2. & 3. class Flashcards
(86 cards)
1
Q
What are the dimensions of light & electron microscopy?
A
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
A
25
PHASE CONTRAST
26
Phase contrast
comparisons
27
Phase contrast
diffracted wave
28
Phase contrast
illumination
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Phase contrast microscopy
phase plate
30
phase contrast
positive vs. negative
31
Polarized light
32
VECTORIALANALYSIS OF POLARIZED LIGHT USING POLARIZERS
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Polarization microscopy
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OPTICS OF THE POLARIZING MICROSCOPE
35
DIC
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DIFFERENTIAL INTERFERENCE CONTRAST
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Formation of the DIC Image
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dic device
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phase contrast and dic pictures
40
DIFFRACTION
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INTERFERENCE
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THE DIFFRACTION IMAGE OF A POINT SOURCE OF LIGHT
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DIFFRACTION AND SPATIAL RESOLUTION: NUMERICAL APERTURE (NA)
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REFRACTION:
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THE CONSTANCY OF OPTICAL PATH LENGTH BETWEEN OBJECT AND IMAGE
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SPATIAL RESOLUTION
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Ryleigh criterion
48
Resolution:
typical values
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DEPTH OF FIELD AND DEPTH OF FOCUS
50
Why Fluorescence?
51
Background free!
52
Compare absorption and fluorescence!
Fluor is like stars in the sky!
53
Epi-Fluorescence-Microscopy
54
In the most simple form of epi-fluorescence microscopy
55
Nyquist criterion!
56
CONFOCAL FLUORESCENCE MICROSCOPY
57
laser scanning confocal
58
Imaging speed in confocal microscopy
59
wide field vs confocal
60
In confocal microscopy, the pinhole
61
TIRF MICROSCOPY
62
TIRF: critical angle?
63
A (commercial) TIRF module
64
TIRF:
resolution? contrast?
65
TWO-PHOTON EXCITATION FLUORESCENCE MICROSCOPY
66
TWO PHOTON EXCITATION:
pulsed laser
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TWO PHOTON:
rayleigh
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TWO PHOTON:
image
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TWO PHOTON:
Light scattering and absorption processes
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Summary
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FAST (REAL-TIME) IMAGING IN CONFOCAL MICROSCOPY
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Tandem scanning confocal microscopy using a spinning Nipkow disk
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Selective plane illumination
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Light-sheet microscopy features
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Different Light Sheet Fluorescence Microscopy configurations
76
Selective plane illumination:
The fundamental principle is the detection of fluorescence light
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dichromatic mirror
78
cross talk
79
cross talk
problem & solution
80
cross talk
sequential imageing
81
spectral imaging
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
linear unmixing
83
Fluorescence spectrum detection
84
Airy scan
85
Airy scan
Improving the performance of a confocal laser scanning microscope
86
Summary of resolutions
