Calculating the pixel sizes on images

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Great post and great blog on all things relating to fluorescent microscopy.

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In most microscopy images that are published in research papers, there appears a scale bar.  The scale bar is like a ruler that allows you to compare sizes and distances in images from different sources.  Although a scale bar is helpful for assessing by eye, many image processing programs allows you to measure distances in the image. The problem is that these measurements are in pixels. That is what I encountered when I wanted to measure certain objects in my images. How to convert from pixels to nanometers (or microns) requires a simple formula and some prior data as follows:

  1. Objective magnification
  2. Lens magnification (in some microscopes, it is possible to get extra mag of 1.25x, 1.6x or 2x.
  3. C mount (is usually 1x)
  4. Pixel size – is the actual pixel size of the camera that is attached to the microscope.
  5. Binning – i.e. combining a cluster of pixels to a…

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