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1. There is a total of 56 cells including 20 unstained cells altogether in the five counting squares of hemocytometer.Given volume of media is 5ml. The average viable cell count per square is 4 and the dilution factor is 2. Then calculate the percentage of cell viability, viable cells/ml and the volume of media needed, if we need 5000 cells.

2. What is the principle behind Trypan Blue Exclusion Test of cell viability?

3. If you are provided with a cell sample, how will you be able to find out its concentration by using a hemocytometer?

4. Which are the different dyes used to differentiate between viable and non viable cells?

5. Explain the different cell counting techniques available other than Hemocytometer.

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Expert Solution

2) The principle for the trypan blue exclusion test is that the live cells possess intact cell membranes which can exclude many types of dyes, such as trypan blue, eosin etc and most importantly dead cells do not.This test is used to know the number of viable cells present in a cell suspension.

3) The counting of cells is much easier with a hemocytometer as it contains a thick glass microscope slide with a number of perpendicular lines. It has specified dimensions so, the area covered by the lines can be easily known which makes it possible to count the number of cells in a particular solution.

4) The different types of dyes used to differentiate are fluorochrome dye; as it helps in making easier to identify the viable cells. And also various vital dyes such as trypan blue, methylene blue are used.

5) TC20 Automated cell counter; dye exclusion, clonogenic assay; and the three trypan blue exclusion based methods are manual, semi automated and automated.

gladiator answered 2 years ago

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