EXPERIMENT : CELL COUNTING FOR DETERMINING VIABLE CELL CONCENTRATION
Objectives:
- To count adherent and suspension cell using hemocytometer.
- To calculate viable cell concentration in a suspension.
Procedures:
Counting Viable Cells using Hemocytometer/Automated Cell Counter
▪ Prepare the hemocytometer slide by cleaning the surface with 70% ethanol carefully (do not scratch the semi-silvered coating). Apply the same step for the coverslip. Wet the edges very lightly and press it down over the grooves and semi- silvered counting area.
▪ Collect 20 μl samples from the cell suspension using a pipettor and transfer it immediately to the edge of the hemocytometer chamber.
▪ Expel the suspension and allow it to be drawn under the coverslip by capillary force. Do not overfill the chamber. Blot off any surplus liquid and place the slide under the microscope.
▪ Alternatively, trypan blue dye can be used to stain cells by mixing an equal volume of trypan blue to a cell suspension (1:1) to visualize and distinguish the viable and dead cells. Leave the mixture for 2 minutes at room temperature.
▪ Move the slide to focus on counted area of the grid.
▪ Move to the next chamber and perform a second count. At least 100 cells should be counted for statistical validity of the final value.
QUESTION:
From this,please make a general introduction on the experiment.
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