CAMAING, CHARMAINE R.

BSED 3-SCIENCE

Answer the following:

1. Which type of microscope would be best to use to observe each of the following?
a. a stained bacterial smear – Compound Light Microscope
b. unstained bacterial cells: the cells are small, and no detail is needed –
Darkfield Microscope
c. unstained live tissue when it is desirable to see some intracellular detail –
Phase-Contrast Microscope
d. a sample that emits light when illuminated with ultraviolet light -
Fluorescence Microscope
e. intracellular detail of a cell that is 1 μm long - Electron Microscope
f. unstained live cells in which intracellular structures are shown in color -
Differential Interference Contrast Microscope

2. Calculate the total magnification of the nucleus of a cell being observed through a
compound light microscope with a 10* ocular lens and an oil immersion lens.
 Ocular lens- 10X; Oil Immersion Lens- 100X=Total Magnification- 1000X

3. The maximum magnification of a compound microscope is (a) 2,000X; that of an

electron microscope, (b) 100,000X. The maximum resolution of a compound
microscope is (c) 0.2μm; that of an electron microscope, (d) 0.0025μm. One
advantage of a scanning electron microscope over a transmission electron
microscope is (e) seeing three-dimensional detail.

4. Why is immersion oil necessary at 1000x but not with the lower power objectives?
 Basically when using lower magnification microscope objective lenses (4x,
10x, 40x) the light refraction is not usually noticeable. However, once you use
the 100x objective lens, the light refraction when using a dry lens is noticeable.
If you can reduce the amount of light refraction, more light passing through the
microscope slide will be directed through the very narrow diameter of a higher
 power objective lens. In microscopy, more light = clear and crisp images. By
placing a substance such as immersion oil with a refractive index equal to that
of the glass slide in the space filled with air, more light is directed through the
objective and a clearer image is observed.

Discussion Board

1. Through what lenses does light pass in a compound microscope?

 Microscopes are equipped with specific lenses that can make microorganisms
visible to the human eye. This instrument has a lens through which a human can
look and observe the object (microorganism). This lens is called as objective and
ocular lens. This remains close to the human eye. So, it is also called an eyepiece. It
is responsible for magnifying the image of the organism under observation. It
usually has 10x the power of magnification. It is a convex lens.

2. What does it mean when a microscope has a resolution of 0.2 nm?

 It means that it can distinguish objects greater than or equal to 0.2nm.

3. What are the advantages of brightfield, darkfield, and phase-contrast microscopy?

 Brightfield, darkfield, and phase contrast are the most common label-free contrast
modes used in optical microscopy. Brightfield imaging is most suitable for observing
samples with strong absorption. Darkfield imaging provides good contrast for
subresolution features, since it only captures high-angle scattered light. Phase
contrast is used for unstained and transparent biological samples, allowing
visualization of shape and density variations.

4. Why is a DIC micrograph brightly colored?

 Differential interference contrast (DIC) microscopes (also known as Nomarski
optics) are similar to phase-contrast microscopes in that they use interference
 patterns to enhance contrast between different features of a specimen. In a DIC
microscope, two beams of light are created in which the direction of wave
movement (polarization) differs. Once the beams pass through either the specimen
or specimen-free space, they are recombined and effects of the specimens cause
differences in the interference patterns generated by the combining of the beams.
This results in high-contrast images of living organisms with a three-dimensional
appearance. These microscopes are especially useful in distinguishing structures
within live, unstained specimens.

5. Why won’t other bacteria fluoresce in the FTA-ABS test?

 Because it is not the same bacteria.

6. What are the advantages of confocal microscopy?

 It shows the physiology of the cell.

7. What are the differences between TPM and confocal microscopy?

 Two-Photon Microscope (TPM) is a light microscope that uses fluorescent stains
and long wavelengths of light, while confocal microscopy is a light microscope that
uses fluorescent stains and laser to make two- and three-dimensional images.

8. How are brightfield, darkfield, phase-contrast, and fluorescence microscopy similar?

 These microscopes fall under light microscopy which uses lenses to focus light on a
specimen to produce an image.

9. What is the advantage of super-resolution light microscopy?

 Super-resolution microscopy achieves better resolution at greater depths by
controlling the modulation of the excitation light as opposed to light excitation in a
uniform pattern.
 10. What is the principal use of SAM?
 Scan Acoustic Microscopy (SAM) use sound wave to construct image to examining
cancel cell attached to one surface.

11. How do TEM and SEM images of the same organism differ?
 TEM shows the layers of the cell, while, SEM shows the surface.

12. Why do electron microscopes have greater resolution than light microscopes?
 Electron microscopes differ from light microscopes in that they produce an image of
a specimen by using a beam of electrons rather than a beam of light. Electrons have
much a shorter wavelength than visible light, and this allows electron microscopes
to produce higher-resolution images than standard light microscopes.

13. What is the principle employed in scanned-probe microscopy?

 Scanned-probe microscopy (SPM) is use to study molecules inside the cell such as –
DNA.

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