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Size Sterile
Contamination and sterilization of tissue culture laboratory
Microbial contamination is a severe problem in tissue culture research. Microbial contamination often effect tissue culture studies and results very severely. Microbes which contaminate the cultures and the laboratory environment and equipments are species of bacteria, fungus and viruses.The bacterial contaminants include Pseudomonas flourescens, Escherichia coli, Proteus sp, Micrococcusspp, Streptococcus pneumoniae, Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Corynebacterium sp and Erwinia sp. While Fungi isolates were Alterneria tenius, Aspergillus niger,Aspergillus fumigatus, Cladosporium sp, Saccharomyces sp, Fusarium oxysporum, Rizopus nigricansand Fusarium culmorum. The rate of occurrence of S. aureus, B. cereus, B. subtilis and E. coli werefound to be higher (ranging from 36-46%) in human skin than in all other sampled materials. The laboratory walls and tables also harbored most of the contaminating microbes. The laboratory indoor air was found associated with all the contaminating microbes.
The technique of in vitro cultivation of plant cells or organs is primarily devoted to solve two basic problems; first to keep the plant cells and organs free from microbes,i.e bacteria and fungi, and second, to ensure the desired development in the cells and organs by providing suitable nutrient media and other environmental conditions.
All the materials e.g.,vessels, instruments, medium, plant material etc. used in culture work must be freed from microbes. Basically the technique consists of taking a piece of a plant (such as a stem tip, node, meristem, embryo, or even a seed) and placing it in a sterile, (usually
gel-based) nutrient medium where it multiplies. The micro propagation has also been used extensively in the improvement of selections of plant with enhanced stress or pest resistance, production of pathogen free plants and somatic hybridizations (Daniel, 1998).
The nutrient media in which the plant tissue is cultivated is a good source of nutrient for microbial growth.These microbes compete adversely with plant tissue culture for nutrient. The presence of these microbes in plant cultures usually results in increased culture mortality, the presence of latent infections can also result in variable growth, tissue necrosis, reduced shoot proliferation and reduced rooting (Kane, 2003). Although, the tissue culture techniques usually involves growing stock plants in ways that will minimize infection, treating the plant material with disinfecting chemicals to kill superficial microbes and the sterilizing the tool used for dissection, the vessels and media in which cultures are grown (George, 1993). However, contamination hasbeen reported as constant problem, which can compromise development of all in vitro techniques (Enjalric et al.,1988).About thirty-one micro-organisms have been reported by Leggatt et al., 1994 in tissue culture lab.The rate of occurrence of bacteria isolates was higher than that offungal isolates in the plant tissue cultures (Odutayo et al, 2007)
Laboratory walls and tables and laboratory indoor air also harbored most of the contaminating microbes. Microbes are living, biological contaminants that can be transmitted by infected people, animals and indoor air, and they can also travel through the air and get inside homes and buildings.
Tissue culture work must be freed from microbes. All culture vessels (glasswares and teflon plasticwares) are washed with detergent and then first rinsed with tap water and then finally with distilled water. Sometimes, the used glasswares are also autoclaved before washing to remove traces of agar and to destroy infection.The washed glasswares are then sterilized by dry heat in an oven at 160-180.c for 3 hours. After pouring, the culture vessels are plugged with non absorbent cotton and finally autoclaved at a pressure 1.06 kg/cm2 for 20 minutes in an autoclave. Some types of plasticwares and instruments like pipette are also autoclavable Therefore, all the culture vessels, instruments, medium and plant material etc. used in.Instruments like forceps,scalpels, needles, spatula etc. are first dipped in 95% alcohol and then sterilized over flame and then leave for dry.
Some growth regulators like GA3,Zeatin,urea, ABA, certain vitamins are heat liable hence, filter sterilized by passing through a membrane filter of 0.45u or lower pore size.
The plate form of the laminar flow cabinet, hands of the operator etc. are sterilized by wiping them with 70% alcohol.
Plant material is surface sterilized by sodium or calcium hypochlorite solution (0.3-0.6%) for 15-30 minutes. Other compounds used for surface sterilization are mercuric chloride (0.1-1%), silver nitrate (1%), bromine water (1-2%), H2O2 (10-12%) etc. but it is mainly depend on the source and type of the plant material or explant.
All tissue culture operations like sterilization and inoculation of explant,transfer of plant material are to be done in a Laminar Air Flow Bench (HM-104, horizontal type). Laminar Flow Bench is presterilized by ultraviolet radiation for 40-45 minutes. The Laminar Air Flow Bench is properly swab with rectified spirit before starting the inoculation work. Tissue culture rooms or lab should be keep clean and disinfectant.
The culture maintenance and tissue transfer rooms may be fumigated with formalin at regular intervals against bacterial and fungal contaminations. It would be advisable for the workers to wear a lab coat and acid resistant gloves. Keep footwear outside the lab, transfer room and growth chamber is also recommended. All doors and windows should be closed and all electric circuits should be of before the process of fumigation. Person should cover himself properly. Fumigation also kills insects and pests which are hiding under tables and in walls. It is an easy procedure of fumigation, in this just dip cotton in formalin and burn it and keep in 2-3 places of lab and come out, shut the door immediately. It is advisable to fumigate the lab on week ends so that it effects properly. Come early on monday, cover yourself properly open the lab doors and windows and come out. Ventilate the lab for 3-4 hours.
Generally used fumigants are- Formaldehyde, 1,3-dichloropropene, phosophine, lodoform, sulfuryl floride etc.
About the Author
i am a biotechnologist / tissue culturist.I like read and write about science. Recently i have started to write on a blog www.plantissueculture.com to share my knowledge. Beside this i have agreat interest in history.
Am I sterile and will my penis grow.?
When I jack off not that much sperm comes out as those in the videos I have seen I mean like a lot less. I stared ejaculating about a year ago. Will it progress or stay like this? Also my sperm isn't white but clear not that sticky what is it? My penis is like 3 inches long will it grow? I am 14 almost 15 and all my friends have full grown penis but me. What should I do. I have little pubic hair aswell. I am kinda fat 80 kilos does that affect my penis size?
also how are u apposed to clean up after masturbating and what happens if u didnt do that before can that affect growth?
what is pre-cum will it develop into sperm or do i have to keep on masturbating because now i still have an orgasm after i get pre-cum
You are not sterile.Just young:the guys in the videos are picked because of their performance.
And they are huge too:they have thousands who apply for a job in the sex industry,but they all have auditions too.
You fail?Camera shy?
Next!!!!!!
So do not compare yourself to them.
You are not yet making sperm:just precum.
If you loose weight your penis will look bigger.
And you are just13:all will develop in it's own pace.
That your friends are fully grown is because of their hormones.
Your hormones will be active soon too
Sky.
Having blood bath with size 7 sterile gloves