Microscopy Flashcards

1
Q

what is a light field microscope?

A

most common, observes stained sections, cells must be fixed

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2
Q

what is a phase contrast microscopy?

A

observes living cells, black and white image, amplifies the difference in RI

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3
Q

what cells is phase contrast microscopy good for?

A

live and unstained cells

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4
Q

what is differential interference contrast microscopy?

A

similar to phase contrast but greater resolution of 3D appearance

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5
Q

what is dark field microscopy?

A

illuminates cells and tissues against a dark background to allow contrast

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6
Q

what is fluorescent microscopy?

A

uses fluorescent molecules to observe structures

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7
Q

what is confocal microscopy?

A

uses lasers and optical modifications to focus on a single plane

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8
Q

what pieces of tissue are good for staining?

A

thin sections that are transparent to light

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9
Q

what is a vital dye?

A

can penetrate cells and assess culture viability

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10
Q

what is dye exclusion?

A

dye that cannot penetrate viable cells, such as trypan blue

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11
Q

what is dye uptake?

A

dye taken up by a living cell such as methylene blue

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12
Q

what does haematoxylin do?

A

stains nuclei black and cytoplasm pink

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13
Q

how does enzyme histochemistry work?

A

tissue incubated with substrates and when the enzymes react, a coloured product is produced

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14
Q

what is autoradiography?

A

makes use of radioactive emissions blackening a photographic plate

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15
Q

what is an example of autoradiography?

A

precursor substances such as H or C injected in vivo, when in contact with photographic plate, the radioactive isotope causes the silver grains to blacken

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16
Q

what is interference microscopy?

A

one beam through the specimen and one not, light through the specimen has a phase change, causing a difference in refractive index giving fringe pattern

17
Q

what is a use of interference?

A

can be used for quantitative measures such as dry weight

18
Q

what is a polarising microscope?

A

specimen placed between crossed polaroids and some areas where polarisation is rotated by the birefringence of the specimen, it is brighter than the background

19
Q

what is birefringence?

A

property of a material where the RF of a material depends on the polarisation of light

20
Q

what is UV microscopy?

A

certain substances strongly absorb UV light so detected if illuminated by UV light

21
Q

what is direct immunofluorescence?

A

specific antibodies to the antigen are injected which are labelled with fluorescent dye

22
Q

what is indirect immunofluorescence?

A

primary antibody injected and allowed to bind and secondary labelled antibody that is complementary is injected

23
Q

what may cause artefacts to be formed?

A

swelling or shrinking due to tissue fixation, inadequate staining, enzyme action, incomplete dehydration

24
Q

what is the resolution equation?

A

D = 0.61 x wavelength/rf of air x sin x alpha

25
Q

what is the alpha in the resolution equation?

A

angular aperture