细胞计数法(国外英文资料).docVIP

细胞计数法(国外英文资料)Cell counting methodCell counting is a method used to count the number of cells in a cells suspension. Normally use the counting board (blood ball counting board). Can be used to separate (powder) cell culture inoculation before counting the number of cells in cell suspension preparation by, also can be used to culture cell number count. The dispersed cell suspension is required, regardless of the object of the count.The preparation of cell suspensionFor the cells in suspension, the following steps 2 (counting and counting) are performed directly. If the counting object is a cell that grows on the wall, the first thing you need to do is make a cell suspension.1), end culture, suck the culture medium out, use PBS to wash the culture material once.2), 1 ml to join in the culture bottle, 0.25% trypsin solution in 37 ℃ digest 1-3 min. Watch under the mirror. As the cells get round to the wall, the digestive juices are discarded.3), add a certain amount of culture (if these cultured cells are no longer useful, add PBS), blow with a straw, and make the cells take off the wall and make a cell suspension.Counting and counting1) in the center of the cell counting board, a special cover slide is placed.2) use a glass siphon to absorb the cells and let the siphon flow out of the counter on the cover of the cover glass or the lower side of the cover glass, until the slide is filled with liquid.3) the total number of cells in the four corners of the large grid under the microscope. For the cell line, the cells count only on the line and the left line.4) the density of the cell suspensions in the lower count:Cell density = (4 large square cells in total / 4) x dilute multiple times 104 / mlNote: the dilution (multiply by at least 2, because of the volume mixing with trypanblue)The rule of the upper diagram counts: represents the count: not counting:Example:T75 monolayer made 10 ml of cell suspension culture, take 0.1 ml solution with 0.1 ml trypan blue mix in test tu