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Fill in the Blank Questions
1. The __________ is the point at which a lens focuses parallel beams of light.
focal point
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.01.02 Correlate lens strength and focal length
Section: 02.01
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
2. The __________ is the distance between the center of a lens and the point at which it
focuses parallel beams of light.
focal length
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.01.01 Relate the refractive indices of glass and air to the path light
takes when it passes through a prism or convex lens
Section: 02.01
Chapter 02 – Microscopy
2-2
True / False Questions
3. Light rays are refracted (bent) when they cross the interface between materials with
different refractive indices.
TRUE
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.01.02 Correlate lens strength and focal length
Section: 02.01
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Multiple Choice Questions
4. Light rays are refracted (bent) when they cross the interface between materials with
different refractive indices.
A. differential interference contrast
B. dark field
C. phase-contrast
D. confocal
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Blooms Level: 2. Understand
Learning Outcome: 02.01.01 Relate the refractive indices of glass and air to the path light
takes when it passes through a prism or convex lens
Section: 02.01
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-3
5. Confocal microscopes exhibit improved contrast and resolution by
A. illumination of a large area of the specimen.
B. blocking out stray light with an aperture located above the objective lens.
C. use of light at longer wavelengths.
D. use of ultraviolet light to illuminate the specimen.
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Learning Outcome: 02.02.02 Predict the relative degree of resolution based on light
wavelength and numerical aperture of the lens used to examine a specimen
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
6. A 30 objective and a 20 ocular produce a total magnification of
A. 230.
B. 320.
C. 50.
D. 600.
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 3. Apply
Learning Outcome: 02.02.01 Evaluate the parts of a light microscope in terms of their
contributions to image production and use of the microscope
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-4
7. A 45 objective and a 10 ocular produce a total magnification of
A. 900.
B. 55.
C. 450.
D. 145.
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 3. Apply
Learning Outcome: 02.02.01 Evaluate the parts of a light microscope in terms of their
contributions to image production and use of the microscope
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
8. A microscope that exposes specimens to ultraviolet, violet, or blue light and forms an
image with the light emitted at a different wavelength is called a __________ microscope.
A. phase-contrast
B. dark-field
C. scanning electron
D. fluorescence
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.02.03 Create a table that compares and contrasts the various types of
light microscopes in terms of their uses, how images are created, and the quality of images
produced
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-5
9. Immersion oil can be used to increase the resolution achieved with some microscope lenses
because it increases the __________ between the specimen and the objective lens.
A. optical density
B. refractive index
C. optical density and refractive index
D. neither optical density nor refractive index
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Blooms Level: 2. Understand
Learning Outcome: 02.01.01 Relate the refractive indices of glass and air to the path light
takes when it passes through a prism or convex lens
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
True / False Questions
10. A substage condenser is used to focus light onto the specimen, which increases the
resolution of a light microscope.
TRUE
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.05.02 Evaluate light microscopy, electron microscopy, and scanning
probe microscopy in terms of their uses, resolution, and the quality of the images create
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-6
Fill in the Blank Questions
11. The __________ is the distance between the specimen and the objective lens when the
specimen is in focus.
working distance
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Blooms Level: 2. Understand
Learning Outcome: 02.02.01 Evaluate the parts of a light microscope in terms of their
contributions to image production and use of the microscope
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
12. The useful magnification of a light microscope is limited by the ___________ of the light
source being utilized.
wavelength
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Blooms Level: 2. Understand
Learning Outcome: 02.02.02 Predict the relative degree of resolution based on light
wavelength and numerical aperture of the lens used to examine a specimen
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-7
13. The special dyes used in fluorescence microscopy that absorb light at one wavelength and
emit light at a different wavelength are called __________.
fluorochromes
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.02.02 Predict the relative degree of resolution based on light
wavelength and numerical aperture of the lens used to examine a specimen
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
14. In order to view a specimen with a total magnification of 400, a __________ objective
must be used if the ocular is 10.
40
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 3. Apply
Learning Outcome: 02.02.01 Evaluate the parts of a light microscope in terms of their
contributions to image production and use of the microscope
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-8
True / False Questions
15. Confocal microscopes, in combination with specialized computer software, can be used to
create three-dimensional images of cell structures.
TRUE
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Blooms Level: 1. Remember
Learning Outcome: 02.02.03 Create a table that compares and contrasts the various types of
light microscopes in terms of their uses, how images are created, and the quality of images
produced
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
16. A light microscope with an objective lens numerical aperture of 0.65 is capable of
allowing two objects 400 nm apart to be distinguished when using light with a wavelength of
420 nm.
TRUE
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Blooms Level: 3. Apply
Learning Outcome: 02.02.02 Predict the relative degree of resolution based on light
wavelength and numerical aperture of the lens used to examine a specimen
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
17. Resolution decreases when the wavelength of the illuminating light decreases.
FALSE
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Learning Outcome: 02.02.02 Predict the relative degree of resolution based on light
wavelength and numerical aperture of the lens used to examine a specimen
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-9
18. Immersion oil is used to prevent a specimen from drying out.
FALSE
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
19. It is possible to build a light microscope capable of 10,000 magnification, but the image
would not be sharp because resolution is independent of magnification.
TRUE
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Blooms Level: 2. Understand
Learning Outcome: 02.01.02 Correlate lens strength and focal length
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
20. Immersion oil increases the amount of light passing through a specimen and entering the
objective lens.
TRUE
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.01.01 Relate the refractive indices of glass and air to the path light
takes when it passes through a prism or convex lens
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-10
Multiple Choice Questions
21. If the objective lenses of a microscope can be changed without losing focus on the
specimen, they are said to be
A. equifocal.
B. totifocal.
C. parfocal.
D. optifocal.
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.02.01 Evaluate the parts of a light microscope in terms of their
contributions to image production and use of the microscope
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
22. An instrument that magnifies slight differences in the refractive index of cell structures is
called a (n) __________ microscope.
A. phase-contrast
B. electron
C. fluorescence
D. densitometric
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Learning Outcome: 02.02.03 Create a table that compares and contrasts the various types of
light microscopes in terms of their uses, how images are created, and the quality of images
produced
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-11
23. The instrument that produces a bright image of the specimen against a dark background is
called a (n) __________ microscope.
A. phase-contrast
B. electron
C. bright-field
D. dark-field
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
Blooms Level: 2. Understand
Learning Outcome: 02.02.03 Create a table that compares and contrasts the various types of
light microscopes in terms of their uses, how images are created, and the quality of images
produced
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
24. As the magnification of a series of objective lenses increases, the working distance
A. increases.
B. decreases.
C. stays the same.
D. cannot be predicted.
ASM Objective: 08.05 Use appropriate microbiological and molecular lab equipment.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.02.01 Evaluate the parts of a light microscope in terms of their
contributions to image production and use of the microscope
Section: 02.02
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-12
25. Prior to staining, smears of microorganisms are heat-fixed in order to
A. allow eventual visualization of internal structures.
B. ensure removal of dust particles from the slide surface.
C. attach it firmly to the slide.
D. create small pores in cells that facilitates binding of stain to cell structures.
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Objective: 08.02 Use aseptic and pure culture techniques to enrich for and isolate
microorganisms.
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.03.01 Recommend a fixation process to use when the microbe is a
bacterium or archaeon and when the microbe is a protist
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
26. Acid-fast organisms such as Mycobacterium tuberculosis contain __________ constructed
from mycolic acids in their cell walls.
A. proteins
B. carbohydrates
C. lipids
D. peptidoglycan
ASM Objective: 02.02 Bacteria and Archaea have specialized structures (e.g. flagella,
endospores, and pili) that often confer critical capabilities.
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-13
27. In the Gram-staining procedure, the primary stain is
A. iodine.
B. safranin.
C. crystal violet.
D. alcohol.
ASM Topic: Module 02 Structure and Function
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 1. Remember
Learning Outcome: 02.03.03 Compare what happens to Gram-positive and Gram-negative
bacterial cells at each step of the Gram-staining procedure
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
28. In the Gram-staining procedure, the decolorizer is
A. iodine.
B. safranin.
C. crystal violet.
D. ethanol or acetone.
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-14
29. In the Gram-staining procedure, the counterstain is
A. iodine.
B. safranin.
C. crystal violet.
D. alcohol.
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
30. In the Gram-staining procedure, the mordant is
A. iodine.
B. safranin.
C. crystal violet.
D. alcohol.
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-15
31. After the primary stain has been added but before the decolorizer has been used, grampositive organisms are stained __________ and gram-negative organisms are stained
__________.
A. purple; purple
B. purple; colorless
C. purple; pink
D. pink; pink
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.03 Compare what happens to Gram-positive and Gram-negative
bacterial cells at each step of the Gram-staining procedure
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
32. After the decolorizer has been added, gram-positive organisms are stained __________
and gram-negative organisms are stained __________.
A. purple; purple
B. purple; colorless
C. purple; pink
D. pink; pink
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
ASM Topic: Module 08 Microbiology Skills
Blooms Level: 2. Understand
Learning Outcome: 02.03.03 Compare what happens to Gram-positive and Gram-negative
bacterial cells at each step of the Gram-staining procedure
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-16
33. After the secondary stain has been added, gram-positive organisms are stained
__________ and gram-negative organisms are stained __________.
A. purple; purple
B. purple; colorless
C. purple; pink
D. pink; pink
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.03 Compare what happens to Gram-positive and Gram-negative
bacterial cells at each step of the Gram-staining procedure
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
34. If the decolorizer is left on too long in the Gram-staining procedure, gram-positive
organisms will be stained __________ and gram-negative organisms will be stained
__________.
A. purple; blue
B. purple; colorless
C. purple; pink
D. pink; pink
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 3. Apply
Learning Outcome: 02.03.03 Compare what happens to Gram-positive and Gram-negative
bacterial cells at each step of the Gram-staining procedure
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-17
35. If the decolorizer is not left on long enough in the Gram-staining procedure, gram-positive
organisms will be stained __________ and gram-negative organisms will be stained
__________.
A. purple; purple
B. purple; colorless
C. purple; pink
D. pink; pink
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 3. Apply
Learning Outcome: 02.03.03 Compare what happens to Gram-positive and Gram-negative
bacterial cells at each step of the Gram-staining procedure
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
36. Which of the following is considered to be a differential staining procedure?
A. Gram stain.
B. Acid-fast stain.
C. both Gram stain and Acid-fast stain.
D. Leifson’s flagella stain.
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-18
37. Basic dyes such as methylene blue bind to cellular molecules that are
A. hydrophobic.
B. negatively charged.
C. positively charged.
D. aromatic.
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
38. The Schaeffer-Fulton procedure is used to stain
A. flagella.
B. fat deposits.
C. endospores.
D. DNA of chromosomes.
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-19
True / False Questions
39. Gram staining divides bacterial species into roughly two equal groups.
TRUE
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.03 Compare what happens to Gram-positive and Gram-negative
bacterial cells at each step of the Gram-staining procedure
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
40. Negative staining facilitates the visualization of bacterial capsules which are intensely
stained by the procedure.
FALSE
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
Chapter 02 – Microscopy
2-20
41. Negative staining with India ink can be used to reveal the presence of capsules that
surround bacterial cells.
TRUE
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.02 Plan a series of appropriate staining procedures to describe an
unknown bacterium as fully as possible
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms
42. Mordants increase the binding between a stain and specimen.
TRUE
ASM Objective: 08.01 Properly prepare and view specimens for examination using
microscopy (bright field and, if possible, phase contrast).
ASM Topic: Module 02 Structure and Function
Blooms Level: 2. Understand
Learning Outcome: 02.03.03 Compare what happens to Gram-positive and Gram-negative
bacterial cells at each step of the Gram-staining procedure
Section: 02.03
Topic: Tools and Methods of Culturing, Classifying, and Identify Microorganisms