Sunteți pe pagina 1din 4

Data

Data Sheet
Sheet

Guava® ViaCount® Assay


Rapid, accurate cell counting and viability assessments
The Guava ViaCount Assay is fast becoming the new
standard for viability and cell counting. In this simple
no-wash, mix-and-read-assay, you can accurately
obtain absolute total cell counts, perform viability
assessments, and determine apoptotic percentages—
all from tiny samples.
You’ll enjoy several advantages over traditional
methods, including greater accuracy, reproducibility,
and speed. This assay eliminates the user-to-user
variability characteristic of trypan blue hemocytometer
counting. As a result, you have significant increases in
counting reproducibility. In addition, the Guava ViaCount
Assay is more accurate than hemocytometer counting
or impedance-based counting over a wide range of
cell concentrations. Cell counting with this assay has
also proven to be up to 10 times faster than manual
hemocytometer methods. Unlike traditional cytometer
counting, you don’t need to use expensive reference
beads because of Guava’s unique capillary technology.
Features Cell counting and viability assessments can be used
Assay: in a variety of applications, such as cytotoxicity studies,
• Simple no-wash, mix-and-read procedure PBMC counting, rapid apoptosis assessment, and
• Analyzes viability of individual cells increasing reproducibility in cell-based assays.
• Delivers direct, absolute cell counts and population
percentage
• Counts up to 10 times faster than manual methods
Debris Viable Cells Apoptotic Dead Cells
• More reproducible than traditional tests
Forward Low High High High
Samples: Scatter
• Uses small samples in tubes or a 96-well plate Nuclear Neg High High High
• Handles low-density and small-volume cell samples Stain

• Works with adherent or suspended cells and mammalian Viability Neg Neg Med High
Stain
and insect cells

Compatibility:
• Runs on Guava EasyCyte™ Mini, EasyCyte Plus, PCA,
and PCA-96 Systems
QUICKLY IDENTIFY APOPTOTIC CELLS
8
With the Guava ViaCount assay, you can easily identify
Observed Count (x106 cells/mL)

Guava PCA ( )
apoptotic cells using the viability dye (figure on front
Hemacytometer ( )
6 Impedance Counter ( ) and figure 2). You can also use an optional apoptosis
gate in the analysis software that counts apoptotic
4 cells separately. This optional gate feature enables you
to use the assay for cytotoxicity, apoptosis induction,
2 or proliferation studies. In these cases, more than a
mere cell number is essential to understanding an
0 experiment’s outcome.
0 2 4 6
By contrast, published studies show that trypan
Expected Count (x106 cells/mL)
blue exclusion assays often overestimate cell counts2,3.
Guava PCA Hemacytometer Impedance Counter This may occur because trypan blue cannot identify
y=0.94x + 0.25 y=0.95x + 0.23 y=1.27x + 0.17
R2=0.966 R2=0.877 R2=0.964
apoptotic cells, and so they are added to the count of
viable cells.

Figure 1. Guava ViaCount assay provides accurate cell counts.


Comparisons with an impedance counter and manual trypan blue
IMPROVE YOUR CELL-BASED ASSAYS
hemocytometer counting indicated greater linearity and accuracy over a Accurate cell counts and viability measurements make
range of cell concentrations. Expected cell counts were determined using other functional cellular assays more reproducible
an FDA-cleared reference counting bead method 1.
because they provide a more realistic indication of how
many healthy cells are present. For example, ELISPOT
assays show a marked reduction in CVs when cell
HOW THE GUAVA VIACOUNT concentrations are measured with the Guava ViaCount
ASSAY WORKS assay compared to those done with trypan blue
The ViaCount assay uses a proprietary mixture of two hemocytometer measurements (Figure 3).
DNA binding dyes to provide sensitive, accurate
detection of viable, apoptotic, and dead cells. COMPATIBLE WITH MANY CELL TYPES
A membrane-permeant dye stains all nucleated The Guava ViaCount assay accurately determines
cells, leaving cellular debris unstained so that you can viability and counts for a large variety of cell types.
focus on whole cells. A membrane-impermeant dye Tested on a broad selection of mammalian cell lines
stains only damaged cells (i.e. those with breaches in (including human, mouse, rat, hamster, and monkey). As
the plasma membrane), thus indicating apoptotic and a plus, the assay also works with both primary cells and
dying cells. whole blood.

2000

1800
Manual via Trypan Blue
Guava ViaCount
Live 1600
Nucleated Cells

Dead
SFC/106 cells
Viability Stain

1400
Dead

Live 1200 Mean


Apoptotic 1 stdv
2 stdv
Apoptotic 1000 3 stdv

800
Viability Stain Annexin V
600
1 2 3 4 5 6 7 8
Assay Number

Figure 2. The blue population of cells show a significant amount of


annexin V staining (bottom plot), indicating that intermediate levels of Figure 3. Increased reproducibility in ELISPOT assays when using
staining with the viability dye correlates with apoptosis. Guava ViaCount assay for cell counting.

2
EASY-TO-USE SOFTWARE ADDITIONAL REAGENTS
Guava’s ViaCount Software Module makes data FOR GREATER FLEXIBILITY
collection and analysis even easier. It runs the entire Created to address more difficult cell counting
assay without any need for user input, eliminating samples, Guava ViaCount Flex reagent is a more
subjectivity. Sample analysis is completed and the data concentrated formulation of the ViaCount reagent. It
are automatically exported to a time-stamped works well for low density cell cultures since it can be
spreadsheet in seconds. Plus, Guava’s new EasyFit added in small volumes to avoid further sample dilution.
Software is included for computer-aided population This feature also makes ViaCount Flex reagent ideal for
analysis. cell counting and viability assessment in 96-well plates.
The module displays data in user-friendly formats For clumped samples, use the ViacCount Cell
so you can monitor your experiment or read data right Dispersal Reagent, which uses enzymes to gently
off the screen. By measuring the staining intensities of disaggretate cells. This helps to improve the accuracy
the two dyes, our software represents each cell and precision of cell counts.
counted as a dot on the Cell Dotplot. Cell counts and
percentages are calculated and displayed real-time on ViaCount Flex Reagent
the screen. The optional apoptosis gate (purple line) • Low density cell samples that are too diluted to be
can easily be unchecked if only the viable population is counted with standard hemacytometers
relevant to your research. • Cell lines that stain heterogeneously (facilitates gate
settings by providing tighter staining patterns)
Benefits:
• Easier to learn and use Viacount Flex Reagent for Challenging Samples
• Get standardized results even with multiple users • Non-mammalian cell lines (such as SF9 insect cells)
• Spend less time optimizing instrument settings and other cell lines with unusual culture conditions
• Improve your productivity
Guava ViaCount Cell Dispersal Reagent (CDR)
• To reduce clumping in adherent cells

A. Easy viewing of
real-time statistics
for cell counts and
concentrations of
original sample.

B. Optional apoptosis A
gate (purple line)
identifies apoptotic
cells (intermediate level
staining, red circle).

C. Real-time counts of
cells in all populations
(apoptotic, dead, viable) 2B
1B
with calculated cell
C
concentrations.

3
ORDERING INFORMATION
Description Catalogue No.

Kits & Reagents


Guava ViaCount Reagent Kit (100 tests) 4000-0040
Guava ViaCount Reagent Kit (600 tests) 4000-0041
Guava ViaCount Flex Reagent for Challenging Samples (100 tests) 4500-0110
Guava ViaCount Cell Dispersal Reagent (100 tests) 4700-0050
Guava ViaCount Flex Reagent (500 tests) 4700-0060

Software
ViaCount Software, PCA System 0500-1030
ViaCount Software, PCA-96 System 0500-0580
ViaCount Software, EasyCyte MiniSystem 0500-1470
ViaCount Software, EasyCyte Plus System 0500-2000

References To learn more about our integrated solutions for


1. Data provided courtesy of Amy Hirst, flow cytometry, please contact Millipore or Guava
Raven Biotechnologies, Inc.
Technologies. The chart (below) shows which company
2. Mascotti, McCullogh and Burger; Transfusion
serves your organization.
40:693-696, 2000.
3. A
 ltman, Rander and Rao; Biotechnology Progress
9:671-674, 1993. To stay up-to-date on our joint product and
4. O
 gasawara K, et al. Impairment of NK cell function by NKG2D service offerings, please visit
modulation in NOD mice. Immunity. 18(1): 41-51 (2003). www.millipore.com/flowcytometry.
5. A
 rase H, et al. Direct recognition of cytomegalovirus by
activating and inhibitory NK cell receptors. Science.
296(5571):1323-1326 (2002). Guava Flow
Assays Cytometry
6. C
 erwenka A, et al. Cutting edge: the minor histocompatibility Audience & Kits Systems Service
antigen H60 peptide interacts with both H-2Kb and NKG2D.
Journal of Immunology. 168(7):3131-3134 (2002). Academic &
7. O
 gasawara K, et al. Inducible costimulator costimulates Europe Government n n n
cytotoxic activity and IFN-gamma production in activated & North
murine NK cells. Journal of Immunology. 169(7):3676-3685 America Pharma &
(2002). Biotech l l n
8. P
 hi-Wilson J, et al. A technology for the rapid acquisition of
cell number and viability. American Biotechnology Laboratory. All Other Countries n n n
pp 34-36 (2001).

n Millipore Corporation l Guava Technologies

www.millipore.com www.guavatechnologies.com

Millipore is a registered trademark of Millipore Corporation. The M mark is a trademark of Millipore Corporation. Guava, the Guava logo, Guava ViaCount,
and Guava Technologies are registered trademarks of Guava Technologies, Inc. EasyCyte is a trademark of Guava Technologies, Inc.
Lit. No. DS0125EN00 07/08 Printed in U.S.A. BS-GEN-08-00631 © 2008 Millipore Corporation, Billerica, MA 01821 U.S.A. All rights reserved.