实验中细胞铺板的方法(国外英文资料).docVIP

实验中细胞铺板的方法(国外英文资料) The cell count and the planking of the test, such as invasion and migration [experimental steps] 1, prepare for work 75 percent of the alcohol is rubbed against both hands, while using an alcohol cotton ball to wipe the overnet. 2) surpluses net culture, and also experiment materials needed to put Taiwan on serum except), medium (3) under inverted microscope to observe the state of the cell, is full of culture bottle, bottle is required. 2, discard the medium and use the gun as much as possible. Add 5mlPBS liquid to the wall and shake in four directions. 3, adding 0.25% pancreatin 600 ul petri dishes, dish, up and down or so spread evenly, 37 ° digest about 30 minutes, observe, see cells sediment left, namely digested completely. 4, add in 4ml of fresh culture of 5% serum, repeatedly blow the digested cells to remove the wall and disperse it, make a cell suspension, and then install it in a 15ml centrifugal tube. 1000 RPM, 3 min. 5, abandon the supernatant. Use PBS 3ml to wash cells, blow or swirl, wash twice, centrifugal 1000rpm, 3min. For each cell, there are a total of 3 ml of diluent for each cell, with a total of 6 samples, so 20ml of diluent (no blood, 20ml + 10% BSA 200ul). Cell deposits add 3ml cell diluents, blow and mix, and dilute the cell suspension. The cell count plate is 15ul per hole. Each sample counts 2 times, the mean. (8 large total / 8 = numeric * 104) Based on the density of the pavement, the amount of the diluted cell suspension was calculated, and the remaining volume was filled with the cell diluent. Common cell counts and planks (immunofluorescence, WB, etc.) [experimental steps] Prepare to work with 75% of the alcohol to wipe your hands while wiping the overnet with an alcohol cotton ball. 2) will be The material that is required for the culture of culture and experiment should be put into the supernet for sterilization (except for the serum, medium of culture) 3) inversion Microscopically, the state of the cell, if its