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Cell Biology > Module 06: Microscopy > Flashcards

Module 06: Microscopy Flashcards

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0
Q

What do you during when you fix cells?

A

Covalently crosslink macromolecules to each other

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1
Q

What is the purpose of dehydration when you fix cells?

A

Prevention of microbial growth and hydrolysis

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2
Q

What is the purpose of permeabilizing cells during fixation?

A

Degrades the membrane, allows access of antibodies and stains

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3
Q

What does a microtome do?

A

It cuts tissue into sections thin enough that light can transmit rough

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4
Q

What are the steps of preparing a thick specimen?

A

Fixing the tissue, dehydration, embedding in block of wax, microtoming

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5
Q

What classes of chemicals are used in chemical fixation?

A

Acids (acetic acid) and aldehydes (formaldehyde)

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6
Q

What processing is required to use phase contrast microscopy?

A

None; cells are unprocessed

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7
Q

How does phase contrast microscopy work?

A

Uses the different in phases between direct (unaltered) and diffracted (altered by specimen) light

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8
Q

What does DIC microscopy stand for?

A

Differential interference microscopy

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9
Q

What is another name for DIC microscopy?

A

Normanski microscopy

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10
Q

Describe the basics of fluorescence.

A

A photon causes an electron to move to an excited state, which then drops to a lower, more stable state. This energy is released as an emitted photon (of a longer wavelength)

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11
Q

What is it called when a photon causes an electron to move to a higher energy state?

A

Excitation

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12
Q

What is it called when an electron drops to a more stable energy state and releases a photon?

A

Emission

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13
Q

excitation (n.)

A

the act of a photon exciting an electron into a higher energy state

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14
Q

How does immunofluoresence work?

A

Fluorescently labeled antibodies bind directly to antigen

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15
Q

How does indirect immunofluoresence work?

A

An unlabeled primary antibody binds to the antigen; secondary antibodies labeled with a fluorescent dye binds to the primary antibodies

16
Q

What does DAPI stain?

A

DNA

17
Q

What does GFP stand for and where does it come from?

A

Green fluorescent protein, and it comes from jellyfish

18
Q

What is special about GFP?

A

It allows us to image and record live cells

19
Q

How is GFP added onto tubulin?

A

DNA coding for GFP is added on to the sequence that encodes for tubulin; the end result is a fusion protein of tubulin and GFP

20
Q

What does TEM stand for?

A

Transmission electron microscopy

21
Q

What are two heavy metals used in TEM?

A

OsO4 and gold

22
Q

What does SEM stand for?

A

Scanning Electron Microscopy

23
Q

What is the preparation process for SEM?

A

Fixation, dehydration, soaked in OsO4, then sprayed with a fine coat of gold

24
Q

What is OsO4?

A

Osmium tetroxide

25
Q

Does OsO4 bind to polar or non-polar molecules?

A

Polar molecules

26
Q

How does negative staining work?

A

Puddle an electron dense solution around a 3D structure

27
Q

What kind of microscopy do you use to look at the result of the “shadowing” process?

A

TEM

28
Q

What is freeze fracture used for?

A

Used for looking inside membranes

29
Q

What type of microscopy is used after freeze fracturing?

A

TEM

30
Q

How big is the average eukaryotic cell?

A

20 µm

31
Q

How big is the average protein?

A

2 nm

32
Q

What type of energy is light?

A

Electromagnetic energy

33
Q

What property of light changes to make the different colors of visible light?

A

The wavelength

34
Q

How does an electron get to a higher orbital?

A

Absorb energy

35
Q

How does an electrons get to a lower orbital?

A

Release energy

36
Q

What is a photon?

A

A “packet” of light

37
Q

How are frequency and wavelength related?

A

The higher the frequency, the shorter the wavelength

38
Q

How are electromagnetic energy and wavelength related?

A

The shorter the wavelength, the higher the energy

Cell Biology (6 decks)

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