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BIO 120.01 > Microscopy and Staining > Flashcards

Microscopy and Staining Flashcards

1
Q

What is magnification?

A

The capacity of a microscope to enlarge an image uising the lenses in the microscope.

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2
Q

What is resolution?

A

It allows to distinguish between two adjacent objects and separate them into two objects.

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3
Q

What is resolution limit?

A

The limit depends on the numerical aperture. It is highly dependent on light and power.

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4
Q

What is contrast?

A

It allows to differentiate between the cell and surroundings in absorbing or scattering light.

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5
Q

Oragnisms are (blank) so there is a need to create (blank).

A

tranpsarent, contrast

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6
Q

How is contrast achieved?

A

Through various stains and dyes

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7
Q

This microscope uses illumination from visible light.

A

Bright Field Microscope

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8
Q

Light in BFM may come from (blank) or (blank).

A

external, microscope

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9
Q

How do we see images in BFM?

A

Light source concentrates light into the condenser, projecting it to the objective lens

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10
Q

How does the condenser work in Dark Field Microscope?

A

Condenser prevents light from passing through. Light is directed to the sides of the specimen.

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11
Q

What is the Dark Field Microscope used for?

A

To observe motility

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12
Q

Phase Contrast Microscope relies on?

A

The refractive indices of cells (ability of cells to alter the speed of light) and the background

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13
Q

True or false. Cells have unique refractive indices, resulting to different brightness.

A

True

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14
Q

How does Phase Contrast Microscope work?

A

Amplified by the phase ring in the condenser

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15
Q

What is Phase Contrast Microscope used for?

A

Used for living organisms that cannot be stained

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16
Q

The Nomarski Microscope is also called?

A

Differential Interference Contrast [DIC]

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17
Q

This microscope uses a polarizer in the condenser to produce polarized light.

A

Nomarski

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18
Q

In Nomarski, light passes through a (blank) generating (blank) with different refractive indices.

A

prism, 2 beams

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19
Q

How does Nomarski work?

A

Light interfere w/ one another, creating a 3D optical effect, enhancing the subtle differences in cell structures

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20
Q

True or false. Nomarski cannot show internal cell structures and is used to observe motility.

A

False. Nomarski is used for unstained cells, revealing internal cell structures invisible by BFM without staining.

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21
Q

Fluorescence Microscope uses (blank) that can activate fluorescence molecules in cells.

A

UV light

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22
Q

Example of naturally occuring activated fluorescence

A

chlorophylls

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23
Q

Examples of stain that can activate fluorescence?

A

fluorochromes, DAPI

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24
Q

How does Fluorescence Microscope work?

A

Filters eliminate other wavelengths of light other than the fluorescent light

25
Q

What is the use of Fluorescence Microscope?

A

Visualizing certain cell structures

26
Q

Computer assembles multiple layers to create a single high-resolution, 3D image.

A

Confocal Scanning Laser

27
Q

Best for thick specimens.

A

Confocal Scanning Laser

28
Q

In Confocal Scanning Laser, dead cells are stained (blank) and viable are stained (blank).

A

red, green

29
Q

Electron microscopy uses (blank) as lenses.

A

electromagnets

30
Q

Electron microscope is opened in a (blank).

A

vaccum

31
Q
  • allows you to visualize the internal cell structures
  • magnification up to 500,000x

proteins & nucleic acids are visible

A

Transmission Electron Microscope

32
Q

Samples in TEM are stained using (blank) with (blank) to improve contrast.

shadow-casted

A

atom, high atomic weights

33
Q
  • used to visualize external structures or surfaces of the cell
  • magnification up to 100,000x
A

Scanning Electron Microscope

34
Q

Specimen in SEM is usually coated in (blank).

A

thin sheet of heavy metal

35
Q

How does SEM work?

A

Electron beams scan the surface then direct it back to the computer

36
Q

Tiny stylus positioned extremely close to the specimen surface

A

Atomic Force Microscopy

37
Q

Which microscope can be used at the surface of live specimens?

A

Atomic Force Microscopy

38
Q

Wet mounts don’t need to be stained with dyes. What is used to see the specimen?

A

hay infusion, pond water

39
Q

Addition of (blank) can create a viscous environment to slow down movement.

A

carboxymethylcellulose

40
Q

If carboxymethylcellulose is not avaiable, what can we use to create “tents” to trap organisms incone area?

A

cotton fiber

41
Q

True or false. E. coli is motile.

A

True

42
Q

What will the cell arrangement look like if you smeared too much?

A

Cell arrangement won’t be too visible

cells will be on top of each other

43
Q

What will the cell arrangement look like if your smear is too thin?

A

Arrangement will be scattered everywhere

44
Q

Examples of positively charged dyes:

A

crystal violet, methylene blue, safranin

45
Q

Why do we need to heat fix?

A

To attach bacteria on slide

46
Q

True or false. Positive staining include acidic dyes.

A

False. Positive staining dyes are basic/cationic.

methylene blue, crystal violet, safranin

47
Q

Example of acidic dyes:

A

nigrosine black, India ink, and Congo red

48
Q

True or false. We need to heat fix everytime.

A

False. No need for heat fixing if dye is negative.

Organisms are heat-sensitive

49
Q

Three different types of staining:

A

simple, structural, differential

50
Q

Highlights certain structures of the cell

A

structural staining

51
Q

Schaeffer-Fulton Method is also called?

A

Endospore staining

52
Q

Primary stain for endospore staining

A

Malachite green

53
Q

Endospore wall becomes permeable to malachite green due to (blank).

A

steaming

54
Q

Gram staining differentiate betwen (blank) and (blank).

A

positive, negative

55
Q

Acid fast staining is also called?

A

Ziehl-neelsen Stain Method

56
Q

You can see the presence of (blank) in Ziehl-neelsen Stain Method

A

mycolic acids

retains primary stain: carbolfuchsin

57
Q

Primary stain for Ziehl-Neelsen

A

carbolfuchsin

58
Q

Counter stain of Ziehl-Neelsen

A

methylene blue

BIO 120.01 (12 decks)

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