MCB 3020L PRACTICAL STUDY GUIDE Module 1 – Biosafety Guidelines: o Lab Space Wipe down lab bench BEFORE and AFTER performing experiments o Personal Protective Equipment Closed-toe shoes Tie back long hair Lab Coats (At all times when in the lab) Wear gloves if… Hands have fresh cuts/abrasions When staining microbes Handling hazardous chemicals o Hand washing is important: Hand transmission important route of infections Eat, Drink, Eyes, Mouth, Nose, Open Wounds Hands easily contaminate during lab procedures Hand-to-face contact common IMPORTANT: Wash hands especially after removing gloves o Biohazard Spill Notify, Cover with absorbent material, and disinfect (sit for 30 min), dispose in biological waste bin Never pick up broken/sharp objects with hands o Waste Disposal Biological Waste: Solid or liquid waste which may present a threat to animals, crops, or natural ecosystems Biomedical Waste Biological Toxins Biomedical Waste o Treat all microorganisms as pathogenic; handle unknown bacteria with caution. o Sterilize inoculating loops/spreaders by flaming; use caution with hot inoculating loops o Keep loose sleeves, hair, etc away from the Bunsen burner flame . The “Beak Method” of Protective Glove Removal: o 1. Pinch cuff and pull glove away form wrist. o 2. Scoop cuff with middle finger. o 3. Form beak, pulling glove inside out over all fingers and thumb. o 4. Pinch cuff of opposite glove with beaked hand. o 5. Discard both gloves o 6. Wash your hands.
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MCB 3020L PRACTICAL STUDY GUIDE
Module 1 – Biosafety Guidelines:
o Lab Space Wipe down lab bench BEFORE and AFTER performing experiments
o Personal Protective Equipment Closed-toe shoes Tie back long hair Lab Coats (At all times when in the lab) Wear gloves if…
Hands have fresh cuts/abrasions When staining microbes Handling hazardous chemicals
o Hand washing is important: Hand transmission important route of infections
Eat, Drink, Eyes, Mouth, Nose, Open Wounds Hands easily contaminate during lab procedures
Hand-to-face contact common IMPORTANT: Wash hands especially after removing gloves
o Biohazard Spill Notify, Cover with absorbent material, and disinfect (sit for 30 min),
dispose in biological waste bin Never pick up broken/sharp objects with hands
o Waste Disposal Biological Waste: Solid or liquid waste which may present a threat to
animals, crops, or natural ecosystems Biomedical Waste Biological Toxins Biomedical Waste
o Treat all microorganisms as pathogenic; handle unknown bacteria with caution.o Sterilize inoculating loops/spreaders by flaming; use caution with hot inoculating
loopso Keep loose sleeves, hair, etc away from the Bunsen burner flame.
The “Beak Method” of Protective Glove Removal:o 1. Pinch cuff and pull glove away form wrist.o 2. Scoop cuff with middle finger.o 3. Form beak, pulling glove inside out over all fingers and thumb.o 4. Pinch cuff of opposite glove with beaked hand.o 5. Discard both gloveso 6. Wash your hands.
Wet mount lab: o Background : You can study the ecology of the ecosystem and examine and observe
changes in population composition of microbes such as amoebas, protozoa, rotifers, insect larvae, algae, and bacteria.
o Purpose : This experiment provides you with an opportunity to experience a whole new world of microscopic organisms in a drop of water. You will find examples of many different organisms, including prokaryotes and eukaryotes, unicellular and multicellular organisms, protozoans, and algae.
o Methods : 1. Using a transfer pipette add a small drop (approximately 20 uL) of sample
to a pre-cleaned microscope slide.
2. Place the edge of the microscope cover-slip on the microscope slide so that it slightly touches the edge of the sample.
3. Lower the microscope cover slip over the sample.4. Place the microscope slide with the sample on the stage of the microscope
and secure in place with stage clips.5. Use the 10x objective first.
Terms: o Biosafety – Preventative measures to avoid unintentional exposures to or accidental
release of pathogens/toxins. Established by minimizing risk to yourself, others, and the environment through compliance with regulations for biohazards
o Condenser – the part of the microscope that controls the concentration of light shone on the specimen.
Module 2 – Gram staining lab:
o Importance: The Gram reaction is usually the first test for diagnosis of a pathogen Gram stain results can have dramatic effect on patient care
o Background: Know the difference of cell wall structure between Gram + and Gram –
organisms. Which of the organisms we use fall in which category? Understand different bacterial shapes and arrangements Endospore formation Recognize images of spore stain, negative stain, flagellar stain and acid fast
staino Procedure:
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