aseptic techniques in microbiology

This method of preventing unwanted microorganisms from gaining access is termed aseptic technique. It also ensures that any liquid culture on the loop will run down into the flame. The most important part of a laminar flow hood is a high-efficiency bacteria-retentive filter, i.e., the HEPA (high-efficiency particulate air) filter. How To Streak For Isolated Colonies (SFIC): Mixed cultures (more than one species) can be isolated using the streak plate technique. This will prevent any foreign contaminants from coming in contact with the customers and sample during testing. Start the operations only when all apparatus and materials are within immediate reach. Access to physical locations is limited; masks are required. To protect patients from harmful bacteria and other pathogens during medical procedures, healthcare providers use aseptic technique. Lift the bottle/ test tube with your left hand. This site uses Akismet to reduce spam. On opening a test tube or bottle, the neck must be immediately warmed by flaming (see below) with the vessel held as near to horizontal as possible and so that any movement of air is outwards from the vessel. Position the handle end of the wire in the light blue cone of the flame. It is a fundamental skill for working in a microbiology laboratory. You will check them next lab period for growth. It is recommended to wear gloves. This leaves the little finger free to take hold of the screw cap/ cotton wool plug of the bottle/ test tube. it will contain culture, which may splutter on rapid heating and possibly release small particles of culture, forming an aerosol. A laminar flow unit (or hood) is a sophisticated appliance that can further help prevent contamination of reagents and biological cultures. Special precautions must be taken to prevent the formation of aerosols when working with BSL2 organisms. The complete removal of all microorganisms including their spo…. Ethanol disinfection is recommended because of its rapid action. Aseptic techniques should be used for decreasing the possibility of bacterial contamination. Use the incinerator to sterilize the loop. In the microbiology lab we use aseptic technique to: Many of the microorganisms we will be working with in lab are known pathogens. Place both your colored practice tube and your sterile water tube in your left hand with the lids pointing up. Loosen the cap of the bottle so that it can be removed easily. In microbiology aseptic technique is required, and involves being constantly mindful of each action performed in the laboratory environment. Place your labeled inoculated plate upside down in the rack for incubation. Wipe the outside of the containers, flasks, plates, and dishes with 70% ethanol before placing them in the cell culture hood. The technique to reach aseptic conditions is more specific, rigorous, detailed and thus complex. The general idea is to decrease the bacterial concentration with each swipe. This consists of a circular heating element. Avoid this by squeezing the teat before placing the tip into the liquid. Prevent contamination of the specific microorganism we are working with. You will spend a lot of time in lab transferring organisms from one tube to another, or to slides or to plates. Take care! Aseptic technique and clean technique are two closely related healthcare practices that both aim to keep people safe from infection. Firmly attach the hose to the tapered gas line. © 2020 Microbe Notes. If cotton wool plugs have partly lost their shape, they can be more easily guided back into the neck of the vessel by slowly twisting the mouth of the vessel as the plug is pushed down. This common piece of equipment burns a continuous stream of a flammable gas—usually natural gas (methane)—based upon a design made almost 150 years ago by the German chemist Robert Wilhelm Bunsen (1811-1899). Close windows and doors to reduce draughts and prevent sudden movements which might disturb the air. Sterile graduated or dropping (Pasteur) pipettes are used to transfer cultures, sterile media. Depress the teat cautiously and take up an amount of fluid which is adequate for the amount required, but does not reach and wet the cotton wool plug. In the microbiology lab we use aseptic technique to: 1. Incubate at 37°C for 24 hours. Broth to Broth Transfers Using BSL2 Procedures, Broth to Plate Transfers Using BSL2 Procedures, Plate to Plate Transfers Using BSL2 Procedures, Plate to Broth Transfers Using BSL2 Procedures, aseptically transfer organisms from broth/plate cultures using, handle biohazard spills and dispose of biohazard materials. They will look like small dots on your plate. If you remove a cap or cover, and have to put it down on the work surface, place the cap with opening facing down. Truckee Meadows Community College is northern Nevada's jobs college, preparing qualified students for jobs in industries right here in Nevada. Many of the microorganisms we will be working with in lab are known pathogens. After carrying out the procedure required, for example, withdrawing culture, replace the cap/ cotton wool plug on the bottle/ test tube using your little finger. Immediately after use put the contaminated pipette into a nearby discard pot of disinfectant. . Look at the underneath of the striker. Although not all microbes are harmful to humans, some may cause infec- tion and disease. Ensure the full length of the wire receives adequate heating. HEPA filter technology was declassified after World War II, allowing extensive research and commercial use. Used correctly, it provides the work space with clean, ultrafiltered air. Flame the mouth of the tube with the Bunsen burner and replace the cap. TMCC offers over 50 programs of study that lead to more than 160 degree, certificate and other completion options. 3. You need to first re-suspend them in the broth. The bottle will be hot. Prevent contamination of the room and personnel with the microorganism we are working with. For instance, there … Before lighting a Bunsen burner, set the air and gas adjustments to a minimal open position. It requires knowing which viruses or bacteria are harmful to the product at hand, and how to remove them while keeping helpful microorganisms intact. Prevent contamination of the specific microorganism we are working with. You have now transferred your organism to a fresh tube. Touch the surface of the loop to the agar surface. Use the same plate of bacteria you did for your plate-to-plate transfer. Research and industrial laboratories working with highly sensitive Close the lid of the plate. Be careful not to talk, sing, or whistle while performing sterile procedures. Aseptic technique is essentially the backbone of microbiology. Zigzag the last part into the center of the plate. Do not stretch over the table for what you need. You want to know where you’re working from in your plate so you aren’t standing there with a cooling loop wondering what colony would work the best. Biology educators emphasize aseptic technique when teaching lab courses. Repeat by dragging back and forth. This is the coolest area of the flame. B.Sc. These two media provide everything the microorganisms need. Cleaning and disinfecting lab surfaces prior to use, limiting the duration that cultures or media are uncapped and exposed to the air, keeping petri dishes closed whenever possible, effectively sterilizing inoculating loops and other equipment that comes into contact with cultures or media, and. Keep in mind that you must wear the correct Personal Protection Equipment (PPE) in all labs where you are using microbial cultures, stains, chemicals, and glassware or microscope slides! avoiding breathing on cultures or sterile instruments. If you get burned you run cold water on burn area for 15 minutes. These techniques usually involve disinfecting working areas, decreasing the possible access by bacteria from outside air to the media and using of flames for killing bacteria that might enter the … A Bunsen burner is not practical in some situations, e.g., within a laminar flow unit where the heat will disrupt airflow. Always wipe your hands and work area with 70% ethanol. Turn off bunsen burner or hot plate when not using. Lab Report 1: Preparation Of Culture And Aseptic Technique In Microbiology In this way there is perhaps less chance of spores settling onto the plate from the air. Discard contaminated material in the appropriate container. The hot part of the flame is above the inner bright blue ‘cone’ and the vessel needs to be moved through the flame, not held in place. In this lab you will be learning standard microbiological procedures appropriate for Biosafety Level (BSL) 1 and Biosafety Level (BSL) 2 precautions. Instead of using a Bunsen burner to “flame” loops and other inoculating utensils, BSL2 procedures require the use of an incinerator. Insert the loop into the broth without touching the sides of the tube. Modify the air and gas adjustments until you get a hissing, small, blue flame within a taller lighter blue/violet flame. Put the colored practice tubes in the appropriate rack on the “Save” tray on the front bench. This is a little easier because you only have to hold one tube in your hand. Place your loop in the mouth of the incinerator briefly for 2-4 seconds to sterilize it. Open the lid of your dish with your left hand and hold it ajar. Asepsis is the state of remaining free from pathogenic and contaminating microorganisms.This technique ensures a contaminant free environment while handling micro organisms. Be careful! Grasp the lid off the tube and flame the mouth using the Bunsen burner. Antiseptic. Aseptic technique is important for wine microbiology for identifying and culturing organisms. Never uncover a sterile flask, bottle, Petri dish, etc. While vessels are open, all work must be done close to a Bunsen burner flame where air currents are drawn upwards. Both methods are presented in the form of … The flaming procedure should heat the tip of the loop gradually. If you can’t seem to get your burner lit, ask your instructor or IA if the main gas control is on. Save my name, email, and website in this browser for the next time I comment. Use only sterile glassware and other equipment. Although it can be assessed throughout the semester by examining the students’ technique and observation of contamination, this tool offers a simple semester-long graded assessment of each student’s technique. Have your two plates on your lab bench. Aseptic - “an environment or procedure that is free of contamination by pathogens”. Place the plates of E. coli on the “Save” tray. Flame the mouth of your tubes and replace the caps. A certified HEPA filter must capture a minimum of 99.97% of dust, pollen, mold, bacteria, and any airborne particles with a size of >0.3 μm at 85 liters/min. Gathering accurate data from plated cells and microorganisms requires a pure culture. TMCC provides a wealth of information and resources. Many of our former students comment that this is the most important thing they learned in lab! Remove the pipette from its container/ wrapper by the end containing a cotton wool plug, taking care to touch as little of the pipette as you need to take a firm hold. 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