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I am trying to reconstruct SIM images taken from Lattice light-sheet microscope. It has only one directional image and is in .tif format. I am struggling to understand the following things while running the software. Can someone kindly help me to reconstruct SIM image
To create OTF
- How should my input PSF be? I mean, should it be a 3D PSF stack (XYZ) without phases or should it be a 3D PSF stack of (XY, Phase, Z)?
About the image and parameters - As I understand, if I have a 3D images (XYZ) (suppose 50 z planes) where each plane is imaged 5 times for different phases (means different X-galvo positions), I can arrange my images either in a single stack in XY,phase,Z (total 50*5 = 250 number of sclices as shown in the repository data) or 50 number of XY,phase stacks with similar file name pattern. Am I correct?
- Since I have only one angle should I mention 0 against k0angle argument?
- If I have a lattice line spacing of 1 micron and if I take 5 phases in between this 1 micron, what would be the correct value of ls (1 or 0.2)?
- If I organize my images the way I mentioned in point 1 what would be the right value of fastSI?
- If my illumination objective’s NA is 0.8, and the outer and inner NA of my ring are 0.7 and 0.6 respectively what would be the right value of besselNA?
I appreciate your support.
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