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This is a confocal microscopy class. So please answer clear and detail according to the subject. Thank you

1.Explain how light diffraction causes the lateral point spread function of light as shown by Airy. Illustrate with a diagram of the PSF in the lateral direction.

2.What is the relative size of the point spread function in the vertical compared to lateral direction and how does this affect the shape of the voxels we record in a optimal resolution z-stack.

3.Is the point spread function wider or narrower with large aperture lenses? Explain why.

4.Does a larger aperture lens have higher resolution (more detail) or lower resolution (poor detail)? Or is aperture size unrelated to resolution?

5.Definedynamicrange.Explaintheimportanceofadjustingthedynamicrangeof the light intensity sensors (PMTs) and how the confocal encodes intensity into numbers.

6.Explain the advantage and disadvantage of frame-averaging.

6b.Explain how to optimize the resolution of your scanned image to avoid oversampling and minimize file size.

7.Make a list of the most functionally-important parts inside of the scan head box and briefly explain what each part does. Hint1: there are 5 kinds of things, and the AOTFs & lasers are not in the scan head box. Hint2: Without these 5 things, it would not be a laser scanning confocal microscope.

8.Explain why immersion oil is used for some lenses. Explain how it works.

9.Explain the relationship between the point-spread function and super-resolution.

10. What is chromatic aberration?

11. How does one detect cross-talk and bleed-through? How can you avoid it, and how can you correct for it if you could not avoid it?

12. Describe the three main ways to manage autofluorescence.

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Patrina Schowalter
Patrina SchowalterLv2
28 Sep 2019

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