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Talk:Cell count and normalization in HRR

From Bioblast
Revision as of 07:57, 25 September 2020 by Schmitt Sabine (talk | contribs)

Feedback from Sabine (email 2020-09-23) General I think it should be pointed out that Nce refers to the total cell count, i.e. the sum of viable cells (Nvce) and dead cells (Ndce). I know this is explained here https://wiki.oroboros.at/index.php/Living_cells#The_units_of_the_cell_count_and_cell-count_concentration, so maybe this link should be implemented. In my experiments I always normalized to the number of viable cells, which I think is also an option. No matter whether the Nce or Nvce is used, the viability should be >95% and comparable between samples. Therefore I would suggest to include the viability in the excel sheet. Not for calculation but for documentation.

Complete replacement β€œThe cell-count concentration Cce calculated from the initially determined Cce,Οƒ (3) can be compared with the median of the finally determined cell-count concentrations Cce.i (13) for quality control of cell counting.” In my understanding, this is not (only) a quality control of cell counting but a quality control of proper dilution.

Pipetting into V#-VJ.i Why are no subsamples taken from the chamber? (Cell count concentration control and further assays) I think a control of the cell count concentration in the experimental chamber would be important due to the higher dilution factor (compared to the complete replacement) and the removal of medium with the volume Vj,I which bears another risk of error due to the additional pipetting step.

Pipetting into V+VStopper capillary Same as before: Why are no subsamples taken from the chamber? (Cell count concentration control and further assays) excess volume: VStopper capillary + sampling + waste total volume before subsampling from chamber and closing chamber

This implies that subsamples are taken from the chamber but it is not mentioned in the text or shown in the figure. I think it could be easily implemented as the total volume V# is 2.59 mL before closing the chamber.