Resolution - Answer-how close two objects can be brought together and still be
visualized as two distinct objects
Smaller resolution value = - Answer-better resolution
What does resolution depend on? - Answer-- size of the object
- contrast between the two objects
Airy disc - Answer-- Light passing through an aperture (ex. objective) is diffracted
- Light from single point will create pattern of rings
(image is actually a diffraction pattern of an object)
What part of airy pattern has the highest portion (84%) of the light intensity? - Answer-
The central part (0 order)
- light intensity decreases with increasing order number
0 peak = 84%; most intensity
100% = zero order peak + 1 order + 2 order....
Point spread function (PSF) - Answer-Spread of intensity due to microscope; should be
symmetrical
- If not well aligned, microscope can't reach optimum resolution
Rayleigh criterion - Answer-2 points are "resolved":
- Minimum of the 0 order peak of the 1st airy disk equals the maximum of the 0 order
peak of the 2nd airy disk
theoretical dxy = - Answer-full width at half max of zero order peak
(resolution depends on how broad this zero order peak is)
1 airy unit = - Answer-diameter of zero order peak
(this is considered the best confocal resolution)
An optical confocal aperture will capture.... - Answer-all of the zero order peak
What happens if aperture is too small? - Answer-Blocks the zero order peak
What happens if aperture is too large? - Answer-Will collect out of focus light
Optical resolution - - Answer-minimum distance that two objects must be separated
Resolution is determined by the diameter of the.... - Answer-airy disc
Resolution is what-limited? - Answer-Diffraction limited