Key Factors For Reliable Cell Counting Results
Cell counts is a critical parameter for cell banking, process development and many basic biology biotechnology processes. Keep in mind these following factors for the reliable accurate cell counting.
- Sample cell concentration must be in the linear range specified by the particular cell counter. For single image based cell counters, 5X10^5 /ml may be the lowest concentration you can work with. This is much determined by the sample preparation process and sampling statistics. For example, 10ul cell sample, add 10ul Trypan Blue, take 10 ul for counting, image captured by the cell counter camera is around 0.2uL to 0.4uL. To convert back the concentration from the image to concentration per mL, any statistical errors inherited from the original image counting will be amplified by at least 10^4. Therefore use the recommended sample concentration will ensure the final counts reliable and statistically meaningfull.
- Sample and dye mixed well, cells distributed evenly in the counting chamber. Minimize pipetting errors, let the loaded counting slide rest on a solid even surface for 15 to 20 seconds, avoid hold the counting slides side ways or tilting to one side before insert the slide into cell counter.
- Adjust focus on your cell counter. Off focus will result inaccurate cell sizes and viability.
- For fluorescent counter, adjust exposure of green and red channel for better results. Over exposure or under exposure will result inaccurate viability. You want a bright image with no background noises for both channels.