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BB10005- Cell Biology > Light microscopy (lecture 2) > Flashcards

Light microscopy (lecture 2) Flashcards

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1
Q

Structures in which we can see with a light microscope?

A 
frog egg 
eu cells 
nucleus 
most bacteria 
mito
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2
Q

Resolution is determined by?

A

wavelength of radiation source used for illumination

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3
Q

Size in which is able to be seen with the naked eye (good eye sight)

A

0.2mm

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4
Q

Magnify an image what is needed?

A

Magnifier or lens

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5
Q

Cornea?

A

focus light onto the retina

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6
Q

Magnification?

A

Magnification: the ratio of the size of the image to that of the object.

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7
Q

Resolution?

A

Resolution: the clarity of the image; illumination and quality of the optics.

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8
Q

Contrast?

A

Contrast: the contrast between the lightest and darkest areas of the sample.

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9
Q

How to calculate total magnification?

A

Magnification of the objective lens and multiple by the eyepiece

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10
Q

Robert hooke? Time, what he discovered and how

A

17th century
- the cell
cork sections - tree

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11
Q

Antony van leeuwenhoek?

A

Collaborated with Hooke in which he improved the production of the lenses and observed different cell types (eg red blood cells)

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12
Q

Specimen holder?

A

Now known as a stage

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13
Q

In the 17th century, a lens was?

A

water flask

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14
Q

Zeiss?

A

1866
lens better
less diffraction
greater level of detail (improved the resolution)

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15
Q

Prism allows

A

to sit at a desk whilst observing structures under a microscope

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16
Q

If we adjust the condenser lens?

A

locus light onto a specimen

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17
Q

Objective lens

A

collects cone of light rays to create an image

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18
Q

5 different types of light microscopy?

A

1) Bright Field
2) Phase contrast
3) Differentail inference contrast (DIC)
4) Fluorescence
5) Confocal

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19
Q

Bright field miscopy image type?

A

Poor images
not much contrast
a lot of light passes through cells

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20
Q

Phase contrast, (high contrast due to?)

A

The diffracted and un-diffracted rays give rise to a change in brightness.

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21
Q

Can we see mitotic divisions occurring in phase contrast microscopy?

A

yes

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22
Q

Light first moves though _____ before passes onto the specimen phase contrast microscopy?

A

Condenser

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23
Q

From the light source, the light then passes through the _________ and creates?

A

Annulgar ring&raquo_space; creates a hollow cone of light

24
Q

From the angular ring, light passes through ______(phase contrast microscopy?)

A

the specimen

25
Q

Diffraction depends on (phase contrast microscopy?)

A

Density of the specimen

26
Q

Where is light collected (phase contrast microscopy?)

A

Phase ring

27
Q

How is a greater detail achieved (phase contrast microscopy?)

A

Change in amplitude

28
Q

DIC stands for?

A

Differentail inference contrast

29
Q

DIC uses?

A

Polarised light

30
Q

What kind of prism is required and purpose? (DIC)

A

Beam spliter prism

Generating two beams of light which travel close to eachother

31
Q

If one of the beams is diffracted by the specimen (DIC)??

A

when they are re-combined by the top Wollaston prism they will interfere and generate contrast ranging from black to white

32
Q

If the beams are not diffracted by the specimen (DIC)

A

They are not recombined and produces a pale grey colour

33
Q

Wollaston prism is use for which light miscopy method and what is it?

A

DIC

- 2 quartz wedges, causes a beam of polarised light into 2 beams which travel close to eachother

34
Q

Single cells can be ______ ______

A

viewed directly

35
Q

Tissues may need ________

A

Sectioning

36
Q

Detection of cellular components or cells types may need?

A

Dyes or chromogenic enzyme substrates

37
Q

Example of plant roots cells becoming stained

A

plants produce phenolic compounds if they become damaged, which can be died and detected

38
Q

Lignin in cells walls ???

A

can be dyed and detected

39
Q

example of enzymes being used as dyes

A

Artificial substrate reveals protease activity from apathogenic fungus infecting insect cuticle.

Apply a substrate»> colour change when in the presence of proteases

can see where the fungus is localised

40
Q

Dyes also allow the detection of

A

life or death in cells

41
Q

Example of neutral red dye use??

A

in dead cells (infected by a fungi) dye is not retained by vacuole (as the vacuole membrane is damaged)

Living cells take up dye

42
Q

Fluorescence is the property of??

A

Fluorescence is the property of absorbing light of a particular wavelength and then emitting light of a different wavelength.

43
Q

Visible light is defined by?

A

Visible light is usually defined as having wavelengths in the range of 400–700 nm.

44
Q

What type of filters used for Fluorescence light miscopy ??

A

Selective filters (illuminate at a particular wavelength of light)

45
Q

First light passes through ?? Causes??

A

Filter (particular wavelength of light produced)

Excitement of a fluorescent compound

46
Q

Light is then focused (from filter) Fluorescence light miscopy

A

Onto the objective lens

47
Q

From the objective lens.. light _____ (Fluorescence light miscopy )

A

light passes through a beam splitter

48
Q

How is data viewed (Fluorescence light miscopy)

A

By eye or computer

49
Q

Applications of Fluorescence light miscopy

A

Visualising organelles within cells e.g. by using immunofluorescence microscopy.

locations of molecules within cells (multiple probes multiple structures)

50
Q

Confocal microscopy purpose?

A

Increases resolution

  • standard fluorescence microscopy
51
Q

Confocal microscopy research options ?

A

1) co-localisation

2) intra-cellular studies
3) examination of thick specimens
4) studies where 3-D structure of the sample is important.

52
Q

Confocal microscopy allows for?

A

Optical sections to be made without actually having to form sections within a specimen

53
Q

Image volume for confocal microscopy?

A

DEEP

54
Q

where is light collected from confocal miscopy?

A

Exclusively collect light from the focal plane

55
Q

GFP derived from?

A

Aequorea Victoria- GFP

56
Q

Expression profiles allow??

A

see where a gene is localised

BB10005- Cell Biology (6 decks)

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