(Appendix)
Available Instruments○ Flow Cytometers
Flow cytometers can measure cell size, cell granularity, the amounts of cell components such as total
DNA, RNA, proteins and lipids labeled with fluorescent probes.
• FACSCanto II (Becton-Dickinson):Laser excitation source: 488 nm, 633 nm, 405 nm
Examples of detectable fluorochromes: FITC, PE, APC, Alexa Fluor (488, 647), PerCP,
Pacific Blue, Cyan, AmCyan○ Cell Sorter• FACSAria II (Becton-Dickinson):
FACSAria II is a high speed cell sorter for measuring and sorting fluorescently labelledcells. The instrument contains four lasers (488, 633, 405, and 375 nm) for excitation. The
desired cells can be sorted into a tube or a multi-well plate (6, 12, 24, 48, 96, or 384 wells).
Examples of detectable fluorochromes include FITC, PE, APC, Alexa Fluor (488, 647),
PerCP, Pacific Blue, Cyan, AMCyan, and Hoechst Blue/Red.
・ SH800 (SONY):
Laser excitation source: 488 nm, 633 nm, 405 nm
The desired cells can be sorted into a tube or a multi-well plate.
○ DNA Sequencers
These instruments can automatically determine the base sequence of DNA molecules.
• ABI (Applied Biosystems) 3130: Samples must be prepared on a 96-well plate specific to theinstrument.
○ Real-Time PCR Systems
A thermal cycler and a fluorescence spectrometer are combined into one in these systems. Thesystems can detect PCR products as they accumulate in real-time during the PCR amplification
process. The resulting amplification curve allows accurate quantitative analysis from a small amountof sample before amplification began.
•StepOnePlus (Applied Biosystems):Samples must be prepared on a 96-well plate. The optical detection system is LED-based,and detectable fluorescent dyes include FAM, SYBR Green, VIC, JOE, and ROX. Gene

expression profiling and SNP genotyping experiments can be conducted on thisinstrument.
•7300/7500 Real Time PCR System (Applied Biosystems):
Samples must be prepared on a 96-well plate. Assays using TaqMan probes or SYBRGreen can be used to quantify the gene expression level and to conduct SNP analysis.

○ Confocal Laser Scanning Microscopes
Fluorescent and transmitted lights from a sample excited by a laser beam are detected, and a two- or
three-dimensional image of the sample is obtained. Time-lapse observation of the samples isavailable with FV1000-D. The obtained images can be analyzed using accompanying software.
•FluoView FV1000-D (Olympus):•

Available laser wavelengths: 405 nm, 473 nm, 559 nm, 635 nm•

Examples of detectable fluorescent dyes: DAPI, Hoechst Blue/Red, Fura Red,
EYFP, Alexa Fluor (488, 647), Rhodamine, Cy5○ Motorized Autofocus Inverted Microscope•

IX81-ZDC-Meta (Olympus):
This microscope can be applied to a wide variety of experimental techniques from highsignal-to-noise ratio fluorescence microscopy to phase-contrast microscopy. It produces
two- or three-dimensional images of the sample, and can accurately track the sample’schanges over a long period of time. The obtained images can be analyzed using
accompanying software.

○ Cryostat Microtome•

CM3050 S (Leica)
This instrument is used to prepare frozen tissue sections for microscopic observation fromlarger, rapidly frozen specimens, such as organ sections.

・ RM2235 (Leica)
Manual Rotary Microtome for Routine Sectioning.
○ Ultracentrifuges• Optima TLX (Beckman): maximum rotational speed = 100,000 rpm• Himac CP 100MX (Hitachi): maximum rotational speed = 100,000 rpm• XPN-80 (Beckman): maximum rotational speed = 80,000 rpm○ Image Analysis Systems

These systems use highly sensitive CCD image sensors for chemiluminescence, bioluminescence,fluorescence, and/or visible imaging. The data is saved onto a computer for image analysis.
•ImageQuant LAS 4000 Mini (GE Healthcare):
Imaging applications: chemiluminescence, bioluminescence, fluorescence, visible light○ Microplate Reader•

POWERSCAN4 (DS Pharma Biomedical)
This microplate reader can accommodate 1- to 1536-well microplates, and can measureabsorbance, fluorescence intensity, luminescence, time-resolved fluorescence, andfluorescence polarization.

・ BioTek Synergy LX (Agilent technology)
This microplate reader can accommodate 96 or 384 well microplates.
○ Imaging Systems for Small Animal Experimentation*•IVIS Spectrum (Xenogen):•This in vivo imaging system can detect and image small amounts ofchemiluminescence and fluorescence in mice and rats under anesthesia by using ahighly sensitive, cooled CCD camera.

••

Nominal pixel size: 13.5 microns•

Resolution: 20 microns•

Quantum efficiency: ≥85% for 500-700 nm wavelengths

LaTheta LCT-200 (Aloka):This instrument is an X-ray CT scanner for small animals (mice, rats, and small rabbits). It
can continuously acquire tomographic images at high speeds with a high resolution of 24
microns, and can construct three-dimensional images.
Typical scanning times (approximate values):• Tomography mode (180° rotation)•Standard mode: 5.8 s/rotation•

High precision mode: 11.6 s/rotation•

Ultra high precision mode: 23.2 s/rotation•

Integration mode: 23.2×N s/rotation• Tomography mode (360° rotation)•

Standard mode: 10.6 s/rotation•

High precision mode: 21.2 s/rotation•

Integration mode: 21.2×N s/rotation

• Digital radiography mode•Standard mode: 8.3 s/300 mm•

High precision mode: 16.6 s/300 mm•

Ultra high precision mode: 33.2 s/300 mm○ X-Ray Irradiation System*•

MBR-1520R (Hitachi Aloka Medical):•This instrument can be used to irradiate small animals (mice, rats) or cells with
X-rays to study the effects of radiation on organisms, or to breed immunodeficientanimals.

•Tube energy range: 35-150 kV (adjustable in 1 kV steps)•

Tube current: 5/10/15/20 mA•

Beam conditioning filters: 9 choices available

Authorization to use animal experimentation facilities is required to operate the instruments listed
under sections marked by an asterisk (*).