Q: I want to know what the dilution factor is when using a blood cell counting plate. For example, when nine drops of cell solution and one drop of trypan blue are mixed and then dropped on the counting plate, the dilution factor is 10 Right, blood cell counting plate counting. Many literatures say that add 1-2 drops of stained cell suspension to the edge of the cover glass to completely fill the space between the counting plate and the cover glass. I do n’t understand how to add the cell suspension.
Answer: 1. After blowing the cells into a single-cell suspension, just place the dropper against the gap between the counting plate and the cover glass, and apply light pressure, because the negative pressure cell suspension will easily enter the cover glass and Between counting boards. But be careful not to blow too much cell suspension, this will lead to inaccurate counting, as long as the gap is filled, there should be no air bubbles. (There are counting areas on both sides of the counting plate, when adding the cell liquid, add one side. Yes, the other side is spare.)
2. Generally count the four large squares on the side (16 small squares each), take the average, and multiply by 10 to the fourth power. This is the number of cells per milliliter. Multiply by the amount of your cell fluid, for example, 4 ml, is multiplied by 4.
"Multiplied by the amount of your cell fluid" refers to the original total amount of cell suspension waiting to be counted.
What we add to the counting plate is just a small amount of sample taken from the original cell suspension waiting to be counted for the purpose of detecting the concentration.
Add the solution of the counting plate and flatten the counting plate.
You can see: The formula for counting the concentration of cell suspension is
Cell concentration = (total number of 4 large cells / 4) × 104 cells / ml
Use this result (count the technetium cell concentration on the counting plate) × the total amount of your cell suspension = the total number of cells in the cell suspension waiting to be counted.
The definition of dilution factor is how many times the concentration before dilution is the concentration after dilution?
1. A drop of cell liquid plus a drop of staining solution, the concentration before dilution is 100%, the concentration after dilution is 50%, the dilution factor is 2
2. 9 drops of cell solution plus one drop of staining solution, the concentration before dilution is still 100%, the concentration after dilution is 9/10, the dilution factor is 10/9
The dilution factor is based on the conventional mathematical understanding. It is recommended that you use a hemocytometer to count on the sides of the plate with alcohol and then cover it with a cover glass, so that the cover glass is attached more firmly. Correctness helps, many foreign laboratories do this !! Then it is best to have a fixed volume when counting, generally 7--9ul is better, using a micro sampler is the best !!
If the number of cells is large, one drop (or 100 ul) of cell suspension + eight drops (or 800 ul) of cell culture fluid + one drop (or 100 ul) of moss blue can be taken, so that the final dilution factor is ten times.
The dilution factor is the factor that you use to dilute the cell suspension you finally get. For example, if you digest a bottle of cells and make a suspension and want to divide it into bottles or inoculate, you need to count it. Drop it into the ep tube and add another drop of placenta orchid plus eight drops of buffer or normal saline. Then you get a ten-fold dilution of your cell suspension. You can read this value directly with the formula. Of course, the 10 in the formula is the dilution multiple. If you still do n’t understand it, you can understand it. You will understand it after doing it several times.
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