Methods of cleaning and sterilizing cell culture instruments and knowledge of biochemistry are learn

During tissue culture, ex vivo cells are sensitive to any toxic substances. Toxic substances include disintegrated microorganisms and cellular residues, as well as non-nutritive chemicals, so culture supplies such as new and reused petri dishes should be cleaned and disinfected strictly. The Creature Gang has an introduction there, I've seen it there, you can check it out.

Instruments, Materials and Reagents] 1Instruments: clean bench, drying oven, pressure cooker, filter.

2.Materials: UV lamp, microporous membrane.

3.Reagents: 75% alcohol, neo-gelfen, potassium permanganate, potassium dichromate, concentrated sulfuric acid.

Soaking: Newly purchased glassware often has dust, is weakly alkaline, or contains harmful substances such as lead and tablet, so soak it in tap water overnight and wash it with water, and then soak it with 2% 5% hydrochloric acid overnight or boil it for 30min and wash it with water. Essentials:

After cultivation, the glassware should be soaked in clean water immediately, so that the next brushing work can be carried out smoothly. After use, glassware often has a large amount of protein attached, which is not easy to brush off after drying, and should be soaked in water immediately after use. Brushing:

Use a soft-bristled brush and high-quality detergent to brush off impurities on the utensils, rinse and dry. Essentials: After soaking glassware, use a brush dipped in detergent to remove impurities on the utensils, and brush too many times, which will damage the surface finish of the utensils, and the detergent has a tendency to increase the pH, so it is easy to choose soft brushes and high-quality detergents.

It is forbidden to use decontamination powder. Because it contains sand particles. It can wreak havoc on the finish of glassware.

Pay special attention to the corners of the bottle. Rinse the detergent before acid soaking. Pickling:

Soak the utensils in the cleaning solution for 24 hours, and not less than 4 hours if used urgently. The cleaning solution is prepared from potassium dichromate, concentrated sulfuric acid and distilled water, which has a strong oxidizing effect and strong decontamination ability. After soaking in the cleaning solution, the trace impurities left by the glassware can be completely removed.

Rinse: Rinse fully with tap water first, flush with straws for 10min, fill and pour out each bottle, and repeat more than 10 times. Then rinse 3 times with distilled water, leaving no dead corners.

Leave to dry or tumble dry for later use. For the utensils that have been used, all polluted ones must be boiled for 30min or soaked in 3% hydrochloric acid overnight, and those that are not polluted do not need to be sterilized, but they still need to be brushed, the cleaning solution is soaked overnight, rinsed, etc. Precautions for the preparation and use of cleaning solution1

Cleaning solution (see Table 2-1 for formula): Ingredients: weak acid, sub-strong liquid, strong liquid, potassium dichromate (g), water (ml), concentrated sulfuric acid (ml), sulfuric acid concentration (v), 501000908%, 100100016014%, 6020080080% It is advisable to use acid-resistant plastic barrels or stainless steel barrels. First, dissolve potassium dichromate in water (stirring with a glass rod to dissolve, sometimes not completely dissolved). Slowly add concentrated sulfuric acid, do not rush too much, otherwise it will produce heat and cause danger (never pour the heavy acid bait into concentrated sulfuric acid).

The cleaning solution is brownish-red when it is prepared, and when it turns green, it indicates failure. Due to the extremely corrosive nature of cleaning fluids, care must be taken and protected when preparing and applying them.

It's too advanced to understand. Hard!

In general, cell culture rooms are used in sterile laboratories.

Formulate peracetic acid as a disinfectant. The laboratory has 84 or Jieerdi as a disinfectant.

Ultraviolet lamps are installed in the laboratory. There should also be UV in the safe. If you don't have one, you can buy the hand-held one. After the laboratory is finished, open the ultraviolet radiation for half an hour. If you feel that it is necessary, you can also fumigate with formaldehyde for a day. This one is more troublesome.

Laboratory equipment counter placement.

The pipette heads, syringes, and waste liquid used in the experiment are directly thrown into the peracetic acid in the safety cabinet and soaked for half an hour before being discarded.

The utensils that can be used in contact with the virus must be wrapped (the gun is generally not used, try not to touch the virus liquid), and the safety cabinet must be removed and extinguished.

Let's make a bigger autoclave. Otherwise, you will die bitterly.

Garbage is collected exclusively. If the logistics cannot be handled, it should be sterilized and discarded by itself.

Experiments must be done in a level 2 or higher safety cabinet. Make it with more straw paper underneath. If you have the conditions, you can pad aluminum foil or something, and that is impermeable.

The experimental room strictly controls the flow of personnel!! Those who can get a vaccine get a vaccine.

Wearing protective clothing to protect yourself from damage is also protecting others!!

Also, don't go out of the lab with gloves on!!

For details, please refer to the rules and regulations of foreign biosafety laboratories. Implemented on their own terms.

In the humble opinion below.

The method mentioned upstairs is generally correct, but if peracetic acid is used for disinfection, it will cause a large degree of corrosion of the intercellular device. At present, the more popular is the dry mist hydrogen peroxide disinfection technology.

The use of the world's advanced Orpheum oxy-30000 dry atomized hydrogen peroxide disinfection machine, derived from the EU GMP design, itself meets the requirements of clean area purification, does not bring into any external pollution of the same collapse infiltration of the high-efficiency clean room space microorganisms, to achieve the goal of aseptic purification, has been widely used in domestic biopharmaceutical enterprises, 70% of the top 100 pharmaceutical companies are using Orpheum oxy-30000 dry atomization hydrogen peroxide disinfection machine, As a result, it has a proven validation scheme with advantages including:

With Norford disinfectant, the sterilization effect is good, and it has 6 logarithmic killing rates for steatobacterium thermophilus spores, which can kill more than 200 kinds of microorganisms including mycoplasma, fungi, viruses and spores, and is colorless, non-toxic and residue-free, safe and reliable, and friendly to personnel;

Stable performance, non-electric heating principle, reducing the risk of high temperature easy damage to the equipment;

The verification information is complete, the sterilization equipment and the reagents used are all filed in China, with CANS certification test reports;

The equipment is small in size, easy to operate, and flexible to move to meet different space needs.

Cleaning all kinds of glassware used in plant tissue culture, especially culture flasks and vessels containing culture media, must be strictly cleaned to prevent oil, heavy metal ions, acids, alkalis and other harmful substances from remaining in the bottle and affecting the growth of cultures. Used glassware should be cleaned in time, first pour out dirt such as culture medium and culture, then immerse in water, and then wash. The washing of glassware can be carried out in different ways according to the degree and nature of the contamination of the utensils, usually alkaline washing and pickling.

All utensils contaminated with microorganisms must be autoclaved and boiled to kill the bacteria, otherwise spores will fly, pollute the environment, and bring serious difficulties to tissue culture. After the utensils are washed, they should be dried or dried and placed in a specified place for easy access.

Commonly used sterilization methods can be divided into two categories: physical methods such as dry heat (baking and burning), moist heat (atmospheric pressure or high-pressure cooking), radiation treatment (ultraviolet, ultrasonic, microwave), filtration, cleaning and a large amount of sterile water rinsing and other measures; The chemical method is to use mercury, formaldehyde, hydrogen peroxide, potassium permanganate, lysol, bleaching powder, sodium hypochlorite, antibiotics, alcohol chemicals. These methods and agents should be appropriately selected according to different materials and different purposes in the work, and the moist heat sterilization (culture medium) medium should be completed within 24 hours after preparation.

Autoclaving is aimed at e.g. sterile water, cultivation media, inoculation utensils, which can extend the sterilization time or increase the pressure. The medium should be strictly adhered to the holding time. Some cloth products, such as lab coats, masks, etc., can also be autoclaved.

After washing and drying, pack it in a high-pressure plastic bag and autoclave for 20-30min.

Cauterization (instruments for aseptic operation) During aseptic operation, tweezers, scissors, scalpels, etc. are immersed in 95% alcohol, and the fire bacteria are removed from the flame of the alcohol lamp before use. After cooling, use immediately.

Dry heat sterilization (glassware and heat-resistant appliances).

Filter sterilization (not heat-tolerant substances) Some growth regulators such as gibberellin, zeatin, abscisic acid, and certain vitamins are not heat-resistant and cannot be treated with autoclaving, usually using the filter sterilization method.