Abstract
Cell counting has become an essential method for monitoring the viability and proliferation of cells. A hemacytometer is the standard device used to measure cell numbers in most laboratories which are typically automated to increase throughput. The principle of both manual and automated hemacytometers is to calculate cell numbers with a fixed volume within a set measurement range (105 ~ 106 cells/ml). If the cell concentration of the unknown sample is outside the range of the hemacytometer, the sample must be prepared again by increasing or decreasing the cell concentration. We have developed a new hemacytometer that has a multi-volume chamber with 4 different depths containing different volumes (0.1, 0.2, 0.4, 0.8 µl respectively). A multi-volume hemacytometer can measure cell concentration with a maximum of 106 cells/ml to a minimum of 5 × 103 cells/ml. Compared to a typical hemacytometer with a fixed volume of 0.1 µl, the minimum measurable cell concentration of 5 × 103 cells/ml on the multi-volume hemacytometer is twenty times lower. Additionally, the Multi-Volume Cell Counting model (cell concentration calculation with the slope value of cell number in multi-chambers) showed a wide measurement range (5 × 103 ~ 1 × 106 cells/ml) while reducing total cell counting numbers by 62.5% compared to a large volume (0.8 µl-chamber) hemacytometer.
Highlights
Cell counting has become an essential method for monitoring the viability and proliferation of cells
In the conventional Fixed-Volume Cell Counting (FVCC), the cell concentrations were calculated by counting cells in each chamber at different volumes
The number of cells increases as the chamber volume increases, i.e. at fixed volume 0.1 μl, there are 10 ~ 500 cells at 105 ~ 5 × 106 cells/ml while at fixed volumes 0.2 μl, there are 10 ~ 500 cells at 5 × 1 04 ~ 2.5 × 106 cells/ml
Summary
Cell counting has become an essential method for monitoring the viability and proliferation of cells. A hemacytometer is made from optical glass for use under a microscope and consists of 2 parts: a thick glass slide and cover glass with a small gap to contain cell suspension in a 0.1 μl small grid area size 1 × 1 mm[8] It cannot be accurately measured in high concentrations because the cells too crowned and difficult to count. The digital camera captures cells on the slide in the fixed volume and analyzes them using specialized software for cell counters to calculate cell concentration based on the fixed volume[6] Both manual and automated hemacytometers use a fixed volume to measure cell concentration. Though manual and automated hemacytometers are standard in most laboratories, they cannot measure low cell concentration samples and have a narrow measurement range (105 ~ 106 cells/ml) due to the fixed volume containing cell.
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