If you also have a greenhouse or growth rooms, make it a lab rule that you cannot enter the tissue culture room after entering those other spaces (for the day). So, all tissue culture work is done prior to work in places with loads of fungal spores, bacteria, etc.

Have dedicated cell culture items (pipets, pens, tweezers) that don't leave that space.

No wearing of coats, hats, scarves or other outdoor outer layers in the tissue culture space as those often are laundered infrequently compared to shirts/pants etc.

Empty waste containers frequently. Check for and remove contaminated specimens often. Wipe down clean hoods before and after every usage, keep them clean and organized.

If possible, have the tissue culture area away from high traffic areas.

Really, a big factor is the people, their training, and their habits. It's easy to get complacent and cut corners.

Hope this helps!

1. Personal Hygiene & Lab Attire:

• Wear a clean lab coat, gloves, and face mask (if working near laminar flow), Disinfect gloves with 70% ethanol frequently.

2. Sterile Work Environment

• Always work in a laminar airflow hood (LAF) that has been UV-sterilized for 15–30 min before use.
• Wipe surfaces with 70% ethanol or isopropanol before and after work.

3. Sterilisation of Tools & Media

• Autoclave all media, tools (forceps, scalpels), and containers at 121°C for 20 min.

4. Handling Explants & Cultures

• Surface-sterilise plant material with: 70% ethanol (30 sec – 1 min) Sodium hypochlorite (10–20%, 5–10 min) + a few drops of Tween-20 Rinse 3–5 times with sterile distilled water.

5. Preventing Cross-Contamination

• Use separate tools for different cultures.
• Avoid touching non-sterile surfaces (e.g., bottle necks, sleeves).
• Seal plates and flasks with Parafilm to prevent airborne contaminants.

6. Regular Monitoring & Decontamination

• Check cultures daily for bacterial (slimy growth) or fungal (fuzzy spots) contamination.
• Discard contaminated cultures immediately (autoclave before disposal).
• Clean incubators and LAFs weekly with ethanol or bleach solution.

7. Airflow & Lab Layout: Keep doors and windows closed to reduce drafts.

9. Proper Waste Disposal

Hey, First of all thanks a lot, can I send to you in privacy

I used

PPM

https://plantcelltechnology.com/products/plant-preservative-mixture-ppm

when I was doing pearl millet and hybrid bermudagrass tissue culture/genetic engineering. Definitely reduced/eliminated contamination. Read my publications to see how I used it.

Jason