Aseptic Techniques.

Lab aseptic techniques

1. Aseptic Techniques. (Plant tissue culture )

2. What is Aseptic Techniques?? Aseptic technique refers to a procedure that is performed under sterile conditions This includes medical and laboratory techniques which deals with cultures and human cells and tissue transplantation Aseptic is a method used to prevent contamination with microorganisms

3. What is the aim of Aseptic Technique??? To prevent the access of micro-organism during the preparation and testing

4. What is a culture?? A microbiological culture, or microbial culture, is a method of multiplying microbial organisms by letting them reproduce in predetermined culture medium under controlled laboratory conditions. A microbiological culture, or microbial culture, is a method of multiplying microbial organisms by letting them reproduce in predetermined culture medium under controlled laboratory conditions.

5. Plant tissue culture is rich in source of nutrients Microbes can grow very fast on it as compared to plants, which is very harmful for plants Therefore aseptic conditions should be maintained in the culture bottle

6. Sources of Contamination Sources of Contamination 1. The Atmosphere. 2. The Breath. 3. The Hands

7. 4. The Clothing 5. The Working Surface 6. The Equipment

8. What need to be sterilized in the process of aseptic technique?? Culture Media Fluids used in the lab Reagents Laboratory containers Laboratory Equipment's

9. Methods of Sterilization A :- Physical Methods Dry heat: Hot air oven Glassware Flaming Inoculation loop and needle Moist-heat (steam under high pressure) Autoclave Culture Media

10. Filtration Radiation Used for heat sensitive liquids like, Antibiotic solutions and Sera Used for enclosed area such as inoculation room and virus lab

11. B :- Chemical Methods Alcohols Aldehydes Phenolics Here the surface, skin etc. is sterile eg : ethyl alcohol 70% conc. Here equipment such as Centrifuge is sterile eg : Formaldehyde Here floors, walls, slabs are sterile Eg : Phenols (Chloroxylenol)

12. Instruments for Plant Tissue Culture These are Sterilized by dipping in 95% ethanol followed by flaming and cooling needle Forceps blunt and sharp scalpel rod

13. Environment of the tissue culture laboratory Media room Maintain cleanliness, removal of contaminated culture, and restricted entry is done Washing room Contaminated cultured should be autoclaved and discarded with utmost care and maintain general cleanliness Transfer area A sterile area is required for performing various aseptic manipulations during tissue culture. This ensures that contaminants do not gain entry into the culture via Laminar air flow cabinets of various shapes and sizes are available commercially. These cabinets allows the tissue culturists to work in the sterile environment for long stretch of time. It provides a covered enclosed area for working

14. Laminar air flow cabinet

15. Clean hand and forearm properly before starting the work Hands may be cleaned with dilute solution of alcohol or commercial hand sanitizers available in the market Wearing a surgical mask and head gear and lab coat while working in the laminar will reduce the chances of contamination Presence of other persons in or near the transfer area should be minimized . Operator precautions:-

16. >>> Thankuh !!