Nexcelom - Model Cellometer K2 - Fluorescent Cell Counter
From Fluorescent Automated Cell Counters
The Cellometer K2, powered by Matrix software, utilizes brightfield imaging and dual-fluorescence imaging to quickly and accurately identify and count individual cells. Cell count, concentration, diameter, and % viability are automatically calculated and reported.
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Features
The Cellometer K2 has the following advantages:
- Dual fluorescence and brightfield imaging – stain only nucleated cells for the most accurate count and viability information
- Fast results – count, size, concentration, and viability calculations in
- Analyze complex samples – designed for analysis of complex and messy samples including whole blood, peripheral blood, cord blood, and bone marrow
- Multiple fields of view – increased accuracy with the ability to capture one, four, or eight images per sample
- Built-in predefined assays – quickly analyze viability, apoptosis, and transfection efficiency
- Built-in cell types – includes saved parameters for over 400 cell types
- Small sample volume – only 10 µl of cell sample required
- Customizable reports – includes predefined reports with the ability to create new ones with graphs, images, charts, and tables
- Multi-language support – over 7,000 languages available
- 21 CFR Part 11 ready – optional add-on that includes an audit trail, user access control, and digital signature
The Cellometer K2 can be customized to handle a variety of cell types, including primary cells, tumor digest, insect cells, cell lines, fragile cells, and more at low or high concentrations.
Take the guesswork out of setting up your cell quantification experiments. The Cellometer K2 comes with frequently used assays and cell types with predefined settings to ensure consistent results from sample to sample. Easily build custom assays and cell types to fit your experimental needs.
Not sure what settings to use? Our customer success team and field application specialists are here to help you develop fit-for-purpose assays and protocols for your specific research and development requirements.
Capture one, four, or eight images per sample. The instrument default is set to four images, which is the equivalent of six quadrants on a hemocytometer. Eight images are equivalent to twelve quadrants on a hemocytometer. The ability to capture multiple images improves the cell counting dynamic range and accuracy of results.
Dual-fluorescent Staining for Clumpy CellsThe fluorescent image (far right) shows bright green AO-positive hepatocytes declustered by the Cellometer K2 algorithm. Red circled hepatocytes are PI-positive (dead) while free nuclei are not counted.
Use default or customized reports based on the data and images you want to be included for your specific experimental needs. Automatically export images and data reports including CSV, Excel, Word, or PDF files. Perform statistical analysis for a wide range of parameters such as average, variance, min/max, and standard deviation of cell size.
21 CFR Part 11 ReadyAn optional module can be purchased to meet the requirements for 21 CFR Part 11. The additional module comes with the following features:
- User login with passwords
- User assigned permissions
- Audit trail
- Error log files
- Electronic signatures
Because brightfield cell counting does not differentiate nucleated from non-nucleated cells and trypan blue staining is not as easy to detect as fluorescent staining, dual-color fluorescence is strongly recommended for accurate viability analysis for primary cells. The K2 is equipped with standard assays for dual-fluorescence analysis of a variety of cells stained with Acridine Orange and Propidium Iodide (AO/PI).
The AO/PI MethodAcridine Orange (AO) is a nuclear staining (nucleic acid binding) dye permeable to both live and dead cells. It stains all nucleated cells to generate green fluorescence. Propidium Iodide (PI) can only enter dead cells with compromised membranes. It stains all dead nucleated cells to generate red fluorescence. Cells stained with both AO and PI fluoresce red due to quenching, so all live nucleated cells fluoresce green and all dead nucleated cells fluoresce red.
No Interference from Red Blood Cells, Platelets, or DebrisThe dual-fluorescence AO/PI method utilizes nuclear staining dyes that bind to nucleic acids in the cell nucleus. Because most mature mammalian red blood cells do not contain nuclei, only live and dead mononuclear cells produce a fluorescent signal. There is no need to lyse red blood cells, saving time and eliminating an extra sample preparation step. Red blood cells, platelets, and debris are not counted in the fluorescent channels.
NCI-60 is a group of 59 human cancer cell lines (originally 60) developed by the National Cancer Institute for screening purposes.
- 57% of the NCI-60 cell lines are clumpy, contain debris, or display large variations in cell shape or size
- All 59 NCI-60 cell lines have been successfully validated on the Cellometer Image Cytometer
All 40 of the NCI Comprehensive Cancer Centers use Cellometer Cell Counters.
The MCF-7 breast cancer cell line can be very clumpy. The Cellometer pattern-recognition software identifies and counts individual cells within these cell clumps for accurate analysis (shown above).
The Cellometer cell roundness setting can be adjusted for recognition and counting of irregular-shaped cells, such as RD cells and activated T-cells.
Counting 1 x 106 cells takes approximately 5 minutes with a manual hemacytometer. Counting live and dead cells sometimes takes twice as long. The Cellometer K2 Image Cytometer calculates cell count and concentration for live and dead cells and % viability in just 60 seconds.
- Eliminate wash steps
- Eliminate judgment errors
- Eliminate interference from RBCs
- Eliminate recording & calculation errors
- Reduce counting time … run more experiments
We offer two different disposable counting slides, one with protective coverings on both sides and a ready-to-use option packed in microscope slide boxes.
- No clogging
- Time savings — no washing
- No risk of cross-contamination
- Reduce biohazard risk to users
Applications
Measure live cell concentration and viability without lysing red blood cells for consistent results from patient samples. »
Rapidly identify fluorescence-positive cells from a sample, calculate cell concentration, size, and determine the GFP transfection percentage automatically. »
Measure nucleated cell concentration without lysing red blood cells using nuclear staining dyes (AO), for human and mouse blood. »
Quantify cell viability and concentration for a variety of immunologically relevant samples such as: bone marrow, cord blood, splenocytes, lymphocytes, isolated mononuclear cells, tumor digests, murine samples, and others. »
Measure live cell concentration and viability for baculovirus infected insect cells. »
Measure live hepatocyte concentration and viability from fresh and cryo preserved samples using dual-fluorescent nuclear stains for human, rat, mouse, and horse. »
Measure live cell concentration and viability of cancer cell lines used in oncology research and biology research. »
Perform cell based assays and measure cell size, viability, and concentration of cell lines and primary samples used in adoptive cell therapy research. »
Detect and analyze apoptotic and necrotic cells with Annexin V and PI. »
When should you use trypan blue and when should you use acridine orange/propidium iodide to measure cell viability? »
Cell viability is performed using various fluorescent membrane exclusion dyes, such as PI, EB, 7AAD, and others. This assay is performed by enumerating cells in captured bright-field and fluorescent images. And Necrotic cells are detected using propidium iodide. »
Analyze fresh and processed blood and bone marrow samples without lysing: no interference from RBCs. »
Performing cell size measurement assay and using cell size to count cells within preset cell size parameters. For adipocytes, stem cells, Sf9 cells, dendritic cells, and others. »
Specifications
Includes
- Cellometer K2 Instrument
- Cellometer Software
- Two Standard Fluorescence Optics Module
- Power Supply
- USB 2.0 Connection Cable
- Phone/online applications support during set-up
Available Accessories
- PC Laptop
Imaging Performance
Cell Size: 4-90 microns
Conc. Range: 105 – 107 cells/ml
Brightfield imaging, fluorescent imaging and pattern-recognition software to quickly and accurately decluster, identify and count individual cells.
Instrument Specifications
Weight: 23.0 lbs. (10.4 kg)
Dimensions: Width: 6.0” (15.2 cm), Depth: 8.5” (21.6 cm), Height: 14.0” (35.6 cm)
Input to Power Adapter: 100-240 VAC, 50/60 Hz, 1.0A
Output to Instrument: 12 VDC, 3.34A
PC / Laptop Minimum Requirements: (If purchasing Cellometer without PC laptop)
- Windows 10, 64-bit
- Intel® Core i7, 2.0 GHz processor
- 16GB RAM (or higher)
- NVIDIA P1000 GPU (equivalent or better)
Available Fluorescence Optics Modules
Excitation / Emission: 470nm/535nm
Example Fluorophores:
- Acridine Orange
- CFDA
- FITC
Excitation / Emission: 540nm/660nm
Example Fluorophores:
- Propidium Iodide (PI)
- Ethidium Bromide
- 7-AAD
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