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Guava easyCyte Systems Expanding the potential of flow cytometry for more than 20 years

20 Years of Flow CytometryExperience.Now Better Than EverMore than 20 years ago, Guava Technologies introduced thefirst compact benchtop flow cytometers. Today, the easyCyte line offers additional flexibility, greater sensitivity, and highthroughput capabilities. Powered by intuitive software,easyCyte flow cytometers are some of the most dynamic andflexible benchtop systems available.Guava - Patented Microcapillary SystemThe easyCyte line of flow cytometers offer: Up to 3 lasers and 14 parameters on a benchtop instrument,allowing for a high degree of flexibility System upgradability, which enables future-proofing An innovative microcapillary fluidics design, eliminatingsheath fluid and reducing waste stream The combination of microcapillary technology and a positivedisplacement syringe pump, which allows for direct absolutecounting with industry-leading precision An intuitive software interface, enabling a simplifiedassessment of results, including cell-health assays Detection of particles as small as 0.2 and up to 60 µm,facilitating the evaluation of a broad variety of samples High-throughput options, which allow walkaway acquisitionfor up to 96 samplesFirst Benchtop Flow 12013EasyCyte MiniBlue Laser introducedPCA-96introducedEasyCyte (Plus)introduced2006easyCyte 3rd generationHT and SL introducedeasyCyte 1st generationHT and SL introduced20102020easyCyte 4thgeneration introducedeasyCyte 3rd generation green lasersystems introduced2015With more than 20 years of experience, the Guava easyCyte Systems were the first benchtop cytometers to be introduced to themarket. The platform is continually upgraded to meet the ever-evolving needs of our customers.

642 nm488 nm405 nm60G RN525/3YLW583/250RED-R661/15RE D695/NIR785/70Inside the Guava easyCyte SystemsSSCBLU448/50How They WorkThe easyCyte Systems use patented microcapillary laser-basedtechnology to detect mammalian and microbial cells, particles,and beads. First, a sample of fluorescently labeled cells isaspirated into the microcapillary flow cell. Then, forward andside scatter characteristics are detected by photodiodes. Lasersand PMTs are then modulated to minimize crosstalk and ensurefluorescent detection is accurate. Finally, fluorescence emissionresulting from the excitation of fluorophores by the lasers isspectrally filtered and detected by several PMTs.The easyCyte Systems can resolve the fluorescence from up to12 fluorophores simultaneously.FSC405 nm488 nm532 nm642 (609/30)

Precise and AccurateAbsolute CountingGuava easyCyte Systems combine microcapillary flow with a precision pump to provide accurate and precise absolute counts—without the use of reference beads. Absolute counts can be done using cellular or bead samples.Using the ViaCount Reagent and module allows for the determination of live, apoptotic, and dead cells. Linear and accurate resultscan be seen across the multiple concentrations and on a variety of cell and sample types.Example Results From ViaCount Healthy CultureOvergrown CultureHeat-Killed Culture

Sensitive and SpecificSpherotech 8-color beads analyzed on the Guava easyCyte Systems demonstrate the instrument’sproficiency for resolving adjacent fluorophores in multiple detection channels.Plot P04, gated on P01. R1FITC MESF: 75FITCMESF:PEMESF:25 25140100CountCount140Plot P03, gated on P01. R1704010070401010100101102103104105100Green-B Fluor. (GRN-B-ALog)Green Fluorescence - 488 nm laser104105PE-Cy7 MESF: 120140100CountCount103Plot P06, gated on P01R1Plot P05, gated on P01R1701007040401010100101102103104100105Red Fluorescence - 488 nm laser102103104105NIR Fluorescence - 488 nm laserPlot P07, ungatedPlot P02, gated on P01.R1APC MESF: 50140101Near IR-B Fluo.a (NIR-B-ALog)Red - B Fluores (Red-B-ALog)Cascade Blue MESF: 40140100CountCount102Yellow Fluorescence - 488 nm laserPE-Cy5 MESF: 120140101Yellow-B Fluor.a (YEL-B-ALog)701007040401010100101102103104105RED-R Fluores. (RED-R-ALog)Red Fluorescence - 642 nm laserSmall Particle DetectionThe easyCyte Systems have been shown to detect particles assmall as 0.2 µm—a significant improvement over typical flowcytometers. This increased resolution and sensitivity allowsfor better separation, making gating and identification of dimpopulations easier. These capabilities may prove particularly usefulfor researchers analyzing particulates, beads, bacteria, and algae.Acquisition of a mixture of beads of known size demonstratesthe ability of easyCyte Instruments to detect and discriminateparticles as small as 0.2 µm.100101102103104Blue - V Fluores (BLU-V-ALog)Blue Fluorescence - 405 nm laser105

Immunological Phenotyping10 µL of adult human blood was stained for 20 minutes at room temperature with a cocktail containing anti-CD45 PerCP-Cy5.5,anti-CD3 Brilliant Violet 421, anti-CD4 Brilliant Violet 570 , anti-CD8 PE-Cy7, anti-CD16 CD56 PE, anti-CD19 Brilliant Violet510 , anti-CD45 RA APC, and anti-CD62L FITC. After incubation, cells were lysed and fixed with 180 µL of Guava Lysing Solutionfor 15 minutes at room temperature. Samples were then acquired on the Guava easyCyte 12HT System. Lymphocytes identified asCD45 were selected and subsequently gated into an SSC vs. CD3 plot. T cells (CD3 and CD45 ) were gated into a CD4 vs. CD8plot. CD4 and CD8 T cells were subtyped by evaluating each population using CD45RA and CD62L to differentiate naive frommemory cells, and to distinguish between natural killer (NK) and B cells, CD3-negative cells were gated into a plot comparing CD19(B cells) and CD16 /CD56 (NK cells).CD45 CD3CD4 CD3 CD8

SoftwareGuavaSoft operating system software provides access to modules for acquisition and analysis, as well as instrument setup andmaintenance. Modules can be used with a wide range of Luminex flow cytometry kits to simplify your flow cytometry workflows.Additionally, the GuavaSoft package includes InCyte , an intuitive open software package for custom analysis. Results can beexported to spreadsheets or as industry-standard FCS 3.0 files for further analysis. GuavaSoft Software also provides 21 CFR Part11-enabling features.InCyte Software: IntuitiveInCyte Software has an intuitive, easy-to-use interface that enables you to focus on data at the sample or experimental level.The software simplifies setup and analysis of plots with drag-and-drop features, while automated compensation makes it easyto perform complex, multi-color assays. What’s more, the Instant Update feature responds in real-time to changes in analysisconditions for viewing, and the multiparameter heat mapping function allows for analysis of entire plates of data in the timepreviously required to analyze a single sample. These features provide a simple and rapid way to attain a macroscopic view ofexperiment “hits,” and easily compare different experiments in real-time. InCyte Software is especially useful for interpreting theresults of high-throughput, cell-based assays.345672819101.Create and apply analysis methods across multipledata sets.5.2.Perform compensation during acquisition oranalysis, or use automated compensation andgain-independent compensation features.7.3.View up to 24 plots at once.4. Flow detection on automated systems.Drag-and-drop gating.6. Refine gates in real-time.Creation of statistics by default.8. Multiple gating options.9.Minimal gain adjustment needed when performingroutine assays.10. Analyzes both tubes and plates.

InCyte Software Advanced FeaturesHeat Mapping Allows for Simultaneous Evaluation of Up to 6 ParametersInCyte’s Heat Mapping feature allows the user to analyze data at the experiment level. Comparing population percentages, intensityvalues, or cell concentrations can give the user a quick, high-level view.In the example below, HeLa cells in microtiter plates were treated with various cytotoxic compounds for 24 hours. Cells were stainedusing Luminex’s MitoDamage and Caspase 3/7 kits, as well as a marker to evaluate mitochondrial stress. The InCyte Heat Mapfunction facilitated the rapid identification of compounds that showed positive results by simultaneous comparison of all 5 parameters,as shown in the pie charts below. The data shows the results for cells treated with 80 different compounds in a single exinV-CF488ECaspase3/7F7-AADGH020%20 40%40 60%60 80%Heat map allows for combination ofgroups of data to construct comparisons80 100%IC50 or EC50 Determination Within InCyte SoftwareIC50 and EC50 determinations prove critical in drug discovery, and often need advanced third party software for determination. InCyteincludes these powerful features within the software.IC50 determination using the Guava Cytochrome c Kit was analyzed with the built-in IC50/EC50 curve-fitting feature of InCyteSoftware. Cells were acquired on the Guava easyCyte 8HT System. Plot A shows the drag-and-drop gating strategy used for the IC50determination, Plot B shows the IC50 curve results for gambogic acid, and Plot C shows the IC50 for etoposide. The once complextask of generating the IC50 or EC50 curve for a given compound is automated by InCyte, based on quantitation of fluorescent signal.11 0C.IC50 38µMResponse Level, %10-1100101Concentration102103IC50 4.9µM0 1 0 2 0 3 0 4 0 5 0 6 0 70 80 9 0Response Level, %B.0 1 0 2 0 3 0 4 0 5 0 6 0 70 80 9 011 0A.10-1100101Concentration102103

Robotic IntegrationIntegrate Your Guava easyCyte with an External Robotic Arm or Liquid HandlerGuavaLink allows robot-scheduling software to control and integrate Guava easyCyte Systems with all physically compatible roboticarms and liquid-handling systems—resulting in streamlined workflows and enhanced productivity.

Guava easyCyte SL SystemsGuava easyCyte SL SystemPart NumberNumber of Channels55 055681011121314xxxxViolet (405 nm) LaserBlue (488 nm) 50 mW LaserxxBlue (488 nm) 150 mW LaserxxxGreen (532 nm) 100 mW LaserxxxRed (642 nm) 100 mW LaserxxFSCxxxxxSSC (488/16 nm)xxxxxxxxxxxxxxxxxxBlue-V (450/45 nm)xxxGreen-V (525/30 nm)xx512/18Yellow-V (583/26 nm)xx575/25Red-V (695/50 nm)xxNIR-V (785/70 nm)xxOrange-V (620/52 nm)xGreen-B (525/30 nm)xxxx512/18x512/18x512/18Yellow-B (583/26 nm)xxxx575/25x575/25x575/25Red-B (695/50 nm)xxxxxxxxxxxxxxxNIR-B (785/70 nm)Yellow-G (583/26 nm)575/25575/25575/25Orange-G (620/52 nm)x609/30xRed-G (695/50 nm)xxxNIR-G (785/70 nm)xxxRed-R (661/15 nm)xNIR-R (785/70 nm)xxxxxxMicrocapillary FluidicsxxxxxxxxxDirect, Absolute Cell CountsxxxxxxxxxDell LaptopxxxxxxxxxInCyte SoftwarexxxxxxxxxDigital Signal ProcessingxxxxxxxxxAutomation Plate and TubesMixing

Guava easyCyte HT SystemsGuava easyCyte HT SystemPart NumberNumber of Channels5HT5HT HPL6HT-2L8HTBG HT11HTBGR HT12HTBGV xViolet (405 nm) LaserBlue (488 nm) 50 mW LaserxxBlue (488 nm) 150 mW LaserxxxGreen (532 nm) 100 mW LaserxxxRed (642 nm) 100 mW LaserxxFSCxxxxxSSC (488/16 nm)xxxxxxxxxxxxxxxxxxBlue-V (450/45 nm)xxxGreen-V (525/30 nm)xx512/18Yellow-V (583/26 nm)xx575/25Red-V (695/50 nm)xxNIR-V (785/70 nm)xxOrange-V (620/52 nm)xGreen-B (525/30 nm)xxxx512/18x512/18x512/18Yellow-B (583/26 nm)xxxx575/25x575/25x575/25Red-B (695/50 nm)xxxxxxxxxxxxxxxNIR-B (785/70 nm)Yellow-G (583/26 nm)575/25575/25575/25Orange-G (620/52 nm)x609/30xRed-G (695/50 nm)xxxNIR-G (785/70 nm)xxxRed-R (661/15 nm)xNIR-R (785/70 nm)xxxxxxMicrocapillary FluidicsxxxxxxxxxDirect, Absolute Cell CountsxxxxxxxxxAutomation Plate and TubesxxxxxxxxxMixingxxxxxxxxxDell LaptopxxxxxxxxxInCyte SoftwarexxxxxxxxxDigital Signal Processingxxxxxxxxx