Cellometer Bright Field Cell Counters
- Cell count and trypan blue viability in <10 seconds!
- Proprietary Cell Membrane Outline Algorithm to count irregular shaped cells, cells in clusters and produce cell size histograms
- View, print, and save counted cell images, data reports, and cell size histograms
Find the Cellometer Cell Counter for Your Lab
Cellometer Auto 1000
A convenient stand-alone system with a large, userfriendly touch screen.
Cellometer Auto T4
Used by all 40 NCI comprehensive cancer centers and the top 10 pharmaceutical companies.
The most affordable option for accurate, automated cell counting and trypan blue viability.
Examples of Cellometer in Publications
Cell count for large cancer cell line panels: “A genome-wide screen for microdeletions reveals disruption of polarity complex genes in diverse human cancers.” Rothenberg SM, Mohapatra G, Rivera MN, et al. (2010) Cancer Res 70: 2158-64.
Cell viability by trypan blue: “Increased Expression of the dsRNA-Activated Protein Kinase PKR in Breast Cancer Promotes Sensitivity to Doxorubicin.” Bennett RL, Carruthers AL, Hui T, et al. (2012) PLoS One 7: 1-12.
Cell size analysis for mesenchymal stem cells: “Quantitative Approaches to Detect Donor and Passage Differences in Adipogenic Potential and Clonogenicity in Human Bone Marrow-Derived Mesenchymal Stem Cells.” Lo Surdo J, Bauer SR. (2012) Tissue Eng Part C Methods 18: 877-89.
AllCells is excited to announce our collaboration with Nexcelom, the clear leader and innovator in the field of image-based cytometry for cell analysis.
At AllCells,we understand the importance of continually updating our research instruments with the most innovative in the field in order to provide our customers with the highest quality and technologically advanced products and services. Late last year, we began using Nexcelom’s Cellometer technology to more accurately count our cells in lab. (more…)
Perform image-based GFP analysis WITHOUT all of the set-up, maintenance, and data manipulation required for flow.
Measure GFP Expression Efficiency Using Image Cytometry —
In just a few clicks, Cellometer Vision CBA generates brightfield and fluorescent cell images, and detailed data reports.
- Detection of GFP expression to quantify the transfection/transduction efficiency.
- Single assay for measuring GFP expression and viability
- Measurement of other fluorescent proteins
Detection of GFP Expression to Quantify the Transfection/Transduction Efficiency
- Load only 20 µl sample into counting chamber
- Capture and analyze brightfield and fluorescent cell images automatically
- Data read-out includes cell counts, concentrations, and percentage of GFP positive cells
Single Assay for Measuring GFP Expression and Viability in Mouse Embryonic Stem Cells
- Stain Sample with propiduim iodide
- Monitor the viability of the culture during GFP expression
- Determine the viability of GFP expressing cells
Measurement of Other Fluorescent Proteins
Cellometer can measure other fluorescent proteins including dared, YFP, CFP, tdTomato and Saphire.
Advantages of Image Cytometry:
- No washing. No clogging. No daily calibration
- Bright field and fluorescent cell images for verification of counted cells
- Simple procedure with pre-defined instrument settings and data output
- Small, 20µl sample size
- Non-fluidic instrument … maintenance-free
Autophagy is an important cellular catabolic process that plays a variety of important roles, including maintenance of the amino acid pool during starvation, recycling of damaged proteins and organelles, and clearance of intracellular microbes. First, the damaged proteins, organelles or foreign microbes are isolated by double membrane vesicles called autophagosomes. The vesicles then complete the enclosure of the damaged organelles and then fuses with lysosome. to form autolysosomes. Finally, the materials are degraded within the autolysosomes and the nutrients are recycled back to the cell. Currently employed autophagy detection methods include fluorescence microscopy, biochemical measurement, SDS-PAGE, and Western blotting, but they are time-consuming, labor-intensive, and require much experience for accurate interpretation.
By utilizing image cytometry, fluorescently labeled autophagosomes can be detected and measured via fluorescence analysis. Cellometer Vision has been used to demonstrate the detection and measurement of total fluorescence intensity of fluorescently labeled autophagosomes using proprietary stains (Cyto-ID® Green dye from Enzo Life Sciences) and GFP-LC3 protein in live cells.
The image cytometry method has been shown to generate comparable fluorescence intensity data to flow cytometry, which can be an alternative method to indicate the autophagy activity level in live cells. This can serve as a useful method to quickly assess autophagic response of live cells when treated with various chemical or biological reagents, as well as an alternative technique available in support of autophagy-based drug discovery relating to various pathological disorders.
What is GFP?
Green Fluorescent Protein (GFP) is a 26.9 kDa protein first identified in crystal jellyfish, Aequorea victoria. It was discovered that when exposed to blue or ultraviolet light the protein fluoresces green. After GFP was first expressed in E. coli in 1994 it was soon confirmed that GFP can also be successfully expressed in other organisms as well. Since then, not only have many fluorescent proteins of different colors been generated, but their function is enhanced to provide a faster and stronger fluorescent signal.
- GFP is often used as a reporter of gene or protein expression. By detecting GFP expression it is possible to quantify the transfection/transduction efficiency.
- By staining the cells with propidium iodide we can monitor the viability of the culture during GFP expression.
- In cultures that are co-transduced with GFP and RFP, the Cellometer has the capability to capture, analyze, and report the population of GFP positive, RFP positive, or dual positive.
Acquiring GFP Expression Efficiency Using Cellometer
With the Cellometer Vision CBA, just 20µl of sample is added to the Cellometer Counting Chamber. Imaging and analysis of GFP expression is completed in less than 60 seconds. Bright field and fluorescent cell images can be viewed to check cell morphology and verify cell counting. Total cell count, concentration, and mean diameter are automatically displayed.
Quantifying GFP Efficiency in 4 Easy Steps
- Pipette 20 µl of cell sample into a disposable counting slide
- Insert slide into the instrument
- Select assay from a drop-down menu
- Click count, acquire image and view cell count, concentration, diameter and percent of GFP positive cells
Jurkat cells were used to analyze cell cycle kinetics following treatment with the cell-cycle-arresting drug etoposide. Etoposide is designed to arrest the cells at the G2 phase of the cell cycle. Jurkat cells were incubated with media only (control) or etoposide (0.06 µM, 0.12 µM) for 24 hours. Control and drug-treated cells were ethanol fixed and stained with cell cycle propidium iodide reagent. For each sample, 20 µl of cell sample (at ~4 x 106 cells / mL) was loaded into a Cellometer imaging chamber, inserted into the Vision CBA Analysis System, and imaged in both bright field and fluorescence. The fluorescence intensity for each cell was measured and a cell cycle histogram is automatically generated for each sample using the optimized Nexcelom cell cycle data layout in FCS Express 4 Flow Software. Gating can be manually optimized directly on the histogram with automatic update to the associated data table.
The Cellometer fluorescence-based cell cycle analysis can be effectively implemented in studies examining the efficacy of cell cycle arresting drugs.
One of the most common and popular methods for cell cycle detection is the use of fluorescence-based dyes. There are a number of fluorescent-based dyes that are capable of binding to double stranded DNA upon cell fixation. Propidium iodide (PI) and DAPI are two such dyes. Since the amount of bound fluorescent dye is directly proportional to the amount of DNA present within a cell, these dyes can be used to detect the cell cycle within a population of cells.
How it works:
Since the amount of DNA doubles from 2n to 4n between G1 and G2/M phases, and the amount of PI incorporated is correlated to the amount of DNA within each cell, we can generate a histogram based on PI fluorescence intensity.
The Cellometer instrument acquires a bright-field and a fluorescent image for each sample tested. The bright-field image allows researchers to verify cell morphology, evaluate the degree of homogeneity of the sample, and identify the presence of cellular debris
The fluorescent counted image can be used to confirm that cells are counted correctly. Individual counted cells are outlined in green. Uncounted cells are outlined in yellow. Cellometer software uses proprietary algorithms to accurately count individual cells within clumps.
Once counting is complete, a cell cycle histogram is automatically generated for each sample using the optimized Nexcelom cell cycle data layout in FCS Express 4 Flow Software. Gating can be manually optimized directly on the histogram with automatic update to the associated data table.
Representative data set:
In this example, untreated Jurkat cells were used to analyze cell cycle kinetics. The cells were ethanol fixed, stained with propidium iodide cell cycle reagent, and 20 µl of cell sample was loaded into a Cellometer counting chamber. The counting chamber was inserted into the Vision CBA Analysis System, and imaged in both bright field and fluorescence. The fluorescence intensity histogram and associated data table was generated in FCS Express 4 software.
Cellometer Vision incorporates image based cell counting and fluorescence detection in a compact and easy-to-use instrument. With fluorescence detection capabilities, Cellometer Vision is an ideal solution for many complex cell population characterization assays such as rapidly counting white-blood-cells in whole blood.
Acridine orange (AO) is a fluorescent nucleic acid stain that has been used to measure nucleated cell concentration in various cell types, including mammalian cell lines, mammalian primary cells, and yeasts. AO is a membrane-permeable dye that stains nucleated cells green.
The following article describes a simple and specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns.
Author institutions: Translational Immunology Section, Office of Science and Technology, National Institute of Arthritis and Musculoskeletal and Skin Diseases, and Translational Autoinflammatory Disease Section, Office of the Clinical Director, National Institute of Arthritis and Musculoskeletal and Skin Diseases
Authors: Barbara Yang, Tuyet-Hang Pham, Raphaela Goldbach-Mansky, Massimo Gadina
Journal: J Vis Exp. 2011; (49): 2662.
A key component of this assay is to standardize the dose response to the number of cells stimulated. They have found 2×106 cells /mL is the sufficient concentration for optimal and measureable concentration of IL-1beta and other cytokines.
“Using Cellometer Vision, diluted blood can be counted without lysing RBC y choosing the whole blood count option using acridine orange (AO). Add one part diluted blood with one part AO and load 20 ml of the 1:1 mixture into a disposable counting chamber. The final AO concentration after dilution should be 1mg/mL. Adjust accordingly with incomplete RPMI 1640 media to final cell centration of 2×106 cells /mL.”
Cellometer Vision has a convenient cell concentration adjustment calculator. Input the original concentration of 20mL, based on the measured cell concentration for each sample, the software calculates the adjustment required to obtain 2×106 cells /mL.
Accurate Measurement of Peripheral Blood Mononuclear Cell Concentration using Image Cytometry to Eliminate RBC-Induced Counting Errors
Peripheral blood mononuclear cells (PBMCs) have been widely researched in the fields of immunology, infectious disease, oncology, transplantation, hematological malignancy, and vaccine development. Specifically, in immunology research, PBMCs have been utilized to monitor concentration, viability, proliferation, and cytokine production from immune cells, which are critical for both clinical trials and biomedical research.
Ten Questions to Ask Before Purchasing an Automated Cell Counter
Believe it or not, all cell counters are not created equally. Ask a colleague with a handheld counter that sits in a drawer or a bench-top cell counter that acts as a paperweight and they will tell you that some cell counters don’t work as expected. Before purchasing an automated cell counter, ask the representative the following questions. Knowing these answers will help ensure that you purchase the right cell counter for your applications. (more…)