1. Disinfect the incubator: scrub with pure water first, then scrub with 95% alcohol, and then pull it under ultraviolet light! Pay attention to the cleanliness of the surrounding environment and it will not be so easily polluted! 2. My experience is that when replenishing water, it is best to heat the water to reach the temperature of the incubator. 3. The cells were nourished so early. We usually scrub with 70% ethanol, then scrub with 0.1% Xin Jie Er to wipe off, then use potassium permanganate plus formaldehyde 1: 1 ratio to smoke again. Note that if smoked, be sure to leave immediately after mixing, and then close the cell culture room, the taste is very irritating. It is best to sterilize before feeding cells, otherwise, there is really no place to put cells. 4. Our method is relatively simple, first wipe the inner wall with 75% alcohol, ventilate for 10 minutes, and irradiate with ultraviolet light for 30 minutes. 5. Our usual treatment is to wipe the inner wall with 75% alcohol first, then fumigate with formaldehyde + potassium permanganate for one night, and then ventilate for a whole day (while turning on the ultraviolet radiation). 6. We took out the plate in the incubator and put it under high pressure. We wiped the wall of the box with 75% alcohol, replaced it with Shiroshi, and then irradiated it with ultraviolet light overnight. The effect is very good. Other experts please advise. 7. Blowing one day after fumigation is absolutely not enough. At least one week is required to let formaldehyde dissipate, otherwise the cell growth is not good 8. Some incubators can heat the inner wall to 90 degrees, and some have UV function. Routine should be scrubbed with alcohol once a week. For specific operations, please refer to the manual of the incubator, or consult the incubator technical support. Care should be taken when using disinfectants. There are sensors in the incubator to detect the temperature, humidity and carbon dioxide concentration. Corrosive disinfectants or organic solvents may damage some devices. Please use them with caution. In addition, volatile formaldehyde is toxic, flammable and explosive. If formaldehyde remains in the incubator, the cells will die out (the true story of my laboratory). In short, look at the incubator manual before contacting technical support to be sure. 9. It is better not to put anything in the routine! The role of copper sulfate is to inhibit the growth of bacteria (but most of them are used to treat contaminated well plates). Because the current incubator has antibacterial function. 10. The practice in our laboratory: After autoclaving the plate, add an appropriate amount of sterilized double-distilled water or physiological saline to the aseptic operating table. Add a small amount of chlorpheniramine, the concentration is okay. Cell culture can be used. 11. I suggest 1 It is best to disinfect the incubator thoroughly. The specific method is to clean the incubator with 75% alcohol. 2. Strictly follow the principle of aseptic operation during operation 3 I think the problem of replacing the double antibody is not significant unless you are using contaminated antibiotics. 4 Minimize the number and time of viewing cells. 12. The method of cleaning the incubator in our laboratory: wash it with washing powder or dishwashing liquid-wash with clean water-84 disinfectant wash-wash with clean water-scrub with 75% alcohol 13. This is the routine incubator disinfection method in our laboratory (usually once a month), I hope it will be useful to you 14. First, take out all the contents in the incubator and drain off the three distilled water in the incubator; repeatedly rinse the incubator with three distilled water, then wipe it three times with 75% alcohol, and then three times with three distilled water; The shelf in the incubator was washed repeatedly with tap water, then wiped three times with distilled water, triple distilled water, and 75% alcohol, and sterilized under ultraviolet light for about 1 h; finally, the triple distilled water (with a little dye inside) was added to the culture Box, place the sterilized shelf immediately 15. The incubator should be placed in a sterile room, and an appropriate amount of sodium azide (antiseptic) or copper sulfate should be added to the water, paying attention to air disinfection. 16. There are two common methods for disinfecting incubators: Remove the cells, remember to tighten the cap, usually placed in a super clean table at room temperature for an hour or two, the problem is not big. If the cultivation room has air conditioning, it can also be turned on. Then wipe the inner wall of the incubator with a 75% alcohol cotton ball first, being careful not to miss any corner. After wiping twice, wash the inner wall with one or two distilled water, which can be washed several times. Finally, wipe the inner wall with disinfected cotton ball or 75% alcohol cotton ball again. The entire operation is not necessary under sterile conditions. Finally, add methylene blue to the water in the incubator and mix well. Methylene blue solution can play a certain role in bacteriostasis. When the alcohol in the incubator has almost evaporated, place the cells in the incubator. Another method is to clean it with distilled water directly, and then use the ultraviolet lamp to illuminate (preferably the one with the ultraviolet lamp in the incubator, it is much more convenient.) Usually the incubator needs to be cleaned regularly. Our room is cleaned once every 1-2 months. Of course, this depends on how much cells are raised and how often the incubator is used. 17. There have been similar incidents in our laboratory fluids. I will tell you our handling process and hope it will help you: First sterilize the incubator. If you have two or more incubators, it's easy to handle. If there is only one and there are many other cells, it's troublesome. Can only be temporarily placed on the ultra-clean table. The details are as follows. At noon, the temperature of the air conditioner is maximized (only 31 degrees), and the ultra-clean table is turned on by ultraviolet light. Turn off the UV in the early afternoon, close the cap of the cell culture bottle, transfer to the ultra-clean table, scrub the incubator with 75% alcohol, and then irradiate with ozone-free removable UV for half an hour, then put the cells back and loosen Just open the lid (remember to close the CO2 bottle valve tightly, otherwise the gas will run out). If the laboratory grows mold, it must be disinfected. Re-sterilize the laboratory, first remove the indoor air conditioner and disinfection cabinet filter screen, add a little potassium permanganate in the Erlenmeyer flask, put it on the laboratory floor, and then pour formaldehyde, just close the door and leave. After 2 days, I opened the door and loaded things, and started working as usual. Since you can't do things for 2 days, personal cells must be planned first, and everyone can rest for two days. 18. In our laboratory, all the cells are taken out, the culture phase is scrubbed with alcohol, and then irradiated with ultraviolet light for more than 4 hours, all the contaminated cells are discarded. There is also a UV disinfection in the cultivation room 19. I have also encountered this situation. My basal medium is contaminated. Under the light microscope, small black particles can be seen at the bottom of the bottle, but the specific reason is unknown. Approach: 1. All contaminated cells are thrown away after exposure to ultraviolet light for more than 2 hours; 2. The same batch of culture medium, used cell cleaning solution, etc. are all re-filtered and sterilized (a little reluctant to throw away all, I tried it, the result proved to be possible); 3. Open the incubator, scrub with 75% alcohol, and then scrub with Xinjieer (we repeated this step twice, the next day), and irradiated with ultraviolet light for more than 4 hours; 4. Clean the incubation room, ultraviolet light for more than 4 hours, repeat once; Later it was gone. :) If the pollution is serious, repeat it again and again. 20. I also encountered two major outbreaks years ago, first with fungi and then with bacteria. It made me lose confidence in cell culture. I have nourished cells for a year and a half. I usually have no cells. I feel that nourishing cells is a simple matter, but when the pollution erupts, I really feel unbearable. I remember that almost all the cells were lost at that time. Nearly a hundred bottles, only three or two bottles of seed preservation, cold !!! As for the treatment method, that is, lactic acid fumigation (remember to open the incubator and ultra clean table) one week Close the doors and windows and stop entering the cultivation room. After a week, 75% alcohol is thoroughly sterilized, that is, the alcohol is used to apply all planes in the incubator and the cultivation room, even the ground. It has been four months now, and the cells have been very good. It is recommended not to panic, the so-called soldiers will block, the water will cover. Ha ha ha, everything will be fine. 21. It seems that you are going to go all out. First consider the problem of culture medium. You can place a bottle of medium in other incubators for 24 hours to see if the medium is contaminated. However, it is better to throw away all reconfigurations. Second, consider the problem of incubators and incubators. Basically, the incubator should be thoroughly disinfected once a month. First scrub with 75% alcohol, the general incubator can be detached, disassemble the detachable part, scrub and put it in the oven (180 degrees for 5 hours). The remaining incubator was scrubbed with glutaraldehyde and then repeatedly irradiated with ultraviolet lamps. Then sterilize with UV light for 2 hours before using the incubator incubator. Your photos should show bacterial contamination, cough is better to deal with. From: http: // Led Make up Mirror,Cosmetic Mirror With Lights,Led Dressing Table Mirror,LED Cosmetic Mirror,desktop makeup mirror Zhongshan Seven Cool Electronics Technology Co.,Ltd , https://www.gdsevencool.com
Instructions for cleaning the cell incubator