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PRESENTATION PREPARATION AND QUIZ REVIEWERS PREPARATION FOR FINAL DAY DAILY IMPORTANT STUFF/LECTURE TO REMEMBER FOR QUIZ: DAY 1 (JUNE 10, 2024) Study Pearson chapter 1 (introduction to micro) and chapter 4 (cell structures) Study Good Laboratory Practices (GLP) ○...

PRESENTATION PREPARATION AND QUIZ REVIEWERS PREPARATION FOR FINAL DAY DAILY IMPORTANT STUFF/LECTURE TO REMEMBER FOR QUIZ: DAY 1 (JUNE 10, 2024) Study Pearson chapter 1 (introduction to micro) and chapter 4 (cell structures) Study Good Laboratory Practices (GLP) ○ FDA regulations ○ Lab Sections: The necessity of having distinct sections in the microbiology lab to prevent cross-contamination and ensure organized workflow. ○ Aseptic Technique: Detailed practices for maintaining sterility during lab procedures to avoid contamination. ○ Personal Protective Equipment (PPE): Guidelines on the appropriate use of PPE, highlighting the do's and don'ts inside the lab to ensure safety and compliance with regulatory standards. ○ Maintenance of Microbiological Cultures: Best practices for preserving the integrity of microbiological cultures over time. ○ Quality Assurance: Strategies for managing the laboratory effectively to maintain high-quality standards, including routine checks and documentation. 1 PRESENTATION PREPARATION AND QUIZ REVIEWERS PREPARATION FOR FINAL DAY DAY 3 (JUNE 13, 2024) For incubation of samples. It is used to incubate samples in MICROBIOLOGICAL TOOLS water at a constant temperature 1. Autoclave over a long period of time. ➔ Sterilization machine for media and for other tools that we use. ◦ ➔ Settings: 121 C, 15 mins, 1.5 psi ➔ Used for decontamination as well. ➔ Decontam Settings: 30 mins (prolonged) ➔ Uses steam under pressure to kill bacteria. 4. Biosafety Cabinet (BSC) ➔ Sterilization ★ Prevent contamination to the ◆ Total destruction of all samples from outside sources but organisms in the certain it does not prevent contamination object. from the analyst. ➔ Sanitizing ○ So if you’re the analyst, ◆ Surface destruction you are the only one that can introduce contamination. ○ Better practice aseptic technique. ★ BSC protects the analyst from the hazardous properties of your samples. ★ Has HEPA filter, which prevents 2. Incubator contamination. Used to incubate samples at a ★ It also has a motor that sucks in period of time and at a certain air. temperature. ★ Has negative air flow. This It maintains/regulates means, there are no air inside the temperature. BSC. Yung mga machine na may ○ Air grill sucks the air. temperature na settings, usually, ★ We don’t put alcohol lamps in for sila yung may mga calibration. prolonged period because it ○ Yearly calibration disrupts the air flow inside the BSC. 3. Water Bath Same as an incubator, but can be used to to maintain the temperature of your agar media to prevent solidifying. 2 PRESENTATION PREPARATION AND QUIZ REVIEWERS PREPARATION FOR FINAL DAY 5. Refrigerator reagents. Storage of prepared media and Used to suspend cells. (For reagents. bacterial suspensions) There are multiple types of refrigerators. 9. Top Loading Balance ★ Used to weigh solid materials like powder medium and samples. ★ Used when precision of 0.1 g. 6. Inoculating Loop ○ 2 decimal places only ❖ Used to take and transfer a small sample (inoculum) of a microorganism culture ❖ Uses: pick up colonies, purification, streaking, transferring, isolate colonies on an agar. 10. Densitometer Designed for measurement of cell suspension’s turbidity. Optical density. Estimation based on turbidity. McFarland unit. ○ Pag nakuha mo yung 7. Bacterial Incinerator same na itsura nung ➔ Uses heat produced by infrared turbidity ng sample mo rays for sterilizing waste materials based on McFarland generated during laboratory standard, it means that the procedures. Electricity to heat up. estimated total bacterial ➔ Helpful for sterilizing metals like count is nasa ganitong inoculating needles and loops. range. ➔ Flaming hot Place the tube with the broth. Automatic gives the units. ○ 1, 2, 3, 4, 5, 0.5, 0.1 The higher you go (units), the higher the CFU ○ This is estimated only. 8. Touch Vortex Mixer Vortex is an important equipment in rapid mixing of samples and 3 PRESENTATION PREPARATION AND QUIZ REVIEWERS PREPARATION FOR FINAL DAY maintaining low temperatures during the centrifugation process. ➔ Spin the samples loading in a rotor at high speed. ◆ 25 C - 2 C ◆ 10,000 RPM ➔ Golden Rule: pag umiikot, dapat balanced. 11. Heating Block Maintain precise and uniform temperatures for various processes and assays. Incubation at high temps if necessary and has precise temperature control. For DNA extraction ○ 55 C For small samples only 13. Mini Microcentrifuge (spin down) Spin down only of suspensions 12. Refrigerated Centrifuge Not used for separation of pellets (separate) to suspensions. ➔ Used to separate and isolate 6,000 - 7,000 RPM substances based on their Microcentrifuge tube density by subjecting them to high-speed rotational forces. ◆ Used to separate particles suspended in a liquid according to particle size, density, viscosity, and rotor speed. ◆ The centrifugal force leads 14. Plate Centrifuge (spin down, the heavier or denser mixing) substances to move away ❖ RT-PCR uses from the central axis. well-plates/microplates. ➔ Kaya nagkakaroon ng pellet. ❖ Makes the safe and quick spin ◆ Pellet - mass of cells; down of droplets and sediment portion that condensation very easy in accumulates during well-plates. centrifugation. ❖ Low volume reactions. ➔ It is similar to a regular centrifuge, ❖ We put optical film in the but with the additional capability of well-plates to seal it. 4 PRESENTATION PREPARATION AND QUIZ REVIEWERS PREPARATION FOR FINAL DAY 17. Laboratory Bag Mixer ➔ Homogenizer. ➔ These are blenders for solid sample preparation. ➔ Paddle blender para mas mahiwalay yung sample and mas homogenize. ➔ Sample + broth (pre-enrichment 15. Automatic Dispenser media) ★ Dispenses a controlled amount of solution. ★ Adjustable. ★ Para mas mabilis mag dispense ng volume. 18. Ultra-Low Freezer Essential for reference cultures. For housing/storage -80°C Cryogenic Storage Vials 16. Colony Counter ○ Are designed for storing To count bacterial or other biological material. microorganism colonies on agar ○ For preserving the sample plates. Another way of preserving: placing Manual counter mineral oil on the top of agar slant Pwedeng isolated, pwedeng overlapping colonies. 5 PRESENTATION PREPARATION AND QUIZ REVIEWERS PREPARATION FOR FINAL DAY 19. Water Distilling Apparatus Water purification system. Type 1 - pure water; no minerals Type 2 - distilled water Both water types are used for media prep, preferable type 1. 20. Microplate Spectrophotometer ➔ measures the amount of light that a sample absorbs ➔ Determines DNA concentration based on light absorption. 21. RT-PCR (real-time) Detects targeted genetic materials in any pathogen Can quantify, amplification Uses probes used to synthesize double-stranded complementary DNA from mRNA that can be isolated from either blood or tissue samples. 6