CoolRack - BioCision

BioCision
Consistent.
Reproducible.
Standardized.
September 2011
CoolProducts
CoolProducts enable biological sample consistency,
reproducibility and standardization – user to user, site to
site, trial to trial.
CoolRack®
Error-Free /
Temperature Control
CoolBoxTM
Ice-Free /
Sample Handling
CoolCell®
Alcohol-Free /
Cell Freezing
2
Problem: Lack of Standardization
“The lack of standardized, high-quality biospecimens has
been widely recognized as one of the most significant
roadblocks to the progress of cancer research.”
- NIH / NCI / OBBR
• 
Worldwide effort to develop new standards for collecting,
handling, and processing pre-analytical samples (blood,
tissue, tumor, other biological specimens or sample
materials)
• 
• 
• 
• 
• 
• 
EU: http://www.devicelink.com/ivdt/archive/09/04/003.html
NCI/NIH: http://biospecimens.cancer.gov/practices/default.asp
NIH Office of Biorepositories and Biospecimen Research 2010 Symposium:
http://www.brnsymposium.com/meeting/brnsymposium/2010/
BioCoR: http://biocor.umn.edu/
Duke: http://web.genome.duke.edu/cores/biorepository/protocols/
Molecular Epidemiology Workshop, Shanghai, 2008
3
CoolRack
CoolRack
Error-Free /
Cooling, Freezing or
Thawing
4
Ice Cooling
“BioCision’s proprietary and patented portable bench top tools reduce
variability and ensure consistency in clinical sample handling. How
samples are handled prior to analysis is critical to the outcome of experiments in
basic and clinical research. "Biotech International” (November 2010)
Current method of cooling samples in ice. The
random layout can result in mis-identification,
errors in aliquoting and temperature variability.
CoolRack modules instantly introduce
organization and consistent temperature to all
samples (+/- 0.1oC).
5
Solution: CoolRack
Reproducible and standardized cooling, (snap)freezing, thawing
CoolRack in
water bath
CoolRack
in dry ice
Five consecutive freeze/thaw cycles using a CoolRack module show
highly reproducible freezing and thawing profiles
Biotech International (November 2010)
6
How does it work?
• 
CoolRack modules are made of a novel thermoconductive alloy material
• 
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Thermo-conductivity is the transfer of heat from
higher temperature region to lower temperature
region
• 
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Replaces “direct immersion” into temperature
sources such as ice, dry ice, LN2, water bath
CoolRack on Ice: Heat from the relatively warmer
CoolRack module is transferred to the ice (dry ice,
LN2) until equilibrium is reached
CoolRack in Water Bath: Heat is transferred from
water bath toward relatively cooler CoolRack until
equilibrium is reached
CoolRack novel alloy evenly distributes the
temperature across all wells providing very uniform
and consistent temperature to all samples
7
CoolRack
Versatile
• 
CoolRack rapidly adapts to any temperature
• 
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from -150oC through 100oC+
cooling (ice), snap-freezing (dry ice, LN2),
thawing/incubating (water bath, heat block)
Uniform temperature
• 
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+/-0.1oC temp uniformity to all wells
standardize assay to assay, user to use, site to site
Patent-pending novel alloy construction
• 
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very rapid thermo-conductivity/adaptivity
temperature assurance – no mess, no uncertainty
8
Modular
For a variety of biological specimens
CoolRack
tube holders
CoolSink
Plate holders
CoolTray
platforms
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Versatile
For a variety of applications
-150oC
-78oC
0o to 4oC
37oC +
Liquid Nitrogen
Dry Ice
Ice
Water Bath
•  Eliminate direct immersion
into LN2
•  All tubes are upright
•  Same freezing profile for all
samples
•  ThermalTray provides
elevation of samples &
increased distance between
hands and LN2
•  Rapidly adapts from ambient
(25oC) to LN2 temp (-150oC) in
12-14 minutes
•  Replaces direct contact with
dry ice
•  Eliminates ethanol bath on
the bench – cost savings,
reduced hazard & waste
•  CoolRack PF provides equal
or faster snap-freezing than
ethanol slurry
•  Standardized freezing minimized standard deviation in
freeze profiles
•  Improved work flow
•  Adapts to -78oC in 5-7
minutes
•  Gain organization & stability
in ice
•  Even temperature distribution
to all samples even as ice melt
•  Labels stay dry and legible for
better sample identification
•  Reduced risk of sample
contamination
•  Rapidly goes from ambient to
<4oC in 90 seconds
• Using ThermalTray, 10-15
hours of <4oC with one pan of
ice
•  Keeps tubes out of water –
reduced contamination, labels
stay dry
•  Consistent thawing profile for
all samples
•  All wells at identical
temperature - eliminates edge
effect on multi-well plates
•  Stable support of tubes &
plates
BioCision’s tools are unique and replace very rudimentary methods of sample preservation
(freezing, snap-freezing) and handling (cooling, thawing)
10
CoolRack Performance
Cooling Rate - Ice
§  Ambient to < 4oC in 60-90 seconds
§  4-6 hours of use when placing CoolRack directly on
ice
§  Up to 14 hours cooling with a single pan of ice with
ThermalTray
Cooling rate on ice
Snap-Freezing Rate – Dry Ice
§  Equal or better freeze rate than dry ice/
ethanol bath
§  No alcohol required – cost savings, reduced
hazard
§  Consistent freeze profile for all samples
Freeze rate on dry ice
Standard deviation
11
CoolSink Performance
Ø 
Ø 
Multi-well plates placed directly on ice for
cooling do not achieve critical < 4oC
temperature in any of the wells (Fig. 2, A)
CoolSink modules evenly distribute the ice
temperature to all wells (Fig. 2, B and C)
• 
• 
All wells are <4oC
Curvature of the plate accounts for center
well variability (Fig.1)
Figure 1. Variance in height of undersurface of 96-well flatbottom plate. The x and y axes represent the well number
and letter, and the z axis represents the relative deviation of
the surface in units of 0.001 in. Data was collected using a
ball dial indicator with a resolution of 0.0001 in.
Figure 2. Final equilibrium well temperature for a 96-well flat bottom plate
in direct contact with (A) crushed ice, (B) CoolSink 96Fwith a dry interface,
and (C) CoolSink 96F with aqueous conductive medium in the interface.
Colors represent 0.5°C temperature intervals of the corresponding plate
wells. The white cell represents the well that was fitted with the
thermocouple probe.
Published in Biotechniques, November 2010
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Kits and Accessories
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CoolBox
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CoolSystem Kits
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Ice-free cooling container
Application based
Tube based
Accessories
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Ice pans
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o
Temperature strips (1-8 C)
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Rectangular (1L, 4L, 9L) and round (4L)
LCD display adheres to all thermo-conductive modules
CoolMat histology cutting mat
• 
Protects surface of ThermalTray and scalpel blade
13
Customer Feedback - CoolRack
Steffan Vartanian - Genentech
Don’t you hate it when you spend all morning preparing RNA or harvesting lysates, only to
come back from lunch to find your precious samples drowning in a pool of melted ice? ……
But after using the Thermal Tray and CoolRack a few times, my soggy-sample days are
officially over! Not only do these tools keep your samples chilled for hours, but they also
keep everything organized, safe, and secure. BioCision's products are very well made, well
designed, and just plain look cool. Highly recommended!
Hans B - Perkin Elmer
We have been using the CoolRack for snap freezing our samples for the past weeks with
great success. Since we were on a tight schedule we performed some validation in house
and were very pleased with the results.
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CoolBox
CoolBox
Ice-Free /
Sample Cooling &
Freezing
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Alternative to Ice Cooling
Current method of cooling samples in ice. Directly inserting tubes into
ice can result in temperature variability, lost or illegible labels, misidentification, errors in aliquoting and increased risk of contamination.
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CoolBox eliminates ice
Consistent ice-free cooling and freezing
•  cartridges provide up to 10 hours of ice-free 0.5 – 4oC cooling
•  continuous +/- 0.1oC consistency to all wells
Cool, freeze or snap-freeze
•  five temperature options using cartridges,
dry ice or LN2
•  portable
•  accommodates variety of tubes & plates
Highly recommended for
•  PCR and RNA work, cell culture, cooling in biosafety cabinet, GMP/GLP
suites and vivarium, bio-hazardous containment, short transport
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CoolBox 30 & CoolBox Microplate
CoolBox Microplate System (for plates)
• Ice-free SBS-format plate & tube cooling &
freezing
o
• Includes CoolBox, lid, blue +2 C cartridge
• Accommodates all SBS format modules CoolSinks, CoolRack PCRs, CoolRack 1mlx96
• Sold separately OR with CoolRack/CoolSink
CoolBox 30 System (for tubes)
• Ice-free tube cooling / freezing
o
• Includes CoolBox, lid, blue +2 C cartridge, red
stage, purple insulator pad
• Accommodates all 30-well CoolRacks
• Sold separately OR with CoolRack
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CoolBox Performance
mmmm
-25oC with thermal regulator
pad placed between CoolRack
and dry ice
Snap-freezing at -78oC with
CoolRack directly on 200cc dry
ice
NOTE: Performance may vary depending on ambient
conditions, starting temperature and other specific set-up
criteria. See instructions for complete information.
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CoolBox Advantage
CoolBox
Gel-Filled Container
Well-to-well Temperature Variance:
+/-0.1oC
Well-to-well Temperature Variance:
Up to 4oC
Cooling Media:
LN2, Dry ice, Wet ice, Cartridge
Cooling Media:
Gel
Temperature range:
-150oC through +150oC
Temperature range:
Fixed at -20oC or 0oC
Tube & Plate Formats:
0.5ml conical, 1.5ml conical, 1.5-2.0ml
cylindrical, 12.5mm dia cryos, mass
spec tubes, PCR 96- and 384-well
plates, 6-, 12-, 24-, 48-, 96-well plates
Tube & Plate Formats:
0.5ml conical, 1.5ml conical, PCR tubes
Cooling Duration:
10h (2ºC), 6h (-12ºC), 5h (-25ºC),
6h (-78ºC)
Cooling Duration:
8h (0ºC), 3h (-21ºC)
Down Time:
None – switch cartridge with another
frozen one from the freezer or replenish
dry ice, LN2
Down-Time:
4-6 hours to re-freeze gel-filled tube rack
CoolBox provides temperature uniformity and versatility
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Application-based kits
Protein / DNA CoolSystem – Ice Free
Microbiology CoolSystem
Tissue Dissection CoolSystem
PCR Prep CoolSystem
Protein / DNA CoolSystem
21
Customer Feedback - CoolBox
Fraser Moss Ph.D. - Case Western Reserve University, Dept. of Physiology and
Biophysics, President Scientist Solution
BioCision has now eliminated the ice factor and have the CoolBox in which you can keep the
96 well CoolRack…..I'm using it to keep my RNA cold for oocyte injections right now. I'll be
using it to assemble my PCR reactions in the not to distant future. This has been a great new
little toy to have on the bench recently because our departmental ice machine broke in April
and still is not fixed!
Laura Matthews - Sarah Cannon Research Institute (SCRI)
I am writing regarding the demo CoolRack and CoolBox that we have been testing. They are
both great products. The CoolRacks have eliminated the mess of mushy cardboard freezer
boxes. The samples also freeze much faster in the CoolRacks. The CoolBox was especially
helpful in the transport of fresh biopsies on dry ice from one location to another
22
CoolCell
CoolCell
Alcohol-Free /
Cell Freezing
23
-1oC/min Controlled-Rate Cell Freezing
Highest reproducibility
vs. other methods
• 
• 
• 
-1oC/minute freeze rate in -80oC freezer
Identical freezing profiles every time
Format: 1ml – 5ml cryo vials or injectable vials
Highest cell viability
• 
Cell lines, PBMC, primary cells, stem cells,
yeast, algae
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Requires no isopropanol, no fluids
No on-going expense
No toxic waste
No alcohol-induced variability
Alcohol-free rate control
24
Controlled vs. Uncontrolled Freezing
Controlled
(-1oC/min)
Uncontrolled/
Inconsistent
Post-thaw viability
Controlled freezing
reproducibly minimizes cell
disruption and improves
viability
Allows variation in cell
viability and function with
unpredictable result batch to
batch
Freezing artifacts
Reduces freeze selection of
sub-populations of cells and/
or biomarkers
Selects for sub-population of
cells and/or function that can
best survive the freezing
conditions
Gene/biomarker
expression & function
Ensures cells will be
recovered in a predictable &
consistent state (advantage
when working with precious
and small samples)
Cells will be recovered with
an unpredictable and
inconsistent state
Cell therapy &
diagnostic
Essential to cell therapy and
diagnostics
Roadblock to consistent
diagnostic & clinical outcome
25
Multiple Formats – Same Function
Consistent -1oC/min freezing
1ml and 2ml cryo vials
12 wells
1ml and 2ml cryo vials
30 wells
- Vial module can be easily
transferred to cryo storage
box
4ml and 5ml cryo vials
12 wells
2ml serum vials
12 wells
10ml serum vials
6 wells
26
What is CoolCell?
Passive Controlled Cell Freezing*
• 
• 
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Radially-symmetric thermal exchange to
ensure uniform cooling profiles to all vial
positions
HD Materials used in aerospace industry
for lightweight & insulative properties
Highly Thermo-conductive core
Conduction and/or micro-convection air
control (FTS30)
* Patents Pending
27
CoolCell: Thermo-Dynamic Consistency
Consistent freezing, reproducible results
Five consecutive freeze
cycles in -80oC freezer
using CoolCell (12-well,
2ml cryo vials). Cooling
profiles and phase
change are identical
every time.
28
Advantage CoolCell
CoolCell
Isopropanol Container
No IPA/No maintenance
• No fluids
• No pre-cooling required
• No hazardous waste
100% Isopropanol
• On-going purchase, maintenance, hazardous waste due to
IPA degradation
• Pre-cool hazardous alcohol in refrigerator
No variability
• All vials have uniform freeze rate every time
• Radially symmetric design ensures consistency to
vials and freeze runs
Inconsistent freeze rate
• Each run is slightly different due to IPA degradation
• Two circles of wells – uneven heat removal from vials
No hidden cost
• Once you buy it, you’re done paying for it!!
~$350/year maintenance per unit
• Change IPA every five uses + hazardous waste disposal
• 10 units = $3,500 maintenance cost EVERY YEAR
Lid opens easily
• Cap comes off easily when frozen
• Not cold to the touch, shatter-proof
Screw cap very difficult to remove when frozen
• Protective glove required when handling frozen unit
• Unit is cold, slippery, breakable
Highest post-thaw cell viability & function
Ready to use again in 5 minutes
Takes >1 hour for device to return to room temp
Low impact on freezer
• 1/3 the heat impact on freezer compared to alcoholfilled units
Large thermal mass impacts local freezer area
• Large heat capacity removed from alcohol impacts nearby
samples
29
Cell Line Performance Comparison I
CoolCell vs. “Mr. Frosty”
% GFP transfection efficiency
% Viability
100%
100%
80%
80%
Mr.Frosty
60%
CoolCell
(12 samples)
40%
CoolCell
(30 samples)
20%
Mr.Frosty
60%
CoolCell
(12 samples)
40%
CoolCell
(30 samples)
20%
0%
0%
HeLa
CHO-K
K562
NIH3T3
HeLa
CHO-K
K562
NIH3T3
Independent Performance Analysis by LONZA/Germany
• 
Identical transfection efficiencies and viabilities after freezing & thawing of cells
• 
CoolCell is more durable than Mr. Frosty (i.e. CoolCell is shatter-proof)
30
Cell Line Performance Comparison II
CoolCell vs. “Mr. Frosty”
Growth performance after 24 hours post-thaw
6
5
4
Mr Frosty
ATP RLU
3
(ViaLight)
CoolCell
(12 samples)
CoolCell
(30 samples)
2
1
0
HeLa
CHO-K1
K562
NIH3T3
Independent Performance Analysis by LONZA/Germany
• 
Identical growth of cells observed 24 hours after thawing
• 
Freezing and thawing is easier and more convenient with CoolCell (no isopropanol)
31
Primary Cells Performance Comparison
CoolCell vs. “Mr. Frosty”
PBMC
HUVEC
92%
CoolCell
Alcohol-based
0
89%
20
40
60
80
100
!"#$%&'(")(''")*+,-"
Figure 1. Post freeze viability, PBMC. 2x107 PBMC cells total were isolated
and split into two vials, at 1x107 cells per vial, in Human Sera with 10% DMSO.
One vial was frozen overnight using a BioCision CoolCell and one in a
commercial IPA-based freezing system (“Mr. Frosty”). Following long term
storage in LN2, cells were thawed and viability counts for each vial were
obtained by the trypan blue stain method.
94%
CoolCell
Alcohol-based
0
82%
20
40
60
80
100
!"#$%&'(")(''")*+,-"
Figure 2. Post freeze viable cell count, HUVEC (n=5). Cells were
resuspended in freezing medium at a concentration of 2x106 cells per ml.
1ml aliquots were proportioned into 1.8ml cryo vials and frozen at -1oC/
minute in either a BioCision CoolCell or in an alcohol-filled freezing unit. Five
vials frozen by either method were rapidly thawed and resuspended in
growth media. Live cell counts were obtained by the trypan blue exclusion
method.
Independent Performance Analysis
• 
In primary human PBMC and HUVEC, freezing with CoolCell produces >90% viable cell
count post-thaw without the use of isopropanol (a flammable, hazardous solvent) which is
used in the alcohol-based method
32
Stem Cells
CoolCell vs. other common methods
Common un-standardized methods of stem cell cryopreservation
• 
• 
Styrofoam box
Paper towel insulation
Performance comparison with CoolCell
• 
• 
Independent evaluation of CoolCell cyropreservation system initiated
by Roslin Cellab, a leading stem cell research institute
Determine effect of different freeze/thaw methods on critical stem cell
growth parameters (viability)
33
Dead Cells after Freezing
CoolCell vs. other common methods
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5-1:-;/$+2$<-3<$
:-,,9$3=-1$
/>38?;@$$
%!$
%&"'$
%)$
Figure 1. Determination of
amount of dead RC10 cells
(human embryonic stem cell)
u s i n g Tr y p a n B l u e a f t e r
thawing. Comparison of 3
difference freezing methods
(n=3)
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*++,*-,,$
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536-1$7+8-,9$
Independent Performance Analysis by Roslin Cellab
• 
Cryopreservation with CoolCell results in less cell death upon thaw (P<0.05)
• 
CoolCell ensures less variability when freezing multiple vials.
34
Stem Cell Performance Comparison
CoolCell vs. other common methods morphology data
CoolCell
Styrofoam Box
!
Paper Towel
!
!
Figure 2. Human embryonic stem cell line RC10 was observed at day 1 post resuscitation under
x10magnification using three different freezing methods.
Independent Performance Review – Roslin Cellab
• 
Stem cell morphology was visually improved when using the CoolCell compared to other methods.
This complements the data showing the higher viability and proliferative capacity results, as seen
with CoolCell.
35
Stem Cell Proliferation
CoolCell vs. other common methods
'"
Figure 3. RC10 cells count
using Trypan Blue at 3 days
post-thaw using three different
freezing methods (n=3)
&"
!$##%!"34'%
5.6789%
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#"
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/+0$)%1"2$#%
Independent Performance Analysis by Roslin Cellab
• 
Significant increase in viable cell count using a CoolCell (P<0.001)
1 Million cells (33%) more than other methods
• 
• 
Direct impact in various experimental results (i.e. detection of proteins secreted in low levels)
Decrease in patient sample size required for diagnostics, treatment or research
36
Stem Cell Proliferation over Time
CoolCell vs. other common methods
(#$"
Figure 4. RC10 cells count
using Trypan Blue at days 0
and 3 post-thaw using three
different freezing methods.
Average of 3 samples.
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'#$"
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01%!23"
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Independent Performance Analysis by Roslin Cellab
• 
Stem cells cryopreserved with a standardized and temperature controlled CoolCell recover better
and their proliferation rate is much higher
37
®
CoolSystem
CRYO
Kit for ice-free vial preparation and alcohol-free cell freezing
CoolSystem CRYO (Item #BCS-142)
Includes:
CoolBox CFT30 for ice-free vial cooling during preparation in the biosafety cabinet
CoolCell 12-place 2ml vial controlled-rate freezing container
38
™
TruCool
Cryogenic Tubes
Leak-proof caps, individually barcoded
•  Thermally-fused gasket prevents leakage and O-ring contamination
•  Sterile, medical-grade polypropylene – will not discolor upon autoclaving
•  Individually barcoded – no external labeling required (Barcodes are Code 128
and 10 characters)
•  Internal- and external-threading options
39
Customer Testimonials
John Gardner – Roslin Cellab
“This is by far and away the best benchtop technology to enter the field of cell
cryopreservation for decades.”
Rohit G. - Stanford University
“We run a registry, in which large amounts of PBMCs are processed for long-term
cyropreservation.………Overall, the CoolCell has proven to us to set a new bar in
cryopreservation.”
Sam Knight - Ceramisphere (Australia)
“I spent nearly a year getting terrible viability-recovery on cell freezing with dry ice (using an
alcohol device) followed by storage in LN2, and this on HeLa cells which aren't exactly hard to
store. I then bought a CoolCell and now I can just use standard cryo-preservation techniques
(DMSO, 20% NBS) and get nearly 100% viability on thawing.”
40
CoolCell Summary
§ 
Controlled cryopreservation results in
§ 
§ 
greater cell recovery (less dead cells)
better “functional” recovery (higher rate of proliferation)
Highest reproducibility between samples
§  Less selection of cell subsets
§  Smaller sample size (patient sample/biopsy, hard to
transfect cells, flow cytometry sorted cells, generation
of primary cells)
§  Easy to use, no toxic waste, easy transport to freezer
farms
§ 
41
Summary BioCision
Pre-Analytical Sample Standardization
Sample handling has become increasingly important
• 
• 
Growing number of multi-national companies, labs, studies
Increasing use of biomarkers and highly sensitive analytical methods
BioCision = Consistency. Reproducibility. Standardization.
• 
• 
• 
• 
Test to test
Researcher to researcher
Lab to lab
Site to site
Watch the VIDEO
42
Genentech Amgen Pfizer
Roche
Harvard
MIT
CalTech UCSF Glaxo Bristol Myers Squibb Novartis
Translational Genomics
Gilead Sciences
Merck
MedImmune ATCC Genpharm
Elan Pharmaceuticals
Centocor Boehringer Ingelheim Oncomed NIH/NCI
Takeda Pharmaceuticals
CDC
Covance
XOMA
Quest Diagnostics Otsuka America Pharmaceuticals
Salk Institute Burnham Institute Scripps Institute
Stanford University Yale Genzyme
EMD Serono
Shire HGT
Alnylam
Wyeth
Cell-Systems
Jackson Labs Beth Israel Deaconess Hospital HHMI
Whitehead Institute
Harvard Systems Biology
Exelixis Charles River Labs Sanofi Pasteur Penn State
Lawrence Berkeley Lab Batelle
UC Berkeley
Cedars Sinai Hospital Cleveland Clinic IDEXX Biogen
Perkin Elmer Genetics Blood Systems Research Inst.
Thank you
Copyright 2012. Patents pending. All rights reserved.