The field of regenerative medicine is expanding and the advent of induced pluripotent stem cells (iPS) is essential for stem cell research, drug discovery, and disease research. Although many iPS cells are required to advance these researches efficiently, they must be grown at the same time and under the same conditions to create healthy iPS cells.
When young iPS cells mature and differentiate into neural and myocardia cells, the live cell concentration needs to be adjusted for each protocol. Cell counting is an essential part of iPS cell culturing as it enables researchers to control live cell concentration.
One of the biggest challenges when counting iPS cells is distinguishing individual cells from colonies to obtain correct counts (Figure 1). Separating cells from an iPS colony one-by-one can damage them. During manual counting, it is difficult for researchers to accurately count the number of cells in a colony, introducing a large amount of human error.
Figure 1. A colony of iPS cells (courtesy iPS PORTAL Inc.)
Olympus’ Cell Counter model R1 features liquid lens autofocusing technology and a unique software algorithm that recognizes and counts overlapping cells (Figures 2 and 3). The Cell Counter can distinguish cells clustered in a colony from feeder-free iPS cells (Figure 4). The fast, accurate auto focus enables the model R1 to count cells in around15 seconds*. Additionally, the model R1 has a field-of-view 5X larger than a hemocytometer.
Figure 2. Focus adjustment by the liquid lens
When the model R1’s counting results are compared to those generated with a hemocytometer, the R1 provides similar results (Figure 5). The Cell Counter model R1 improves the efficiency of the cell culture process, by automating cell counting and reducing variability caused by human error.
Figure 5. Result comparison between hemocytomerter and model R1 (courtesy iPS PORTAL Inc.)
When using a hemocytometer, n=4 counts were obtained, where the average number of cells counted in four squares was regarded as n=1 count. Using two Cell Counter model R1 machines, n=4 counts were obtained respectively. This set of counting was regarded as one set, and three sets of counting were carried out. The figure summarizes the data obtained from a total of three sets of counting.
*Cell counting at less than 1 x 106 cells/ml concentration of HeLa or HL-60 cells.
The Olympus Cell Counter model R1 allows for the accurate recognition of multiple overlapping cells as individual cells and counts them with a unique software algorithm that recognizes cells—in a similar manner as the human eye—and with a speedy, accurate auto focusing liquid lens technology.
Results are easily viewed on a large touch screen and data can be exported. The user-friendly design provides excellent portability and the software has robust reporting features.
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