Scientific Volume Imaging B.V. Nyquist rate and PSF calculator Nyquist calculator


This is the parameter list used in this calculation:

Parameter Value
Microscope type
Numerical aperture
Excitation wavelength
Emission wavelength
Number of excitation photons
Lens immersion refractive index
Backprojected pinhole radius
STED Depletion wavelength
STED 3x percentage
STED Saturation factor
Specimen medium refractive index
Specimen medium refractive index
SPIM excitation mode
Width of Gaussian light sheet
Light sheet NA
Light sheet fill factor
Backprojected pinhole radius
B.P. distance between pinholes

The optical axis lays along z. Your Nyquist sampling is:

x: nm
y: nm
z: nm
Set your zooms and scanning steps so that each pixel covers a x-y area of nm × nm (or smaller)
Calibrate and set your z-stepper so that it takes steps of nm when acquiring a 3D stack (or smaller)

PSF: XY MIP grayscale
xy MIP grayscale
PSF: XY MIP false color
xy MIP false color

PSF: XZ MIP grayscale
xz MIP grayscale
PSF: XZ MIP false color
xz MIP false color

These are xy and xz MIP projections of the 3D point spread function.
The 3D image dimensions (x × y × z) are × × µm.
It was calculated using the Nyquist sample sizes specified above.
The number of samples (x × y × z) to cover this volume at the Nyquist rate is × × .
To enhance dimmer regions the gamma is corrected by 1.7 in the false color image.

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