counting cells before adding them to 96 well plate
Posted 23 August 2012 - 07:12 PM
can any one please help explaining the hemocytometer and specially the calculation part, AM REALLY LOST.
am using HepG2 cell line which is cultured in petridish and I need to have 5000 cell/ well .
Posted 23 August 2012 - 07:31 PM
There should be a grid on the hemocytometer that you see through the microscope, with some bigger and smaller squares. Each square represents both a unit of area and a unit of liquid volume (because the specialized coverslip you put on it has a specific weight to it, in which only a specific amount of liquid can be under it). Based on the number cells per square, or set number of squares, you can calculate the number of cells you have in your sample, and from that calculate the volume needed to give you 5000 cells.
What I would do is find the brand of hemocytometer you have, go online and find the data sheet for it. That should tell you the units of area and volume that each square on the grid represent.
Does that help/make sense?
Posted 24 August 2012 - 10:35 PM
I have another question, I did count my cells and I had total # 103 cells from 4 squares.
now, i need to have 5000 cells per well (96 well plate) to do MTS assay.
I kind of lost now in the calculation part.
concentration (cells/ml)= 103/4 x 2 (dilution factor) x 10,000 = 515000
to calculate total number of cells= conc. X vol. of sample ( I did not understand which vol.) ???
after that what I should do exactly to get the number of cells I want ??
Posted 26 August 2012 - 01:05 AM
Posted 04 September 2012 - 06:55 AM
Posted 28 November 2012 - 06:38 PM