Lab 3. Incubator & Biological Safety Cabinet PDF

Lab. Instrument Lab 3. Incubator & Biological Safety Cabinet Lecturer : Mariwan fathalla [email protected] 2024 1 Outline What is Incubator ? Components/Parts of Incubator...

Lab. Instrument Lab 3. Incubator & Biological Safety Cabinet Lecturer : Mariwan fathalla [email protected] 2024 1 Outline What is Incubator ? Components/Parts of Incubator Types of Laboratory Incubators Uses of Incubator What is the Biological Safety Cabinet? Classification of biological cabinets 2  Introduction Incubators Incubators are insulated boxes with an adjustable heater, typically going up to 60 to 100 °C, used in medical and other laboratories, to grow and maintain bacteria and fungi or cell cultures at temperature 37co for 18-24 hours. An incubator can be used for the cultivation of both unicellular and multicellular organisms. 3 The incubator provides viable growth factors such as : 1. Constant temperature 2. humidity is typically >80% to prevent evaporation 3. carbon dioxide (CO2) level of 5% to maintain a slightly acidic pH 4. oxygen to maintain life. 4 Components/Parts of Incubator 5 Main Parts of Laboratory Incubators 1. Air circulating fan. 2. Thermostat. 3. Timer. 4. Thermometer. 5. Electric heater 6. Some units have inner glass doors that permit the contents to be viewed without disrupting the atmosphere of the incubator. 6  General types of incubators There are many different types of incubators, and the most commonly used are the following: 1. Standard incubators. These incubators can be gravity or fan assisted convection and their range of temperature is from ambient to a maximum of 80°C or 100°C 2. Cooled incubators: These incubators work at temperatures close to or below ambient temperature 3. Humidity incubators: These incubators control both, temperature and humidity, refrigeration system. The humidity is usually obtained from a tray of water which is constantly evaporating. 4. CO2 (carbon dioxide) incubators: These incubators are widely used in maintenance and growth of Biological samples when it is necessary to maintain a constant pressure with CO2 concentration in air ranging from 5 – 20%. 5. Shaking incubators: These incubators shakes in a temperature controlled atmosphere. 7  Uses of Laboratory incubators A. biology applications, such as 1. cell and tissue cultures, cultures that involve the extraction of fragments of animal or vegetable tissue and storing them in controlled environments to analyze their growth 2. molecular biology. 3. microbiology 4. biochemical and hematological studies. B. pharmaceutical and hematological studies. C. food analysis. 8 Biological Safety Cabinets 9  Biological Safety Cabinets A Biological Safety Cabinet is a ventilated enclosure offering protection to the user, the product, and the environment from aerosols arising from the handling of potentially hazardous micro-organisms. The continuous airflow is discharged to the atmosphere via high-efficiency particulate absorber filter (HEPA) and high-energy ultra-violet light units to kill off the live bacteria and viruses trapped by the filter media 10  Uses of Protection cabinet 1. Personal Protection from harmful agents within the cabinet. 2. Product Protection to avoid contamination of the samples. 3. Environmental Protection from contaminants contained within the cabinet. 11 Classification of biological cabinet 12  Class 1 Biological Safety Cabinets High-efficiency particulate air (HEPA) filtered air The Class I BSC A) front opening. B) Sash. C) exhaust HEPA filter. D) exhaust plenum. 13  Class 2 Biological Safety Cabinets High-efficiency particulate air (HEPA) filtered air The Class II (A) front opening. (B) Sash. (C) exhaust HEPA filter. (D) supply HEPA filter. (E) Common plenum. (F) blower 14  Class 3 Biological Safety Cabinets High-efficiency particulate air (HEPA) filtered air The Class II (A) front opening. (B) Sash. (C) supply HEPA filter. (D) blower 15  Procedures for the Proper Use of Biological Safety Cabinet If users do not properly understand or operate cabinets, they will not maintain an adequate protective barrier between the operator and the experiment. Listed below are procedures for the proper use of BSCs. 1. Turn the cabinet fan and the fluorescent light on. Turn off the UV light. Confirm that the drain valve is closed. 2. Wipe the work surface with 70% ethanol or other appropriate disinfectant. Let the unit run for 5 -10 minutes to clean itself before beginning work. 3. Plan the work operation in advance. Place everything needed for the complete procedure in the cabinet before starting. Nothing should pass through the air barrier, either in or out, until the procedure is complete. Arrange materials in a logical manner such that clean and contaminated materials are segregated. Remove from the cabinet all materials or equipment not necessary for the particular procedure. 16  Procedures for the Proper Use of Biological Safety Cabinet 4. Avoid placing materials on the air intake grille, at the front of the cabinet as this disrupts the protective air barrier. 5. Keep equipment at least four inches (10 cm) inside the cabinet work area. Perform manipulations of hazardous materials as far back in the work area as possible. 6. After the procedure is completed, decontaminate all equipment in direct contact with the research agent with an appropriate disinfectant. Run the cabinet at least three minutes with no activity to purge airborne contaminants from the work area. 17  Procedures for the Proper Use of Biological Safety Cabinet 7. After removal of all materials and equipment, wipe the work surface with 70% ethanol or other appropriate disinfectant. Clean any spilled culture media that may support fungal growth and cause contamination in subsequent experiments. 8. Turn off the cabinet fan. Some researchers prefer to let the cabinet run continuously. You may turn on the UV light if the BSC is so equipped and there are barriers in place to prevent inadvertent UV exposure to other lab personnel. 18  Procedures for the Proper Use of Biological Safety Cabinet 9. Limit access to BSC to one person at a time. Two people simultaneously moving and working in a BSC can disrupt the protective air curtain at the face of the BSC. This can compromise the safety of the individuals working in the BSC, and others in the room as well as product protection, thus negating the engineering control. 10. Do not use an open flame in the cabinet as it disrupts airflow and could damage the HEPA filter. 11. Conduct research in a „clean to dirty‟ (left to right) work pattern. 19  Biohazard Spill Control Inside a BSC 1. Keep the BSC on. 2. Put on protective gloves. 3. Spray wipe walls, work surfaces, and equipment with decontamination solution. 4. Drain pans, and catch basins with decontamination solution. 5. Allow to stand for 20 minutes. 6. Drain excess solution into the cabinet base. 7. Lift out tray and any removable exhaust grille work. 8. Clean top and bottom surfaces with sponge/cloth soaked in decontamination solution 20 Thank you 21