Questions about cell counting A.What if a cell is on the lines that define a big square”? Do you count it? The general rule of thumb is that ifmore than 50% of the cell is iside of the big square most important thing is to be consistent. For example, if you and your group decide that in order for counted, it should be completely inside the first big square, make thing for the other four big squares you should count it. The sure you and your group members do the same B. What if you see a large “clump of cells? How do you count i? PIK2 cells produce a lot of extra cellular matrix and sure your cell pellet is completely re-suspended before cou your cell pellet, try your best to estimate the number of cells in a down more in future experiments. can clump together. This is why it is very important to make ed in solution before counting the cells. If you did not fully re-suspend clump and make sure to pipet the cells up and Practice Problem 1- Counting Cells: Using the equations described on the previous page to determine the concentration of cells (cells/ml), total cells in cell suspension, percent viable cells and percent nonviable cells The following hemocytometer grid shows PtK2 cells that were re-suspended in 4mL of media. Solve for the four equations above (C-F) and SHOW YOUR WORK FOR EACH o Which cells are represented by this open circle? B. Which cells are represented by this dark circle? C. Total cells/mL 0 D. Total cells in suspension . E. Percent Viable Cells- 0 F. Percent Non-viable cells-