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Cell counting is a routine task in cell biology and molecular biology. Counting results contribute significantly to assay accuracy. However, currently available automatic cell counting methods often require costly instruments with liquid handling and cleaning hardware. Daily usage requires special reagent and sample preparation, along with routine calibration and maintenance. Due to these limitations, cell counting remains largely a traditional manual operation with hemacytomers for most research laboratories.
Cellometer™ Auto T4 is an automated system to replace manual cell counting. Like using a hemacytometer, it requires no special counting buffer or cell manipulation. Cells in growth media, PBS or other solutions can be pipetted directly into disposable counting chambers. For cell viability, the standard trypan blue method is used. By using disposable, all plastic counting chambers, samples are isolated within each counting chamber, thus eliminating needs for liquid handling hardware and risk of cross contamination by different samples. There is no washing between each sample and no chance to clog the system.
Materials and MethodsCellometer™ Auto T4 acquires cell data from multiple locations of counting chambers. The system includes a compact instrument with advanced imaging-based cell counting software, which automatically analyzes acquired cell images and measures cell concentration, viability, and cell size. Each type of cell uses a set of cell type parameters, which are established and saved for repeated use. For a new cell type, parameters can be imported from a library of cell type parameters, or they can be established automatically using a “New Cell Type Wizard”. Once set up, switching from one cell type to another only involves the selection from a drop down menu.
For data show below, suspension cells were directly pipetted into Cellometer™ disposable cell counting chamber. For attachment cells, they were trypsinized prior to loading into a counting chamber. For cell suspension diluted prior to loading into counting chamber, a dilution factor was applied for the software to directly convert cell concentration results to reflect the dilution.
Cell counting and analysis procedure consists of a few simple steps: 1. Pipette 16 to 20 µl of cell suspension into a disposable cell counting chamber; 2. Insert the loaded chamber into the instrument; 3. Inspect cell morphology and determine cell concentration, viability and cell size automatically; and 4. Save cell counting data, cell images and histogram.
Results and ConclusionsSome representative data are shown in Figures 1 and 2. Cellometer™ Auto T4 proves to be an effective method to automatically count cells, determine viability, and measure and record cell size and distribution.
