Object Size & Pixel Size
Object Size & Pixel Size
Do not forget, it's A Matter of Scale
Rule of Thumb
Example
Limit
If your object size is below the limit of diffraction (see Abbe diffraction limit) , i.e. roughly 250 nm, there is no need to acquire images with pixel size of 2 Å !
Please use this Nyquist calculator to obtain the smallest meaningful pixel (voxel) size that matches your microscope's capabilities.
If needed, you can lower the obtained value by 1.5 (in XY) and still obtain high quality images.
Object size and intensity measurement
With an inadequate pixel size, the measured size and intensity of your object will be bias.
In the example below, you can see that a translation of only half a pixel would make the object twice as large and twice as dim in the case of an inadequate pixel size (left), and be without consequence with optimal sampling (right)
Range
What is true for XY, is also true for Z.
Please do not make z-stack acquisition, with a pixel size of 40nm and a z-step of 2um!
- Last Author
- romainGuiet
- Last Edited
- Dec 7 2018, 11:09