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FluoroFun > 2. & 3. class > Flashcards

2. & 3. class Flashcards

1
Q

What are the dimensions of light & electron microscopy?

A
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2
Q

Epi- and transmission

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3
Q

upright or inverted

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4
Q

upright or inverted

in combination with

transmission or epi

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5
Q

common light sources

general

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6
Q

common light sources

LEDs & lasers

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7
Q

THE QUALITY OF LIGHT

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8
Q

FILTERS

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9
Q

ARRANGEMENT OF FILTERS AND THE EPI-ILLUMINATOR IN THE FLUORESCENCE MICROSCOPE

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10
Q

Fluorescence: enabled color filters

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11
Q

AREA DETECTORS

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12
Q

THE CHARGE-COUPLED DEVICE

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13
Q

area detectors

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14
Q

CCD camera

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15
Q

CCD imagers

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16
Q

CMOS camera

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17
Q

THE PHOTOMULTIPLIER TUBE

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18
Q

THE PHOTOMULTIPLIER TUBE (GaAsP)

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19
Q

avalanche photodiode (APD)

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20
Q

contrast mechanisms

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21
Q

bright field microscope

KOEHLER ILLUMINATION

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22
Q

BRIGHT FIELD MICROSCOPY

staining

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23
Q

DARKFIELD MICROSCOPY

theory

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24
Q

DARKFIELD MICROSCOPY

image interpretation

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25
Q

PHASE CONTRAST

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26
Q

Phase contrast

comparisons

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27
Q

Phase contrast

diffracted wave

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28
Q

Phase contrast

illumination

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29
Q

Phase contrast microscopy

phase plate

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30
Q

phase contrast

positive vs. negative

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31
Q

Polarized light

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32
Q

VECTORIALANALYSIS OF POLARIZED LIGHT USING POLARIZERS

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33
Q

Polarization microscopy

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34
Q

OPTICS OF THE POLARIZING MICROSCOPE

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35
Q

DIC

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36
Q

DIFFERENTIAL INTERFERENCE CONTRAST

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37
Q

Formation of the DIC Image

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38
Q

dic device

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39
Q

phase contrast and dic pictures

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40
Q

DIFFRACTION

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41
Q

INTERFERENCE

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42
Q

THE DIFFRACTION IMAGE OF A POINT SOURCE OF LIGHT

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43
Q

DIFFRACTION AND SPATIAL RESOLUTION: NUMERICAL APERTURE (NA)

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44
Q

REFRACTION:

A
45
Q

THE CONSTANCY OF OPTICAL PATH LENGTH BETWEEN OBJECT AND IMAGE

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46
Q

SPATIAL RESOLUTION

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47
Q

Ryleigh criterion

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48
Q

Resolution:

typical values

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49
Q

DEPTH OF FIELD AND DEPTH OF FOCUS

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50
Q

Why Fluorescence?

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51
Q

Background free!

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52
Q

Compare absorption and fluorescence!

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Fluor is like stars in the sky!

53
Q

Epi-Fluorescence-Microscopy

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54
Q

In the most simple form of epi-fluorescence microscopy

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55
Q

Nyquist criterion!

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56
Q

CONFOCAL FLUORESCENCE MICROSCOPY

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57
Q

laser scanning confocal

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58
Q

Imaging speed in confocal microscopy

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59
Q

wide field vs confocal

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60
Q

In confocal microscopy, the pinhole

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61
Q

TIRF MICROSCOPY

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62
Q

TIRF: critical angle?

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63
Q

A (commercial) TIRF module

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64
Q

TIRF:

resolution? contrast?

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65
Q

TWO-PHOTON EXCITATION FLUORESCENCE MICROSCOPY

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66
Q

TWO PHOTON EXCITATION:

pulsed laser

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67
Q

TWO PHOTON:
rayleigh

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68
Q

TWO PHOTON:
image

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69
Q

TWO PHOTON:

Light scattering and absorption processes

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70
Q

Summary

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71
Q

FAST (REAL-TIME) IMAGING IN CONFOCAL MICROSCOPY

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72
Q

Tandem scanning confocal microscopy using a spinning Nipkow disk

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73
Q

Selective plane illumination

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74
Q

Light-sheet microscopy features

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75
Q

Different Light Sheet Fluorescence Microscopy configurations

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76
Q

Selective plane illumination:

The fundamental principle is the detection of fluorescence light

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77
Q

dichromatic mirror

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78
Q

cross talk

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79
Q

cross talk

problem & solution

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80
Q

cross talk

sequential imageing

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81
Q

spectral imaging

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the pixel intensity versus the center wavelength of each emission band. The accuracy of the emission spectra obtained by this technique depends largely on the number of images gathered at distinct wavelength bandwidths, the bandwidth size (shorter bandwidths yield more accurate spectra), specimen quality, and the instrument detector sensitivity. In confocal microscopes that use separate pinholes for each detector, wavelength-dependent variations can occur in optical section thickness for the different spectral channels. However, most modern instruments are designed with a single pinhole for all detectors and, in any event, probes with highly overlapping spectra are usually very similar in spectral range.

82
Q

linear unmixing

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83
Q

Fluorescence spectrum detection

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84
Q

Airy scan

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85
Q

Airy scan
Improving the performance of a confocal laser scanning microscope

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86
Q

Summary of resolutions

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FluoroFun (9 decks)

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