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T H E P O W E R O F E A S Y & A F F O R D A B L E
F L O W C Y T O M E T R Y & C E L L C O U N T I N G
The desire to gain a systems perspective by analyzing whole cells has fueled a rising interest in cell- and bead-based applications for basic, applied, and clinical life science research, including drug discovery and development efforts. By studying intact cells, researchers can gain critical insights into the interactions among biological pathways. Further, they can leverage cell studies to expedite target validation, lead optimization, process development, toxicity determinations, clinical trials, and much more.
Accelerate discovery and increase productivity. Guava® Assays can be applied to a wide and ever-expanding range of applications.
T H E P O W E R O F E A S Y & A F F O R D A B L E
F L O W C Y T O M E T R Y
& C E L L C O U N T I N G
2
Many assays. One platform.
Direct, absolute cell counts and viability
Apoptosis• Annexin V • TUNEL • Pan caspase • Caspases 3/7 and 8 • Mitochondrial membrane potential
GFP expression
Cell cycle
Antigen detection
Cell toxicity
Cell proliferation
Cell painting
Cell signaling
Antibody quantification
Multiple cell types.• Transformed • Mammalian• Primary • Insect• Suspension • Yeast • Adherent
Beads.
Cell counts and viability• Cytotoxicity screening• Cell culture optimization• Bioprocess monitoring
Apoptosis• Cell line characterization• Compound screening• Bioprocess monitoring
Antigen detection• Compound profiling• Protein/antigen expression analysis• Hybridoma screening• CD marker detection• Cytokine quantification
Cell cycle• Cell line characterization• Compound screening• Bioprocess monitoring
Cell tracking• In vitro and in vivo partitioning• Cell toxicity• Cell proliferation
Fluorescent proteins• GFP screening• Combined transfection efficiency and viability
Bead-based assays• Antibody quantification
“[Guava® Systems] have great value not only for assay development and drug lead optimization, but also as credible screening instruments giving high content readouts from a single well.”
—JillGregory,StaffScientist,GenzymeCorporation Drug Discovery
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Flow cytometry on demand.The Guava® PCA and PCA-96 Systems as well as the Guava EasyCyte™ Mini and Plus Systems have been designed to meet the needs of today’s life scientist. These on-demand systems, together with an ever-growing suite of assays, deliver powerful solutions for counting, profiling, and screening a most valuable commodity—your samples. Whether you focus on scientific applications, routine screening, cell counting, or commercial production, Guava Technologies has the solution that is right for you.
Turnkey solutions.Guava Technologies has streamlined and simpli-fied fluorescent cell- and bead-based assays by integrating instrument, software, and reagents into targeted, turnkey solutions. Samples in. Answers out. Next study. The assay develop-ment has been done for you, so you can concentrate on getting results.
O P T I O N A L : - S I D E S C AT T E r - 4 T H CO LO r D E T E C TO r
Guava EasyCyte™ Plus System
I N T U I T I V ES O F T WA r EI N T E r FAC E
M I N I M A LWA S T E
Guava® PCA System
G r E E N L A S E r
M I C r O P L AT E O r T U B E S
Guava® PCA-96 System
Guava EasyCyte™ Mini System
B LU E L A S E r
Do more.Each Guava® Assay meets exacting standards of robustness, ease-of-use, and affordability, while requiring only a few microliters of sample. And, Guava Systems are designed to be very easy to use, so you can be up and running within a few hours! Sophisticated cell analysis has never been easier.
Do it right.Guava Systems combine fluorescence-based techniques and high quality optical elements to maximize assay sensitivity. And, automated software routines reduce operator-induced variability. The result? Accuracy and reproduc-ibility.
Do it fast.With an integrated system of instrument, software, and reagents from Guava Technologies, you don’t have to rely on core labs to run your sophisticated cell analysis assays any longer. You can conduct your own studies in your own lab on your own schedule.
PCA PCA-96 EasyCyte Mini EasyCyte Plus
Laser Green Green Blue Blue
Tiny footprint
SCATTER Forward (FSC) Side (SSC)
—
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optional
optional
Tube sampling
Automated 96-well microplate sampling — —
Small sample size (<20 µL)
Minimal waste (<50 mL per day)
GreenFLUORESCENCE Yellow
DETECTORS Red Near infrared
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——
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optional
Four systems meet a variety of needs.
O P T I O N A L : - S I D E S C AT T E r
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Trypan Blue
Guava ViaCountAssay
Trypan Blue
Guava ViaCountAssay
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Guava ViaCount
Trypan Blue/ Hemacytometer
T H E P O W E R O F C H O I C E .M A N Y A S S A Y S . O N E P L A T F O R M .
Guava Technologies has streamlined and simplified cell- and bead-based assays by integrating instruments and software with optimized reagents. Now, you can conduct cell analysis studies from your bench top, on your schedule. Now, you have the ability to generate data wherever and whenever you want to make better, faster decisions and maximize your laboratory’s productivity.
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Cell cycle.Determining the distribution of cells across the phases of cellular replication and division is done easily with the Guava® Cell Cycle Assay. In addition to providing G0/G1, S, and G2/M phase percentages, this assay exports data in industry standard formats to allow more industry standard formats to allow more detailed analysis.
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Trypan Blue
Guava ViaCountAssay
Trypan Blue
Guava ViaCountAssay
Low
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95%
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Reproducibility
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Guava ViaCount
Trypan Blue/ Hemacytometer
As shown above, the Guava ViaCount Assay provides superior reproducibility over manual cell counting with Trypan Blue in cell cultures with low viabilities or low cell concentrations.
0 5 10 15 20 25 30
Trypan Blue
Guava ViaCountAssay
Trypan Blue
Guava ViaCountAssay
Low
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100%
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Accuracy
Reproducibility
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Guava ViaCount
Trypan Blue/ Hemacytometer
At lower viability levels, the Guava ViaCount Assay provides more accurate assessments of cell culture viability than Trypan Blue dye exclusion assays using a hemacytometer.
Cell counting and viability.Speed, ease, and depth: Unlike other viability assays, the Guava ViaCount® Assay not only Guava ViaCount® Assay not only Guava ViaCount® Assaydetects viable and dead cells, but also simul-taneously assesses the apoptotic cell fraction. requiring only a single reagent, it is a simple no-wash, mix-and-read assay that rivals the fastest of similar assays. Moreover, its versatility extends its applicability across a wide range of cell types, including suspended and adherent, mammalian and non-mammalian, and many others.
Multicolor antigen detection.Guava Express® Cell Surface Antibodiess® Cell Surface Antibodiess are fluorophore-conjugated antibodies that have been optimized for use on most Guava®Systems. The antibodies provide quick, easy staining of human CD markers and allow for rapid multicolor immunophenotyping of hematopoetic cells. The Guava Express® Assayand Guava Express® Plus and Pro Software Modules allow you to use your own primary conjugates or our second-step reagents to detect surface and internal receptors or antigens on single cells. This is ideal for hybridoma-screening, immunophenotyping, and other 2-, 3-, or 4-color assays.
The Guava EasyCyte Plus System offers a convenient platform for exploring apoptosis biology, while addressing important applications such as endpoint analysis and mechanism of action studies.
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Dual staining of lysed whole blood with anti-CD45rA-PE and anti-CD3-PECy5 identifies naïve T cells (CD3+CD45rA+), memory T cells (CD3+CD45rA-), and B cells (CD3-CD45rA+). Cells in the lymphocyte gate were used for this analysis.
Based on the detection of fluorescent DNA-binding dyes, the ViaCount Assay offers far greater accuracy and reproducibility than traditional Trypan Blue-based methods, while using a much smaller cell sample. Because it can identify both viable and apoptotic cells, the ViaCount Assay can be used for cytotoxicity screens, proliferation assays, and cell culture monitoring. In addition, ViaCount reagents can assess cell health in any cell-based assay.
10e4
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CD3-PECy5 (RED-HLog)
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Culturecells
Induce
Split cells & selectdesiredassays
Guava® Cell Cycle Assay
Guava® Caspase 3/7 Apoptosis Assay
Guava® Caspase 8 Apoptosis Assay
Guava® Multi-Caspase Apoptosis Assay
Guava Nexin® Annexin V Apoptosis Assay
Guava® Mitochondrial Depolarization Apoptosis Assay
Guava® TUNEL Apoptosis Assay
Endpoint analysis
Mechanism of action
“The Guava® PCA is a major advance in our ability to rapidly and easily measure most of the parameters important in cell biology.” —JennieMather,CEOandPresident,RavenBiotechnologies,Inc.
Cancer Therapeutics
Induction of Early to Mid-Apoptotic Cells
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Mebendazole
Albendazole
Helenine
Compound dose response curve on Jurkat cells. Jurkat cells were treated with the indicated compounds at a range of concentrations for 24 hours before testing for apoptosis induction using the Guava Nexin Assay.
Mitochondrial membrane potential. Designed to save time and effort, the Guava® Mitochondrial Depolarization Assay is a no-wash, mix-and-read assay to monitor changes in mitochondrial membrane potential (∆ψ), the loss of which is often associated with early apoptosis. Collapse of this potential may coincide with the opening of the mitochondrial permeability transition pores, leading to the release of cytochrome C into the cytosol which triggers downstream events in the apoptotic cascade. Addition of the dead cell dye, 7-AAD, allows you to easily discern healthy, polarized cells from the apoptotic/depolarized and dead cells.
Apoptosis.
In a process as complex as apoptosis, multiple methods of assessment are essential to identify
cell populations in various stages of cell death. The Guava® Apoptosis Assay Suite contains several complementary assays that can all be done on a single platform, quickly and easily.
Annexin V. Designed to save time and effort, the Guava Nexin® Assay is a no-wash, mix-and-read assay to identify cells in early- to mid-stage apoptosis. It is one of the fastest and easiest apoptosis assays available. Using Annexin V conjugated to a fluorophore, this assay detects the transfer of phosphatidyl serine from the internal to the external face of the plasma membrane during the initial stages of apoptosis. Addition of the dead cell dye, 7-AAD, allows you to easily discern early apoptotic cells from late apoptotic /dead cells.
Caspases. By virtue of fluorochrome-conjugated caspase inhibitors in combination with a viability dye, Guava® Caspase Assays detect the activation of various caspases during the apoptotic process. Up to four separate populations (viable, early-to-mid apoptotic, late apoptotic, and dead) can be resolved in a single assay. These assays detect simultaneous activation of multiple caspases, caspase 3/7, or caspase 8.
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The Guava Mitochondrial Depolarization Assay can distinguish depolarized Jurkat cells, which fluoresce mostly green, from healthy uninduced cells, which fluoresce mostly orange (overlay).
Fluorescent proteins.
Screening clones of Green Fluorescent Protein (GFP) transfectants is now quick and easy: Guava Express® Plus or Pro Assays automate single cell GFP detection in 96-well plates and combine it with viability assessment, apoptosis, or detection of a second (Plus) or third (Pro) marker. The Guava Express Plus or Pro Assays eliminate the need to test each clone individually since an entire plate of transfectants can be analyzed in a single experiment.
Guava Express Plus & Pro Assays make screening for Green Fluorescent Protein (GFP) and other fluorescent proteins fast and easy.
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The Guava TUNEL Assay easily distinguishes and quantifies the apoptotic cell population, as seen in this sample of HL60 cells treated with 20 µM camptothecin for 4 hours.
Up to four cell populations (viable, early-to-mid apoptotic, late apoptotic, and dead) can be detected using tilting markers in the Guava® Multi-Caspase Assay.
TUNEL. This classic assay for apoptosis-induced DNA fragmentation has been reworked to make it more convenient and accessible. The Guava® TUNEL Assay detects late-stage apoptotic cells by using sensitive fluorescent probes to detect DNA strand breaks.
“The beauty of the Guava® PCA System is that it generates reliable and reproducible quantification of apoptotic cells.” —MaureenE.Murphy,Ph.D.,DepartmentofPharmacology,FoxChaseCancerCenter
Cancer Research
Cell tracking.Improve your cell tracking with three easy-to-use, non-radioactive assays that are superior to population-based methods. Cell tracking assays from Guava Technologies use a well-character-ized cell tracking dye, CFSE, that diffuses freely into cells and is retained within the cell without affecting cellular function.
Cytotoxicity. When CFSE is used in combination with the dead cell dye, 7-AAD, all of the commonly performed cytotoxicity effector assays (NK, CMC, ADCC) can be performed with the Guava® Cell Toxicity Assay.
Cell proliferation. The Guava® Cell Growth Assay is used to monitor cell proliferation of Assay is used to monitor cell proliferation of Assayprimary cultures out to five generations as well as to discern which portion of the cells are dead when stained with propidium iodide.
Enriched T cells stimlulated with 5 µg/mL PHA, assayed on day 4 and analyzed using the ModFit™ software package from Verity Software House.
Guava Cell Growth Assay
6
Guava Express® Pro Software.
Hybridoma screening.Using quantitative bead-based measurements, Guava RapidQuant™ IgG Assays quantify the levels of soluble IgG in hybridoma supernatants with much less work than traditional assays, such as ELISAs. Designed to be easy and effortless, rapidQuant IgG Assays feature no-wash protocols, superb day-to-day reproduc-
ibility, shorter time to answer, and improved accuracy over other available assays. Both mouse and human rapidQuant IgG Assays are universal—they quantify all IgG subtypes of that species. Further, rapidQuant Assays can
be combined with other assays from Guava Technologies to accelerate hybridoma screening and monoclonal antibody development by allowing all stages of testing to be performed from less than 50 µL of supernatant.
Optimized reagents.Optimized for use on all Guava® Systems, reagents from Guava Technologies are formu-lated to ensure excellent sensitivity and ease-of-use. These unique reagentscan also improve and accelerate your assays by eliminat-ing many of the common preparation stepsrequired with traditional proto-cols. In some cases, samples can be read immediately after stain-ing, with no loss of sensitivity.
Dilutions of various antibodies tested with the Guava rapidQuant Human IgG Assay show excellent correlation with predicted values from ELISAs.
Accurate Prediction of Human IgG Concentrations
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Cytotoxicity data are automatically analyzed and displayed in an easy-to-read format.
Guava Cell Toxicity Assay
Dedicated software.The friendly and intuitive Guava CytoSoft™ Operating Environment and Guava® Assay-Specific Software Modules guide you through setup, maintenance, cleaning, and shutdown procedures as well as data acquisi-tion and analysis. All modules are designed to streamline the experimental process from hypothesis to conclusion. results can be exported to spreadsheets or as industry-stan-dard FCS 2.0 or 3.0 files for further analysis by third-party software. And, all software modules support compliance with 21 CFR Part 11 regu-lations. For multicolor detection, the Guava Express® Pro Software Module allows you to view and analyze up to eight user-selectable histogram or dot plots simultaneously. You can also customize axis labels and statistics, and select multiple marker and gate types. Additional features include histogram overlays, backgating, zoom-to-plot for enlarged views, and batch-file printing.
Cell painting. The Guava® Cell Paint Assay utilizes the cell tracking dye as a single color assay to track target cells within a mixed population for in vivo and in vitro applications.A second, user-defined dye can be added to perform a dual color assay for monitoring protein expression or antigen specificity.
At the heart of Guava Systems is a patented, microcapillary, laser-based technology capable of detecting mammalian and microbial cells and beads. A sample of fluorescently labeled cells is aspirated into a uniquely proportioned microcapillary flow cell. A green or blue diode laser excites the cells—each cell emits signals that are individually detected by photomultipliers and a photo diode. Guava Software Modules show all relevant data and results immediately.
T H E P O W E R O F S M A L L .T H E G U A V A F O R M U L A .
Turnkey solutions for cell analysis from Guava Technologies are based on a superior interplay among reagents, software, and instrument. This sophistication elevates cell analysis and counting to an art—while bringing it down to earth. The result: Tools that are truly a pleasure to use.
The flow cell is easily removed for routine maintenance.
G r E E N L A S E r E x C I T E S
F LU O r E S C E N T LY
L A B E L E D C E L L S
F O r WA r D S C AT T E r
( F S C ) P H OTO D I O D E
D I F F E r E N T I AT E S
C E L L S B Y S I z E
P H OTO M U LT I P L I E r S ( P MT )
D E T E C T F LU O r E S E C E N T LY
L A B E L E D C E L L S
Guava® PCA System
B LU E L A S E r E x C I T E S
F LU O r E S C E N T LY
L A B E L E D C E L L S
SIDE-SCATTEr (SSC)
P H OTO M U LT I P L I E r ( P MT )
D I F F E r E N T I AT E S C E L L S
B Y G r A N U L A r I T Y
Guava EasyCyte™ Plus System
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The technology behind it all.So how does so much power and functionality get packed into such a small space? Credit a unique design that optimizes every available bit of space and eliminates wasted corners. Then, credit a unique microcapillary flow cell technology that eliminates the requirement for complicated sheath flow fluidics. The end result? An extremely compact, easy to use, and easy to maintain system, supporting on-demand cell analysis. Sheathless fluidics translates into fewer cells (<20 µL), less reagent, and
minimal waste (<50 mL per 8 hours), saving you both time and money. Also, since the flow cell is self-aligning and user-replaceable, you can remove it at any time for cleaning and maintenance yourself—no more expense or downtime for service visits. And, of course, the tiny footprint fits into the tightest spots, reserving valuable laboratory bench space for other equipment and uses.
ForResearchUseOnly.Notforuseindiagnosticprocedures.ModFitisatrademarkofVeritySoftwareHouse.Guava,GuavaTechnologies,theGuavaLogoandallothertrademarksarethepropertyofGuavaTechnologies,Inc.©2007GuavaTechnologies,Inc.Allrightsreserved.
Global Headquarters
Guava Technologies, Inc. 25801IndustrialBlvd.Hayward,CA94545USATel:15105761400Fax:15105761500www.guavatechnologies.com[email protected]
Europe & Africa
Guava Technologies, Inc. 3SouthviewBusinessCentreTinwellRoadStamford,LincsPE92JLUKTel:+44(0)1780764390Fax:+44(0)1780[email protected]
Asia-Pacific
Guava Technologies Inc.1502,DLFGalleria,DLFPhase4Gurgaon-122002Haryana,IndiaTel:+911244148247Fax:+91124[email protected]
Expert support and service. All products from Guava Technologies are designed to increase productivity, maximize uptime, and minimize maintenance. But, in the rare instance when you do need assistance, our customer solutions team
is always available to answer questions about our software, instru-ments, reagents, and applications. Individual members of the team specialize in differ-ent disciplines of life
science research to ensure your questions are answered quickly, thoroughly, and completely the first time you call.
Get started today.Discover how products and services from Guava Technologies can accelerate discovery and increase the productivity of your organization. For more information, contact us directly or visit: www.guavatechnologies.com
Specifications PCA PCA-96 EasyCyte Mini EasyCyte Plus
Excitation 532 nm 532 nm 488 nm 488 nm
Emission 580/20 nm 675/20 nm
580/20 nm 675/20 nm
525/30 nm583/26 nm680/30 nm
525/30 nm583/26 nm680/30 nm785/70 nm (optional)
Light scatter Forward Forward ForwardSide (optional)
ForwardSide (optional)
Dimensions 8.5 in (21.6 cm) h 12.6 in (32 cm) w 14.3 in (36.3 cm) d
13 in (33 cm) h 16 in (40.6 cm) w 22.5 in (57.2 cm) d
9 in (23 cm) h 15.5 in (39.4 cm) w 17.9 in (45.5 cm) d
13 in (33 cm) h 16 in (40.6 cm) w 22.5 in (57.2 cm) d
Weight 32 lbs (13.2 kg) w/laptop
80 lbs (35.6 kg) w/laptop
50 lbs (22.7 kg) w/out monitor
80 lbs (35.6 kg) w/laptop
Sample format 1.2 & 1.5 mL Tubes 0.5 mL Tubes 96-Well Plates
1.2 & 1.5 mL Tubes 0.5 mL Tubes 96-Well Plates
ApplicationsSpecialty
Guava ViaCount® Assay Cell counting and viability
Guava® Cell Cycle Assay G0/G1, S, G2/M phase distribution
Guava Express® Cell Surface Antibodies1 Antigen detection
2
2
Guava rapidQuant™ IgG Assays1 Mouse & human IgG quantification n/a n/a n/a
ApoptosisGuava Nexin® Assay Annexin V binding
Guava® Caspase 3/7 Assay1
Caspase 3/7 activation n/a n/a 3
Guava® Caspase 8 Assay1
Caspase 8 Activation n/a n/a 3
Guava® Multi-Caspase Assay Pan caspase
Guava® TUNEL Assay DNA fragmentation
Guava® Mitochondrial Depolarization Assay Mitochondrial membrane potential n/a n/a
Cell trackingGuava® Cell Toxicity Assay Cytotoxicity: NK, CMC, ADCC
Guava® Cell Paint Assay Target cell monitoring
Guava® Cell Growth Assay Cell proliferation: Primary cultures n/a n/a
1 requires Guava Express®, Express® Plus, or Express® Pro Software Modules. 2 Users must optimize Guava Express Cell Surface Antibodies for the PCA and PCA-96 Systems. 3 Users may transfer prepared samples to tubes for collection on the EasyCyte Mini System.
Optional softwareGuava Express® Software Module (2-color) Antigen detection; immmunophenotyping n/a n/a
Guava Express® Plus Software Module (3-color) Antigen detection, GFP, immunophenotyping, antibody quantification
n/a n/a
Guava Express® Pro Software Module (4-color) Antigen detection, GFP, immunophenotyping, antibody quantification, cell signaling
n/a n/a n/a
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