The Tongji lab is separated into two sections, one for conducting experiments and the other for the day-to-day life of students and professors. It is important to not contaminant the living area with gloves or hands that touched fruit flies, liquids, items in the experimental area, etc. The living area is for students and professors to relax and eat, therefore required to maintain the cleanliness. Water and snacks brought to the lab are required to be placed in the living area.
For experiments that uses grown fruit flies, gloves are not a manditory requirment because we are not going to use poisonious solution, therefore simpely washing your hands is enough, however, it is best adviced to wear gloves. On the other hand, experiments that uses larva have to come in contact with solutions like PBS which may cause irritation to eye, skin and respiratory system, which means it is manditory to wear gloves.
There are many kinds of Drosophila culture media which usually contain yeast powder. However, the live yeast medium is easy to rot, deteriorate, and emit odors, or the presence of gas at the bottom of the medium raises the entire culture medium bed, affecting the development space of Drosophila. The laboratory should select the appropriate medium according to the specific needs of Drosophila research. When preparing the medium, it should be constantly stirred to make the composition of the medium uniform, and at the same time avoid burning the medium due to high local temperature. The medium should be heated to boiling and simmer for 20 minutes. When it is cooled to around 50 oC, it is divided into high-sterilized bottles or tubes. After the medium is cooled and solidified, and there are no visible water droplets on the inner wall of the Drosophila tube, plug the bottle mouth with a high-temperature sterilized bottle stopper. The unused media should be stored temporarily in the oC refrigerator.
(2) Issues to pay attention to in Drosophila cultivationAseptic treatment: Using a high-pressure steam sterilization pot, the culture bottle, bottle stopper, and micropipette gun head used in the experiment were sterilized. Before the operation, the test bench should be sterilized with 75% alcohol, and the flyboard should be disinfected with 75% alcohol before replacing the different mediums.
Appropriate bottle stopper: The bottle stopper used in the culture tube should have good breathability. If the air permeability is poor, the relative humidity in the bottle will cause inappropriate environmental factors such as mold, and insufficient oxygen for the growth and development of Drosophila after a period of cultivation. Suitable bottle stopper is made of cotton for the inner core, wrapped in gauze, and the thin thread is tied to make a cork that is slightly larger than the size of the bottle mouth of the culture tube.
Good anesthesia effect: The specific operation methods are as follows. First, put the Drosophila culture bottle horizontally, insert the gap between the cork and the culture bottle with the needle head of the injection needle tube filled with carbon dioxide gas, and slowly push the piston to inject the gas into the culture bottle until all the Drosophilas in the bottle are anesthetized. This method can effectively avoid the operation error caused by the use of traditional ether anesthesia and the discomfort caused by ether. It can also prevent fruit flies from flying out to the greatest extent and ensure the quality, safety, and cleanliness of the experimental materials. For ease of use, the needle can also be replaced with a carbon dioxide tank, connecting the hose and the gun head to reduce the gap between the cork and the culture tube wall to avoid Drosophilas that are not completely anesthetized from flying out of the gap. When using the gun head, you should also pay attention to controlling the airflow speed to prevent Drosophilas from being washed out by the airflow during anesthesia, causing material loss.
(3) Instantly deal with the culture medium problemThe medium is the basis for the growth and development of Drosophila in the laboratory. culture medium should be replaced immediately if its color turns red, black, etc. When mold grows on the culture medium, or mold grows on the bottle stopper, the culture medium, bottle plug and the Drosophila should all be replaced. The infected Drosophila can be poured into a bottle plugged with 75% alcohol-soaked cotton to kill the mold spores.
Ensure all lab members are familiar with the lab's safety equipment. Make sure everyone knows where the nearest fire pull station, extinguisher, spill kit, first aid kit, and AED are. Know how to get out of your building and where to go after and evacuation. Organize or attend an annual evacuation drill.
Avoid eating, drinking or smoking and maintain hand hygiene when conducting biological experiments in the laboratory. Wear appropriate personal protective equipment (PPE) such as lab coats, gloves, goggles and masks when handling biological samples or pathogens. Avoid cross-contamination when using biological samples in the laboratory and handle and clean laboratory equipment and work areas properly.
Follow proper practices and laboratory safety guidelines when performing biological experiments in the laboratory. Biological samples and biohazardous substances are cleaned and disposed of on a regular basis, following proper handling procedures and container labeling requirements. Ensure proper storage and labeling to avoid leakage and misuse when using biohazardous substances in the laboratory. Assess potential risks and safety measures before conducting biological experiments and ensure that adequate safety measures are in place to deal with possible emergencies. Regularly check the safety of laboratory equipment and facilities, e.g., biological safety cabinets, automated instruments, etc. Follow laboratory rules and regulations and safe operating procedures, e.g., proper labeling and storage of biological samples, proper disposal of waste, etc. When working with biological samples in the laboratory, avoid direct contact and use appropriate tools and containers for handling.
When using a biological safety cabinet in the laboratory, ensure that it is operated and maintained correctly and that the operating procedures and biosafety levels are followed, and ensure proper ventilation and exhaust systems are in operation to minimize the build-up of harmful gases. In the event of a biosafety cabinet failure, immediately cease laboratory operations and notify the laboratory supervisor or biosafety specialist.
Conduct regular laboratory safety inspections and drills to ensure laboratory safety and emergency response capability. Ensure that adequate access and emergency exits are available and maintained in the laboratory. If a fire occurs, immediately set off the fire alarm and extinguish the fire with a fire extinguisher or emergency sprinkler system. If the fire cannot be controlled, immediately evacuate the laboratory and call the police. If a chemical spill occurs, immediately stop the source of the spill and follow the laboratory chemical safety procedures, e.g., use appropriate disinfectant, isolate the spill area