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Essential Microbiology: Microscopy and Staining Techniques, Exams of Nursing

Key concepts and questions related to essential microbiology, focusing on microscopy and staining techniques. It provides an overview of topics such as the definition of a nanometer, the relationship between resolution and contrast, the use of microscope components to control light, the principles of different microscopy techniques (e.g., fluorescence, phase-contrast, dark field, confocal), the interpretation of electron microscopy images, and the application of gram staining and other staining methods. Likely intended as study material or exam preparation for a university-level microbiology course, covering fundamental knowledge and skills required for understanding and working with microscopic biological samples.

Typology: Exams

2023/2024

Available from 08/15/2024

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Med C
BIOD 171 Essential Microbiology Portage Learning Module
3 Exam Questions and Answers
Question 1
. A nanometer is defined as:
A. 10-3
B. 10-6
C.10-9
D. 10
-12
C. A nanometer is defined as one-billionth of a meter.
2. True or False: A nanometeris longer than a micrometer. False. A nanometer is 1,000 times
smaller than a micrometer.
1. Resolution and contrast are two critical factors that influence your ability to see an object.
Explain each. Resolution refers to the distance between two objects at which the objects still
can be seen as separate. Poor or low resolution means two (or more) objects may appear as
one.
*Contrast is the difference in light absorbance between two objects. Poor contrast gives a
high background and makes the visualization of multiple objects difficult. For instance,
trying to identify 2 dark colored objects at night (low light = low contrast) versus the same 2
objects in the middle of a sunny afternoon (bright light against 2 dark objects = high
contrast).
1. Assuming a constant (non-adjustable) light source power, identify the part of the microscope
you would adjust to limit the amount of light entering the microscope.
Select all that apply.
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BIOD 171 Essential Microbiology Portage Learning Module

3 Exam Questions and Answers

Question 1

. A nanometer is defined as: A. 10 -^3 B. 10 -^6 C.10-^9 D. 10 -^12 C. A nanometer is defined as one-billionth of a meter.

  1. True or False: A nanometeris longer than a micrometer. False. A nanometer is 1,000 times smaller than a micrometer.
  2. Resolution and contrast are two critical factors that influence your ability to see an object. Explain each. Resolution refers to the distance between two objects at which the objects still can be seen as separate. Poor or low resolution means two (or more) objects may appear as one.
  • Contrast is the difference in light absorbance between two objects. Poor contrast gives a high background and makes the visualization of multiple objects difficult. For instance, tryingto identify 2 dark colored objects at night (low light = low contrast) versus the same 2 objectsin the middle of a sunny afternoon (bright light against 2 dark objects = high contrast).
  1. Assuming a constant (non-adjustable) light source power, identify the part of the microscope youwould adjust to limit the amount of light entering the microscope. Select all that apply.

A. Objective B. Condenser C. Iris diaphragm D. Eye piece C. The iris controls the amount of light that passes through the sample and into the objectivelens. Thus, it can be adjusted (opened or closed) to alter the amount of light.

  1. What is the total magnification (relative to your eye) of a sample imaged with a 60x objective and a 10x eyepiece? Show your math. 60 x 10 = 600x magnification
  2. True or False: Staining is often required to image a cell that is adherent and flat (thin). True. Adherent, flat and unstained cells are almost invisible due to the limits on both resolutionand contrast. Therefore, cell staining is often required to adequately image the sample.
  3. Which of the following could NOT be seen clearly by the unaided eye? Select all that apply. A. Bacteria with diameter of 24 μm B. Protozoa with diameter of 150 μm C. Virus with a diameter of 0.2 μm D. Skin cell with diameter of 1500 μm A and C. The unaided eye can, on average, clearly resolve objects > 100 μm
  4. Label the following unmarked microscope components (numbered arrows) by matching it with thecomponents provided (letters).
  1. This type of microscope utilizes ultraviolet (UV) light to illuminate stained objects.vFluorescence
  2. This type of microscope uses a specialized condenser and objective to amplify the slight differences between cells and background.vPhase-Contrast
  3. This type of microscope enhances contrast between specimen and background but does not permit the visualization of intracellular structures. Dark Field
  4. This type of microscope uses neither halogen nor UV light sources but rather lasers to illuminatestained cells in high resolution. Confocal
  5. Identify what type of electron microscope was used to capture the following image and explainyour choice.

The above image is captured via a Transmission Electron Microscope (TEM). Even at 20nmresolution (inset image), subcellular substructures are still visible. The image lacks the outside ‘shell’ only appearance of SEM.

  1. Gram-Positive cells appear in color due to a peptidoglycan layer in the cell wall. Purple; Thick
  2. Gram-Negative cells appear in color due to a peptidoglycan layer in the cell wall. Pink; Thin
  3. True or False: Following the decolorization step of the Gram stain, Gram-Negative bacteria will appear colorless. True. Even together, the LPS and thin peptidoglycan layer are unable to retain the crystal violet dye during decolorization.
  4. Name one substance capable of chemically fixing cells to a slide. Any of the following are true:Paraformaldehyde, ethanol or methanol.
  5. You want to observe the size and shape of a cell. What is the easiest staining technique that you could perform? Name at least one dye you would use during this process. Simple stain. You coulduse any of the following: methylene blue, crystal violet, safranin or fuschin.