BIOD171 Microbiology Lab Notebook (1)

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Portage Learning BIO 171- Microbiology Lab Notebook Lab Notebook Bookmarks (click to navigate): Lab 1 Notebook Lab 2 Notebook Lab 3 Notebook Lab 4 Notebook Lab 5 Notebook Lab 6 Notebook Lab 7 Notebook Lab 8 Notebook Lab 9 Notebook

Lab 1 Notebook Back to Home Page Title: JG01 Introduction to Medical Microbiology Objective: To learn lab safety, how to maintain a lab notebook as well as identification and function of general lab equipment. Cultivation of samples (growth conditions) - equipment used Identification of samples (biochemical assays) - tests available Evaluation of samples (microscopy) - visualization and recognition of key characteristics Procedure: Basic Equipment Cleaning ● Autoclave ○ Heats up to 125 ℃ ○ Uses heat, pressure and steam to sterilize the environment. ○ Steam is more efficient to sterilize an environment compared to dry air. Growing ● Fixed incubator ○ Maintained around 37 ℃ ○ Able to hold petri dishes as well as tubes for liquid cultures. ○ Door is air tight and also can be covered by a metal door to control the light conditions throughout the experiment ● Shaker incubator ○ Maintained around 37 ℃ ○ Utilized when using a growth media, a liquid nutrient broth, to inoculate bacteria. ○ Shakes the culture in a swirling fashion to help aerate the bacteria,which in turn helps it to grow

Visualizing ● Microscopy Storing ● Refrigerator ○ Maintained around 4 ℃ ○ This temperature slows the bacteria's growth to preserve the samples that are being tested ○ Used when a study cannot be completed in one day Lab Safety ● Never eat or drink in the lab ○ contamination risks ● Use Personal Protective Equipment (PPE) ○ latex or thermal gloves (application dependent) ○ eyewear protection ○ lab coat ● Never leave the lab while wearing PPE ○ public hallway, bathroom, cafeteria Lab Notebook Title: First Name, Last Name Initial Exp# Title of Experiment Objective: Display purpose of the experiment Procedure: Each step of a given protocol is recorded ● Temperature/chemical used/growth conditions ● Clearly indicate any deviations that occur during experiment Notes: Additional information placed here. Results: A summary of final outcome of the experiment Notes: Remember to use lab notebook to take notes while watching lab videos. Results: Learned some functions of general lab equipment, general lab safety, and how to maintain a lab notebook.

Lab 2 Notebook Back to Home Page Title: JG02 Basics of Microscopy Objective: To become familiar with the basic components of a light microscope as well as how to load a sample for viewing. Procedure: 1. Review parts and components 2. Load sample slide onto microscope 3. Select magnification 4. Make necessary adjustments to optimize sample visualization Notes: Basic Parts: Standard light microscope ● Eyepiece (oculars): use fingers to grab and separate the pieces to fit the width of the eye. Must use both eyes. Will see a single circle in the viewfinder. Not two overlapping. ● Arm/neck: when moving microscope, hold the arm with one hand and use the other hand to carry the base. ● Objectives: provide the samples magnification. Can use arms to switch the magnification by rotating the lenses (will click when locked in place). The shortest has the least magnification. The magnifications are: 4x (shortest), 10x (yellow), 40x, 100x (largest). ● Revolving nosepiece ● Stage: flat surface where you will place your sample. Two basic ways: clamp holder (pinch 2 metal bars to hold the slides), stage clips (metal clips that rotate on the stage). Ensures the sample isn’t moving around. Has a stage guide which can help to control vertical access and horizontal access. ● Stage controls ● Stage clips ● Coarse/Fine focus: has outer which is coarse adjustment and inner knob which is fine focus (used to finetune). ● Iris diaphragm: controls how much light is let into the microscope which will affect the eye piece. Can be used to restrict how much light goes through to the sample. ● Light Source: can be adjusted using a knob to dim or brighten the light let into the sample you are trying to view. ● Base: make sure it is always steady to avoid vibrations into samples or imaging.

Visualization ● Types of objective: Dry vs Oil ● Intensity of light source: Too bright = saturation, too dark = low visibility ● Total magnification = (power of objective) x (power of eyepiece) ○ If a cell is 15mm in diameter, using a 40x objective and 5x eyepiece the cell will now appear to the eye 200x large or ~3000mm in diameter Results: Now able to identify the parts of a microscope, how it can be used to load and view a sample, how to use the functions, how to adjust magnifications and light, and how to calculate total magnification.

Lab 3 Notebook Back to Home Page Title: JG03 Mounting Techniques Objective: Microscopic examination of bacterial samples through various staining techniques. Identify color and shape of given samples. Procedure: Dry Mount 1. Clean slide (70% ethanol) 2. Circle area on slide for easy location of specimen (optional) 3. Apply organism to slide: - if from culture, use sterile loop to spread onto slide - if from plate, use sterile loop to pick colonies and mix with a drop of distilled water 4. Air dry at room temperature until all moisture has evaporated. Wet Mount 1. Clean slide (70% ethanol) 2. Circle area on slide with a wax/hydrophobic pen 3. Apply organism to slide: - if from culture, use sterile loop to spread onto slide - if from plate, use sterile loop to pick colonies and mix with a drop of distilled water 4. View under microscope - Wet mount is ideal for viewing the motility of an organism. Do not dry out. Gram Staining 1. Clean slide (70% ethanol) 2. Apply organism to slide: - use sterile loop to spread ~1-3 drops onto slide - spread into a thin film 3. Allow to air dry 4. Fix organism to slide by passing 3 times through a flame. Do not overheat slide. 5. Flood the slide with crystal violet for 30-60 seconds

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