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Human Movement Dissected > Microscopy > Flashcards

Microscopy Flashcards

1
Q

What are the types of unstained light microscopy techniques?

A

Bright Field, Phase Contrast and Differential Interference contrast

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2
Q

What is Bright Field?

A

Normal, no special techniques

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3
Q

What is Phase Contrast?

A

Uses same size phase condenser to enhance the differences between wavelengths. This is a very simple technique

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4
Q

What is Differential Interference Contrast?

A

Very complicated

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5
Q

What is Kohler Illumination?

A

The technique for optimising resolution and creating even sample illumination

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6
Q

What is sample fixation?

A

Preserving the structure and contents of tissue.

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7
Q

What is an example of a fixation agent?

A

Formaline

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8
Q

What is the dehydration step?

A

Dehydration is done to remove all liquid and ‘dry’ the sample. This often uses ethanol.

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9
Q

What is used to remove the dehydrating agents?

A

A clearing agent such as chloroform

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10
Q

What is embedding?

A

Solidifying a sample so that it can be cut

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11
Q

What are the two main types of embedding?

A

OCT freezing and molten parrafin wax.

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12
Q

What are the benefits of using OCT freezing?

A

Allows the antigenic properties to be preserved- used in immunohistochemistry.

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13
Q

What are the benefits of wax embedding?

A

Allows the structure of tissue to be retained- used in pathology

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14
Q

What is a cryostat?

A

Freezing and microtome machine (5-10um)

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15
Q

How thin does a microtome cut to?

A

4-30um

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16
Q

What does haematoxylin stain?

A

Nuclei dark blue and all other structures pink/purple

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17
Q

What does Safranin O and Fast Green stain?

A

Cartilage red and muscle green/blue

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18
Q

When would you use Safranin O/Fast Green stain?

A

In the study of osteoarthritis

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19
Q

What does Masson trichrome stain?

A

Collagen blue, nuclei black and muscle/cytoplasm/keratin red

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20
Q

What does Vanhoff Gierson stain?

A

Elastic fibres and nuclei blue/black, collagen red and other structures yellow

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21
Q

When would you use Masson trichrome stain?

A

Liver, Kidney or tumor biopsy

22
Q

When would you use Vanhoff Gierson stain?

A

Elastic structures such as blood vessels and in tumour invasion determination

23
Q

What is a flurophore?

A

A substance that emits light at a different wavelength to that at which it absorbed it

24
Q

What is stoke shift?

A

Difference between the absorbed and emitted wave lengths

25
Q

What is a filter cube?

A

A cube which allow specific wave length to hit the target and a detector to absorb the emission from the target without interference from the original source.

26
Q

What is direct staining?

A

An antibody bound to the stain directly binds to the target.

27
Q

What is indirect staining?

A

The antibody bound to the stain binds to another antibody, which itself has bound to the target

28
Q

What does Wheat Germ Agglutinin stain?

A

Plasma membrane

29
Q

What does Phalloidin stain?

A

Actin

30
Q

What does DAPI stain?

A

Double-stranded DNA

31
Q

What is the extinction coefficient?

A

The intrinsic property of a substance to absorb/reflect certain waves lengths

32
Q

Does the Formaldehyde preserve structure or antigenicity?

A

Structure

33
Q

Does the Methanol fixing agent preserve structure or antigenicity?

A

Antigenicity

34
Q

What is permabilisation?

A

‘poking’ holes in the cell to allow the stain to enter. An example of this is Tritin X.

35
Q

What is blocking?

A

Blocking undesirable antibody binding site

36
Q

What is often used as a blocking agent

A

Bovine Serum Albumin

37
Q

What is confocal microscopy?

A

Using a thin ‘slice’ of the visual field reduces light interference and increases resolution. It also allows for 3D images to be constructed

38
Q

What is widefield?

A

Normal microscopy

39
Q

When do you use transmission electron microscopy?

A

For small samples and to their ultrastructure

40
Q

When do you use a scanning electron microscope?

A

For larger samples and to see their surface in detail

41
Q

What is a conventional SEM electron source?

A

A plate is heated up until it produces electrons. This is more common and simple, but as the electrons are not as focused it cannot give the ‘zoom’ of the other type (50,000)

42
Q

What is a field emission SEM electron source?

A

A strong electromagnetic field is created with a current to push electrons into the beam. The electron beam is much more powerful and therefore can increase the zoom and resolution (1,000,000x)

43
Q

What is backscatter vs non-backscatter?

A

Non-backscatter uses the secondary electrons emitted from the sample to create an image whereas backscatter uses the electrons which are reflected from the sample to create the image

44
Q

What must happen when samples are viewed under SEM High Vacuum?

A

They must be gold-palladium plated

45
Q

What does Variable pressure SEM allow for?

A

Uncoated fresh samples to be viewed

46
Q

What is the gold-palladium coating known as?

A

The sputter coat

47
Q

Is the sputter coat applied first or second?

A

It is the secondary fixing agent

48
Q

When fixating TEM, which methods can be used?

A

Chemical fixation or critical point freezing fixation

49
Q

What is an ultramicrotome?

A

A microtome that sections to 90nm

50
Q

With TEM, what can be used as the primary fixator?

A

2.5% glutaraldehyde

51
Q

In TEM, what can be used as the secondary fixator?

A

1% Osmium tetroxide

52
Q

What is used for critical point drying?

A

Hexamethyldisilizane

Human Movement Dissected (3 decks)

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