Aseptic operation in the laboratory

(i) Pre-sterile operation preparationCulture
material vaccination sterile operation process in addition to the above-mentioned disinfection and sterilization operations, but also need to complete the following sterile operation steps, in order to obtain inoculated sterile culture materials.
Staff need to wash their hands and arms with soap before sterile operation, and wash them with running water, and after disinfection of the arm wearing sterilized work clothes, work hats and
masks
, change slippers, in order to enter the sterile operating area.
If you enter the strat flow operating room for experimental operation, you should complete the shower in the buffer preparation area, replace the sterilization cap, slippers, arm disinfection, secondary replacement of sterilization protective cap,
gloves
, slippers, etc. , and then in the wind shower area after the sterile wind shower can enter the strat flow operating room. After entering the sterile operating area, wipe the hands and forearms with 70% to 75% alcohol to complete the preparation of the sterile operation.
scrubbing the work surface with 70% to 75% alcohol, remove the sterilized laboratory supplies and place the operating equipment on the sterile device
stent
before starting the experimental operation. During the experimental operation, all operating instruments are sterilized after each use, often using alcohol lamp flame burning sterilization.
(ii) Sterile operation steps
After the preparation is completed, the implants from the natural growth conditions, according to the above-mentioned culture material disinfection method after sterilization, put into the sterilized Petri dish, and then placed under the ultra-clean work table alcohol lamp flame, with sterilized scissors and other equipment for proper separation, cutting or other post-treatment standby.Gently open the stop (cap) of the culture container at the alcohol lamp flame, rotate the bottle at the lamp flame and burn, place the culture material on the culture liquid with tweezers, dip the tweezers in the alcohol bottle, burn the alcohol lamp back into the bracket, and then quickly burn the plug (cap) for a few seconds before plugging back into the bottle.
The sterile operation of the successor culture material, the plug (cap) needs to be opened at the lamp flame, the successor material is a liquid cultured cell directly with sterilization
straw
The amount of material is placed on a new culture solution, after the substitute material for other
tissue
, with sterilized scissors or other equipment for proper cutting of the culture material, with sterilized tweezers placed on the new culture (medium), and then quickly burn the bottle plug (cap) for a few seconds to plug back into the mouth.
(iii) the source of pollution and its manifestations
when culture materials, transfer vessels, sterile operating environment and other disinfection and sterilization is not complete, culture materials carry
microorganisms
(microorganism), when it is transferred to nutrient-rich culture, microorganisms multiply rapidly, a variety of contaminated plaques.produce metabolic waste that is toxic to plant tissue when microorganisms grow, resulting in the death or loss of use of cultured materials. During the culture process, mucus or turbid traces of water appear near the culture material, and fermented foam, which is bacterial contamination.
In bacterial contamination, special attention should be paid to a milky white spore contamination, which can appear on the surface of culture fluid, or drop-shaped cloud-like presence in culture fluid, if the emergence should be strictly sterilized. And the surface of the culture liquid appears yellow, white, black and other different colors of mold,
fungus
pollution.