Summary of the experience of cell crawling experiment

Summary of the experience of cell crawling experiment

The first is the treatment of coverslips. Ordinary coverslips are cut into small squares of the size you want with a grinding wheel, first washed with washing powder, rinsed with water and dried, then soaked in acid overnight, washed with running water after removing the acid. Bake dry, autoclave in a utensil, and then bake it in the oven for about 8 hours.
The climbing piece can be in a six-well plate or a 24-well plate in a petri dish. I choose to climb the piece in a petri dish. One is to save money (the petri dish can be reused) and the other is that the petri dish has a large mouth and is more convenient to operate. Disinfection of the petri dish is the same as above, remember to remove two tweezers when disinfecting.
The specific operation is: digest the cells with trypsin, fully pipetting to make it into a single cell suspension (note: this is very important, it is related to the quality of the film in the future).
Take out the sterilized petri dish, add a small amount of culture medium (to make the glass slide and the petri dish close contact), put the slide carefully into it, and then drop the single cell suspension onto the slide drop by drop. Finally, cover the petri dish and place it in a 37 degree 5% CO2 incubator for culture. According to the cell growth condition, take out the slides in a timely manner for 24 hours or more.
The slides were washed three times in 37°C PBS for 3 to 5 seconds each time, and then fixed in 4% paraformaldehyde for 15 minutes, and then rinsed with 37°C deionized water to clean the formaldehyde. Pay attention to the gentle technique, otherwise The film drop is terrible. At the same time, pay attention to the front and back of the cover glass during the operation, otherwise it is really useless.
Put the finished slide on the filter paper to dry, and then stick it on the microscope slide with neutral gum. Note that you must wait for the neutral gum to completely dry before you can do subsequent experiments, otherwise the slide will fall off, and the finished cell slide can be stored at -20 for later use. How long can it be stored? Not sure, of course try to use it as early as possible.