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Mbb231 > Microscopy > Flashcards

Microscopy Flashcards

1
Q

Thinnest picture is by

A

Scanning Electron Microscopy

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2
Q

Light microscope kind

A

1- phase contrast
2- fluorescence microscope

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3
Q

Electron microscopes kind

A

1- transmission electrons
2- scanning electrons

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4
Q

Three elements needed for imaging

A

1- source of illumination
2- speciemen to be examined
3- system of lenses

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5
Q

Condenser lens

A

In front of light source

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6
Q

Objective lens

A

Closest lense to object

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7
Q

Ocular lens

A

Magnify image created by objective lense

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8
Q

Magnification

A

Size of the image vs object

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9
Q

Empty magnification

A

Maginified without resolution

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10
Q

Resolving power is

A

Resolution

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11
Q

Resolution allows object to

A

Be distinguished

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12
Q

Resolution

A

Minimum distance two points can be apart and remain apart

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13
Q

Magnification consider

A

1- refraction index of lens
1- medium that sample is immersed in
2- focal length of lens

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14
Q

Resolution considers

A

1- wavelength of illumination
2- refraction index
3- angular aperture

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15
Q

Longer wavelength

A

Larger object observed

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16
Q

Shorter wavelength

A

Smaller object obseved

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17
Q

Abbe equation

A

r=0.61landa/nsin(alpha)

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18
Q

Magnification in air and oil

A

1000 and 1400

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19
Q

Lighting

A

How light reflects off an object

20
Q

Contrast

A

Difference in appearance between object and its background

21
Q

Bright field microscopy

A

Light microscope
White light passed through specimen
Living sample are nearly invisible
Better image if stained:fixed

22
Q

Bright field stain

A

Stain with dye absorb specific wavelength
Different dye for different biomolecules

23
Q

Phase contrast

A

Light microscope
Use differences in refractive index and thickness
Without the need to section/stain
Turn Phase differences to brightness

24
Q

Differential Interference Contrast

A

Shadow-casting effect
Cells dark on one side
Light on the other side
3D image

25
Q

Fluorescence Microscopy

A

Detect fluorescent proteins
Use laser beam to illuminate a single plane of fluorescently

26
Q

Excitation is ——— wavelength than emission

A

Smaller/shorter

27
Q

Excitation filters

A

Transmit only light of particular wavelengths

28
Q

Dichroic mirror

A

Reflect light below a certain wavelength

29
Q

Emission filter

A

Prevent light from exiting microscope

30
Q

Immunostaining uses ——— to locate specific molecules

A

Antibidies

31
Q

Immunofluorescence microscopy advantage

A

-high specifity
-string signal
-identifies proteins in their native environment

32
Q

Immunofluorescence microscopy disadvantage

A

Experiment on dead cell
Might not be any available antibody

33
Q

GFP

A

Green Fluorescent Protein

34
Q

GFP advantage

A

Cellular events in living cell

35
Q

GFP disadvantaged

A

Introducing the gfp-tagged protein

36
Q

Confocal fluorescence microscopy

A

Optical sectioning: produce in-focus images
3D reconstruction
Distinguish structures in the middle of the cell from top and bottom

37
Q

Advantage of fixed cells

A

1- sectioning thick tissue
2- easier to manage samples
3- no need to recombinant protein

38
Q

Advantage of live cell

A

1- movement of biomolecules within a cell
2- critical context to your observation

39
Q

Electron microscope

A

Use of beam of electrons and electromagnets

40
Q

Electron microscope types

A

1- scanning electron microscopes
2- transmission electron microscope

41
Q

Scan electron microscopes

A

Surface is scanned
Detecting electrons from outer surfaces

42
Q

Transmission electrons microscope

A

Electrons are transmitted through the specimen

43
Q

Cryo-Et microscopy

A

Take multiple 2D images at different angles and crrate a 3D reconstruction

44
Q

Fluorrescense recovery after photobleaching (FRAP)

A

Measure how fast a molecule moves
(Bleach an area and see how long takes for unbleached molecules to return)

45
Q

Fluorescense Resonance Energy Transfer (FRET)

A

Measure if two molecules are touching
(If closer than 10nm one can excite other)

Mbb231 (19 decks)

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