Animal cell and tissue culture. Lab 1

Transcription

1 Animal cell and tissue culture Lab 1

2 Tissue culture Laboratory Safety Outline Lab Safety Biohazards Biosafety Levels Biosafety Cabinets Decontamination Biological Waste

3 Introduction A cell culture laboratory has a number of specific hazards associated with handling and manipulating human or animal cells and tissues, as well as toxic, corrosive, or mutagenic solvents and reagents. Common hazards are accidental punctures with syringe needles or other contaminated sharps, spills and splashes onto skin and mucous membranes, ingestion through mouth pipetting, and inhalation exposures to infectious aerosols.

4 The fundamental objective of any biosafety program is to reduce or eliminate exposure of laboratory workers and the outside environment to potentially harmful biological agents. The most important element of safety in a cell culture laboratory is the strict adherence to standard microbiological practices and techniques.

5 Lab Safety Safe Laboratory Practices Lab coats Safety glasses Proper footwear Hair back No food or drink in the laboratory

6 Biohazards An agent of biological origin that can cause disease in humans Microorganism Toxin Allergen

7 Biosafety The combined use of laboratory practices, laboratory facilities and safety equipment Why use biosafety practices? To protect: Workers/Students Products/Experimental results Environment/Laboratory classroom

8 Biosafety Levels BSL1 - BSL-1 is the basic level of protection common to most research and clinical laboratories, and is appropriate for agents that are not known to cause disease in normal, healthy humans.

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10 Biosafety Level 2 BSL2 - agents associated with human disease.

11 Biosafety Level 3 BSL3 - Agents with potential for respiratory transmission, may cause serious and potentially lethal infection

12 Biosafety Level 3

13 Biosafety Level 4 BSL4 Dangerous and exotic agents that poses a high risk of lifethreatening disease. Agents (all viruses) include Marburg virus, Ebola virus

14 Biosafety Level 4

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16 Biosafety Level 2 (BL-2) Use BL-2 practices when working with: Agents of moderate potential hazard to personnel and the environment. Examples of BL-2 agents: Human blood or body fluids E. coli 0157:H7 Retroviral vectors Human cells in cell culture

17 Biosafety Level 2 (BL-2) Adequate illumination Eyewash facility Negative air pressure Autoclave available Biological safety cabinet Lab must be separated from public areas

18 Biosafety Level 2 (BL-2) Standard Work Practices Use mechanical pipetting devices Wash hands frequently Decontaminate work surfaces daily Handle wastes properly Maintain insect and rodent control program

19 Biosafety Level 2 (BL-2) Special Practices Place used slides and coverslips in sharps containers, never in any other receptacle. Sharps containers are: Red in color Marked with the biohazard symbol Puncture resistant Leak proof

20 Biosafety Level 2 (BL-2) Personal Protective Equipment (PPE) Lab coat or apron Safety glasses or goggles Gloves Biosafety cabinet Aerosols Large volumes High concentrations

21 Biosafety Cabinets (BSCs) Provide product, personal and environmental protection. Various classes of BSCs are available (classi, II, III) Amount of air exhausted Amount of employee protection BSC: Class II is the most used in tissue culture.

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23 Biosafety Cabinets /Tissue culture hood Air flows straight down from the top of the hood. Sterile air is exhausted from BSCs through a HEPA filter. HEPA filter can trap particles to 0.3 microns Chemical vapors pass through HEPA filters Air flow could be horizontal or vertical.

24 High-Efficiency Particulate Air or HEPA is a type of air filter

25 Vertical laminar hood

26 Decontamination Sterilization: destroy all microbial life, including spores Disinfection: destroy a majority of microbial life, but not necessarily spores Antiseptic: germistat used on skin to inhibit growth of microorganisms

27 Decontamination Methods Heat: steam heat, dry heat, incineration Chemical: bleach, ethanol, hydrogen peroxide, ethylene oxide, paraformaldehyde Radiation (UV)

28 Biological Waste Types Cultures Sharps Pipettes, tips and weigh boats Other materials

29 Biological Waste Solids Collect and package in clear autoclave bags Autoclave to sterilize Dispose of in regular trash dumpster (as long as no chemical or radioactive contaminants are present) Liquids Collect in containers with lids Autoclave or treat with 10% bleach to sterilize Dispose of down the drain (as long as no chemical or radioactive contaminants are present)

30 Biological Waste Sharps (needles, syringes, scalpel blades, slides, blood vials, pasteur pipettes) Collect in approved sharps container Autoclave to sterilize Dispose of in medical waste boxes

31 References bco-cell-culture-basics/cell-culture-laboratory-safety.html biosfty/bmbl5/bmbl5toc.htm. =hss0&oq= &ie=utf- 8&rlz=1T4RNRN_enSA425SA428&q=biosafety+levels+labo ratory+ppt.&gs_upl=0l0l2l195403lllllllllll0&aqi=s bco-cell-culture-basics/cell-culture-equipment.html

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