue boxes. The first dialogue is used to selectthe desired combination of results, images and histograms to include in the export. The second dialogue isused to select the export destination.

Export Selections•

CSV: The Results and Protocol settings will be presented in a .csv format that can be opened in
Excel or uploaded to a LIMS system. Additional sample related information not displayed in the
Results tab Single Sample view is included when exporting in a .csv format. To open an
exported .csv file on a computer, the DeNovix instrument and the computer must both be set touse either period or comma number formats. The CellDrop number format is determined by thelanguage selected using the Settings app.

•

PDF: A report .pdf file will be generated based on the selected options during export. Report
options include Results and Protocol Settings, Images and Histograms.

•

IMAGES: CellDrop images are exported as .png files. Channel specific images and the current
result image may be exported. The available options are App dependent.

•

The result image for the active result can be modified to show the channels desired in theoverlaid image. The channel selector button shows the combinations available. Cell outlines may
also be toggled off if desired. To change the overlaid image of a past result, reprocess theimage using the Optimize Settings button. Then select the new overlay desired using thechannel selector.

•

Graphs: Use Screen Capture from the Overflow menu to capture and export histograms or bar
charts found in the Graph screens.

Export Destinations•

Export destinations include: USB, Email, Network Folders and Network Printer. See the Settings
app section for Email, Network Folder and Network Printer set up.

32

11.

Utility Apps

User Accounts
Standard Mode

This section describes the User Accounts app in Standard Mode. See the next section for 21 CFR
Mode.
The User Accounts app allows users to create and manage individual accounts on CellDrop. Users
can adjust their default settings. Accounts are also used as a means of segregating data when
performing searches in the Data app. For Pay-As-You-Go models, users may also define how countcodes are shared amongst user groups.
App Structure
Accounts are listed in the left pane of the screen. A menu of app function options is presented in theaction bar, where the selected option is highlighted with a blue background.
•Select a user account from the dropdown list in the top grey bar and select an option to edit:
Profile, Report Emails, File Share, and Defaults.

App Functions
There are five basic functions that are controlled using the Account app.
•

Adding and deleting accounts•

Managing passwords for accounts•

Managing email addresses associated with an account•

Setting account specific app protocol preferences

Adding or Deleting Accounts
There are two preconfigured system accounts built into the software - the General Account and thePrimary Administrator Account. These accounts may not be deleted.
Primary Administrator•

This account is used to manage instrument settings. It is not used within the countapplications.

•

The primary administrator account is not initially password protected. A password may be
applied at any time.

General Account•This account is used as the default selection for all apps each time an app is opened. Thisaccount may not be password protected or deleted.

There are two types of accounts that may be added, edited or deleted as described below.
User Added Standard Accounts•

Any user may add a standard level account. It is not necessary to select an account before
adding a new standard account.

•Standard accounts may only modify or delete protocols or emails created from within theiraccount.

User Added Administrator Accounts•

An administrator level account may only be created by another administrator. The primaryadministrator or another existing administrator account must be highlighted in order to enable
the adding of a new administrator.

33

•Administrator accounts may modify or delete user entries of any sort created from within any
account. A drop-down menu will be used to select the account of interest.

•

Administrators may change the password for any account as well as delete any account.

Account Passwords•

Passwords are optional for all added administrator or standard user accounts.

•

Contact the holder of an administrator account if a standard user account password isforgotten.

•

Contact DeNovix customer support or your local distributor if the Primary Administrator
password is forgotten. Please have the serial number of the instrument available whenrequesting help for the Primary Administrator password reset.

Deleting an Account•When an account is deleted, all protocols and custom report formats created by that accountare reassigned to the General Account.

•

The account specific saved data will also be permanently removed from the instrument at thetime the account is deleted.

Tab - Email Addresses
Multiple “to” email addresses for exporting results may be set up for each account. The nickname isused in the drop-down list presented on an export destination dialog box.

Tab - Defaults
Set the default protocol you would like to have at start up for this user account. Select the app andthen choose any established protocol as the default.

User
Accounts
21 CFR Mode

This section describes the User Accounts app in 21 CFR Mode. See the previous section for Standard
Mode. Account management features must follow rigid guidelines in 21 CFR mode.

Account Management for 21 CFR Part 11 Compliance
System Administrators use the User Accounts app to create and manage user accounts on the device.
The app is also used for selecting roles, deactivating users, setting temporary passwords andchanging account information. Each user is required to have a unique User ID and password thatmeets the organization’s password rules set in the 21 CFR App. All User Account changes arecaptured in the 21 CFR Audit Trail data.
Accounts also serve as a convenient means of segregating data when performing searches. For PayAs-You-Go systems, users may also define how count codes are shared amongst user groups.
IMPORTANT: Organizations are responsible for following the strict security requirements of 21
CFR Part 11 regulations. DeNovix software enables a method of compliance, but organizations
are responsible for establishing and following procedures to ensure compliance.

App Structure
User accounts are listed in the left pane of the screen. Within a user’s own account, tabs in the rightpane allow access to User Profile, Emails, Network Folders and Default settings.

34

Administrators can access the Profile tab of other users by selecting that user from the list in the left
pane.

App Functions
Administrators have the following functions within the User Accounts app.
•

Add accounts•

Set Account Type•

Assign Role•

Change first name or last name•Set Temporary Passwords•

Deactivate / Reactivate accounts

Adding Accounts
Choose the + symbol from the left column to add a new user. Assign a User ID and the user’s
information. Create a temporary password for the new user. All users must have a unique User ID.

35

Account Type
When a new user is added to the system, the Administrator must define the Account Type.
Administrators can change the Account Type of any user.
•

User – This account type has full access to count apps and permitted utility apps. User
account types do not have any access to Administrator level system settings. A user may
change first name, last name or password of his/her own account, but the user cannotchange anything related to other user accounts.

•

Administrator – This user type has full access and control over all system settings. At least
one Administrator account must always be active in 21 CFR Mode. All Administrator
accounts have the same administrative controls on the device.

Role
Each user must be assigned to an available Role as defined in the 21 CFR App. See 21 CFR App
section for configuration details.

Account Passwords•

Passwords are required for all Users.

•

Password must follow the organization rules set in the 21 CFR App (special characters,
length, etc.)•

A user may not reuse a password they have previously used.

•

If a user is unsuccessful logging in over a set number of times (set in 21 CFR app), theaccount is locked. An administrator can set a temporary password to unlock the account.

•

Organizations are responsible for establishing password security procedures to ensurecompliance with 21 CFR Part 11requirements.

•

Contact DeNovix customer support or your local distributor if the sole Administrator password
is forgotten. Please have the serial number of the instrument available when requesting help
for the reset.

Deactivate / Reactivate an AccountUser accounts may be deactivated by an Administrator. Per 21 CFR Part 11 guidelines, a user account
may not be deleted. User activation/deactivation is tracked in the Audit Trail.

Tab - Email Addresses
Multiple “to” email addresses for exporting results may be set up for each account. The nickname isused in the drop-down list presented on an export destination dialog box.
Tab - Defaults
Set the default protocol you would like to have at start up for this user account. Select the app andthen choose any established protocol as the default.
DataThis app allows the user to search for all saved data.
•

Data may be filtered in the following ways:oStandard mode: App, date, protocol name sample, Account nameo

21 CFR Mode: App, export status, User ID, Signature Type, date•Once the filter criteria have been selected, tap the Search button at the bottom of the screen.
Tap any column header to sort the results list.

•

Highlight the sample(s) of interest and tap the Add to Report button to include the samples
on Results tab pages.
36•All data must be from the same app type for the samples to be added to the Report tab.

•Features and functions previously described for the Result screens apply to thecorresponding Data App Report screens.

Export•All data is automatically saved to the CellDrop onboard memory at the time of measurement
and may be exported as described in the previous section.

•

When the export function is used, the data is not deleted.

Delete21 CFR Mode•
The Delete function is not active for instruments in 21 CFR Mode. Data can only be archived
from the system by an Administrator through the 21 CFR app.
•

The storage capability of the CellDrop is a minimum of 120 GB of data. If the storage isapproaching capacity a warning message will prompt the need to free space for continued
operation.

Standard Mode•

The Delete function permanently removes the selected data from the instrument databaseand cannot be restored to the instrument.

•

The deleting privilege for standard accounts is enabled or disabled using the Settings app.
When disabled, only administrative accounts may delete or archive data.

•

The storage capability of the CellDrop is a minimum of 120 GB of data. When the storage
capacity has been reached, the software uses a first in/first out paradigm for remaining
sample data. The Delete function should be used as an infrequent method of removing alarge amount of data from the CellDrop database.

•The delete process continues to work even when the interface goes to sleep after 10 minutes
of no user activity. Touch the screen to wake-up the user interface and update the progressmessage.

Count Codes

The purpose of the Count Codes app is to manage pre-paid counts using codes on a per accountbasis. This app will appear on instruments purchased on a pay-as-you-go basis.
The Primary Administrator account is only used to add, review or transfer codes for other accounts.
This account cannot be used to make cell measurements.
For Admin accounts and Standard accounts- Tab menu options include:•

Enter New Code•

Review Code History•

Transfer Counts•

Share Counts

Administrators may transfer or share counts between all accounts on the instrument.
Standard accounts may transfer or share counts from only their account.
Please include the serial number of your instrument when contacting DeNovix by email to purchaseadditional count codes.
Outside of the US, please contact your local distributor for assistance.

37Calculator

The Calculator app includes a scientific calculator as well as convenient dilution calculators.

Timer

The Timer app enables the setting of up to two timers at one time. An alert will display when a timer
has finished its set runtime.

Settings

The Settings app may be found on the second Home screen page. Only administrator accounts may
access and change settings. The app provides access to the following functions and information:
System Info•

View software and firmware versions, IP addresses, current language and hostname.

Date/Time•

Select manual or automatic date and time options.

Language•

Select the language of operation for the device. Not all languages are fully translated. Setting
the language also determines whether a period or a comma is used for the decimal place.

Network Information•

The Wi-Fi and Ethernet MAC addresses and IP addresses can be found under this setting.

Permissions•

Use Permissions to set access to the Settings app tabs for non-Administrator accounts.

21 CFR Mode: Administrators can disable the “Discard” button from this screen if desired.

38

Sound (enabled for all models in future release)•

Play a test sound.

Storage•

Displays current storage statistics.

Wi-Fi
•

Turn Wi-Fi on or off and connect/disconnect from a network.

•

Connect CellDrop to any 2.4 GHz network. SN-07409 and higher may also connect to 5 GHz
networks.

•

Select the country where the unit is installed in order to ensure the Wi-Fi radio uses thecorrect frequency bands.

•

DeNovix instruments are compatible with eduroam using login credentials provided by yourinstitution. See Technical Note 225 on denovix.com for a step-by-step guide to connect.

Ethernet•

Ensure that the Ethernet cable is connected.

•

Tap the Edit button to configure Ethernet connection.

Contact your local IT department if additional help setting up Wi-Fi or Ethernet is needed.

Display•
Includes controls for screen brightness and when the screen sleeps.
Network Printing•
Search and add compatible network printers for easy export of hard copy reports.
Email
Email Set-up•

It is recommended that a free, dedicated Gmail or Outlook account be used for outgoingemail from the instrument. Follow instructions to create a two factor authentication (2FA)“app passwords” for the account in order to ensure emails can use current SMTP and TLS
protocols.

•

Ensure that the instrument is connected to Wi-Fi or Ethernet.

•

Launch the Settings app and navigate to the Email section on the left scroll menu.

•

Enable the POP3 option on your Email configuration. Do not use an IMAP account as
messages from IMAP accounts could occupy a significant amount of the instrument's
memory.

See Technical Note 205 on denovix.com for a step-by-step walkthrough of email setup.
Refer to your IT support group and CellDrop Technical Notes on denovix.com if additional assistanceis required.

39

Network Folders
The instrument can export to SMB network folders connected to the same network as the instrument.
For best results connect to the network through an ethernet connection, especially if exporting pdfs
and images.

Enter the following information. See DeNovix Tech Note 122 or your IT Department for additional help.
•

Name - Create a name for your network connection.

•

Server - The IP address of computer or server where the shared folder is located. The IP
can be found in Windows Network Status under “Properties” or by entering ipconfig at theWindows command prompt.

•

Domain name - The domain name when the shared folder resides. From a windows
command prompt enter echo %userdomain%. Do not include slashes or backslashes.

•

Share path - The share name of the shared folder. In Windows, find this by right clicking on
the shared folder and viewing the properties. The share name should be a single word. No
slashes or backslashes.

The example below shows a correctly shared folder.

•

Mount point (optional) -The path to a subfolder within the main shared folder. Example
input sub1/sub2 is sub1 is a folder inside the main shared folder and sub2 is within folder
sub1.

•

User name - The username accessing the share. No slashes or backslashes.

•

Password - Password of the User name account.

Press Connect to connect to the network folder. Press Send Test File to transfer a time stamped
text file denovix.txt to the network folder.

40

UpdaterThis app can be opened by Administrator level user accounts. Updater is used to update CellDrop
software and firmware to the most current available versions. Updates may be downloaded using theUpdater app when connected to Ethernet or Wi-Fi. An Ethernet connection is recommended forsoftware updates.
Updates maybe downloaded on line or downloaded to a FAT32 formatted USB drive fromdenovix.com.

ScreenLocker
Launch the ScreenLocker app from the Home screen prior to cleaning the touch screen. This will
disable swiping motions from activating other software activities. Tap the home button to exit.

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12.

21 CFR App

21 CFR

DeNovix 21 CFR Part 11 Compliance Ready Software is available for activation on CellDrop
instruments. An activation code is required and can be purchased from DeNovix or a DeNovix
distributor. Once activated, 21 CFR Mode provides a suite of seamless, integrated controls forelectronically signing records, managing user accounts, configuring the organization and accessing theaudit trail. The software enables the following:•

User login with unique User ID and password for each user.

•

Operation as a closed system with a single, continuous period of controlled system access.

•

Users review results and electronically sign records while logged in. Records can beelectronically signed with a Signature Meaning that is associated with the user’s Role.

•

Automatic export (back up) of electronically signed records to a second location whensigning.

•

Export of human readable pdf reports and optional raw data such as images and csv files.
The pdf report includes details of each electronic signature associated with the record.

•

A user interface for viewing stored information but no ability to directly access and alterelectronically stored data.

•

Administrator tools to customize and manage the organization’s Roles, Signature Levels andSignature Meanings.

•

Administrator tools for User Account management including Role assignment, activation,
deactivation, name changes and password resets.

•

An Audit Trail function to automatically log system events and user activities in a database. If
audited, System Administrators can easily search the Audit Trail and export a humanreadable .csv report as desired.

•

A convenient configuration wizard for easy set up and customization during activation.

Activation Wizard Overview
The activation wizard is a step-by-step procedure for setting up CellDrop to comply with yourorganization’s needs. Default valves are included but are fully configurable during activation. 21 CFR
Mode:•Each user is assigned to Role•

Each Role has a Signature Level which establishes a rank in the organization•

A set of Signature Meanings must be defined (tested, rejected, approved, etc.)•

Roles are linked to one or more Signature Meanings, meaning that any User who is assigned
to a Role can electronically sign a record with one of the available signature meanings.

•

Once the organization rules are configured, each user is assigned to a Role. This establishes
the signature privileges of each user.

Activation Wizard 1/7 - Administrator AccountTo begin, open the 21 CFR app and select an Administrator level account from the Account selector at
the top of the screen. If a user created Administrator account isn’t selected or none exists, the user is
prompted to create a new Administrator account.
When 21 CFR Mode is activated, the open access accounts “General” and “Primary Admin” aredeactivated. For PAYG systems, any existing count balance in the General account is automatically42

transferred to the Administrator activating 21 CFR Mode. The Administrator can use the Count Codeapp to redistribute counts.

Activation Wizard 2/7 – Enter Activation CodeEnter the twenty-digit activation code and press Apply Code.

Activation Wizard 3/7 - Roles
Users will be assigned to a Role in the final step of activation. In this step set up the Roles for theorganization. Each Role includes a value for Signature level. The Signature Level value establishes ahierarchy in the organization. Users can view data for users at their Signature Level and below. But
they cannot view data for Roles with higher Signature Level number.

Activation Wizard 4/7 – Signature Meaning
Each User, through their assigned Role, will be able to electronically sign records with the Signature
Meanings as defined in this table. Configure the Signature Meanings as required for the organization
but using the add, edit and delete icons. At least one Signature Meaning must exist.

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Activation Wizard 5/7 – Role-Signature Meaning Linking
In this step, set the Signature Meanings for each Role. Select a Role from the table and then check
Include to allow that Role to electronically sign records with that particular Signature Meaning. In theexample shown, the Sr. Technician Role can e-Sign records with Tested, Reviewed and Edit sample
name/description Signature Meanings. Any Role must be linked to at least one Signature Meaning.

Activation Wizard 6/7 – Password Rules
Set password rules as desired for the organization. Some requirements are optional. Minimum
password length and account locking after failed attempts to login are requirements of 21 CFR Part 11
compliant software.

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Activation Wizard 7/7 – Existing Users
The final step of the Activation Wizard is to complete the user records for any existing user accounts
that are on the system. Each user must have a User ID, First Name, Last Name, Role and atemporary password set in order to complete the wizard. Edit user accounts by selecting them fromthe list and then using the edit icon.
Note: This is last opportunity to delete existing user accounts. After 21 CFR activation is completed,user accounts may be deactivated, but they may not be deleted.

Audit Trail
DeNovix 21 CFR software continually creates an Audit Trail which is a time-stamped log of events andchanges that take place in the system throughout the life of the instrument. Administrators can search
the audit trail to filter the results by date, Event Type, User ID and Signature Meaning.

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Event types tracked include Signature, System, User Account, Record Management andApplication/Machine errors. Some examples of logged events are the electronic signature of a result,deactivating a user, a failed login attempt, exporting a result and archiving a record. This list is only
representative and is not a complete list of all actions logged.
Set the filters as desired and press Search to update the result list. Select records and press “Add toReport”. Move to the Report Tab to see additional information about each audit trail entry. To export a
copy of the Audit Trail as a .csv file, select records, press the export icon and select the destination.

Archiving
Results that have been previously exported from the instrument may be permanently deleted throughuse of the Archiving function in the 21 CFR app. This is the only mechanism for removing data fromthe instrument. Prior to removal, the Administrator should confirm records that will be removed havebeen properly backed up to another location. Use the Data app to export additional copies of theelectronically signed results prior to archiving. Archiving immediately and permanently deletes alldata and images associated with the signed record. Do not archive a record before confirminga secure copy of the record exists in a secondary location or has been properly managed
according to the organization’s records retention policies. All Archiving events are captured in the
Audit Trail database.

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Filter the results by date, App, User ID and Signature Type as desired. Touch Search to update thesearch results. Select desired results and then Add to Report.

The Report tab view shows if a result has been previously exported from the system. Only results thathave been exported can be selected for archiving. Touch the delete icon to initiate archiving. The
system Administrator must enter their password to complete archiving. The archived records are
immediately deleted. Since the record has been previously exported, an additional backup copy
is not created at the time of archiving.

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13.

Troubleshooting and Maintenance
Quick Help
Most issues concerning accuracy or reproducibility are sample, technique or protocol related and may
be resolved by following the suggestions below. See Technical Note 186 – CellDrop Best Practices for
additional details.
•

Follow initialization instructions when powering on unit. Do not leave any materials between
the sample surfaces. Clean with ethanol and press OK to complete the initialization.

•

Clean sample surfaces between samples using a dry, folded laboratory wipe. Wipe both
samples surfaces firmly in one direction.

•

Do not over or under fill the counting chamber. Follow the Count button dispense volume
directions. Volume is dependent on the chamber height specified in the protocol.

•

Ensure that samples are monodispersions of cells in solution. Vortex or thoroughly mix with apipette before loading into the count chamber.

•

Ensure that cells are in focus before tapping the Count button. Live cells should have a sharp
dark ring and a bright white center. Tap the (i) button in the focus adjustment control for
example images of properly and improperly focused cells.

•

Ensure the proper exposure is set for Trypan Blue and Fluorescence counts. Tap the (i)button in the focus adjustment control for example images of proper exposure. For
fluorescence applications, the exposure of the green and red channels are adjusted
independently.

•

Ensure that all sample concentrations fall within concentration limits of the instrument for
accurate results. When measuring samples with very low or very high cell densities, it is best
to use an alternative chamber height setting and sample volume.

•

Review the protocol settings used to make the sample count. Some parameters such as cellsize limits, fluorescence intensity thresholds and cell roundness values may need to be
empirically determined for the cell type and culture solution in question.

•

Ensure that cells have stopped moving before tapping the Count button. The time for cells tosettle may vary by cell type and should be determined empirically.

•

Avoid introducing air bubbles into the sample solution when mixing or dispensing samples.

•

Refer to the manufacturer of cell dyes such as acridine orange, propidium iodine or FDA for
assay specific troubleshooting guidance.

Software Errors
Pop-up messages will provide guidance for software-based errors or invalid user actions. In mostcases, simply re-launching the app will allow the user to continue to make measurements. Update thesoftware using the Updater app if experiencing a reoccurring software error.
Contact DeNovix Customer Support at 302-442-6911 or info@denovix.com if additional help is
required. Please include the serial number of your instrument and all pertinent data when contacting
Customer Support by email. The serial number may be found on the bottom of the instrument.
Outside of the US, please contact your local distributor for assistance.

Cleaning
Between Measurements:•
Use only a dry, lint-free lab wipe to clean top and bottom sample surfaces. Check the live
preview to verify the sample has been removed.
•

Optional: For additional disinfection or cleaning if sample residue has dried on the surface:
Lower the arm and pipette 15 µL of 70% ethanol or 10% bleach into the sample chamber.
Wait approximately 10 seconds. Lift the arm and wipe both the top and bottom sample
surfaces with a dry, lint free wipe.

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Full Instrument Cleaning:•
To clean the entire instrument, spray 70% ethanol or 10% bleach onto a paper towel first andwipe down the surface of the instrument.
•

Warning! Do not use a spray bottle to douse the entire instrument. Excessive liquid may
damage internal components.

Solvent Compatibility
CellDrop Series instruments are compatible with most solvents typically used in life sciencelaboratories.
Exception: Do not use Hydrofluoric Acid (HF) as the fluoride ion will etch the surfaces of themeasurement surfaces.

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14.

Customer Support
DeNovix Inc.
3411 Silverside Road, Hanby Building
Wilmington, DE USA 19810
302-442-6911
info@denovix.com
www.denovix.com
Please include the serial number of your instrument when contacting Customer Support by email. If
sending Customer Support a screen capture or a report file using the CellDrop email feature include
your name, institute/company, phone number and return email address. Please note responses will
not be sent directly to the email account used by the instrument.
Outside of the US, please contact your local distributor for assistance.

Warranty
All product(s) and accessories sold by DeNovix Inc. are under warranty against manufacturing defects.

Disclaimer
All information in this document is for reference purposes only. DeNovix Inc. makes no claims that thisdocument is complete or error-free and assumes no responsibility and will not be liable for any errors,omissions, damage or loss that might result from any use of this document.

Instructions for Disposal of Packaging and Waste Equipment
DeNovix encourages customers to locally recycle shipping and packaging materials. Corrugatedcardboard and paper can be recycled wherever it is accepted locally (e.g., curbside bins, recycling
facilities, etc.). For foam and plastics, please contact your local recycling office for information on how
to recycle.
When an instrument has reached its end of life, DeNovix recommends that customers responsibly
dispose of the equipment by following in line with local regulations. Material construction is available
upon request. Visit denovix.com/sustainability for more information.

Research Use Only
DeNovix Inc. products are for research use only.

© 2024 DeNovix Inc.

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