Objective evaluation of the relative modulation transfer function of densitometers for digitisation of electron micrographs

Digitisation of images recorded on film is a crucial part of data acquisition in electron microscopy, particularly for electron cryo-microscopy of biological specimens where the contrast and signal-to-noise ratio are low. A quantitative method to evaluate and compare the quality of densitometers, as measured by the modulation transfer function (MTF), is described here. The densitometer is modelled as a linear system, the output being the convolution of the input image and a point spread function. The MTF is the magnitude of the Fourier transform of the point spread function. The relative MTF describes the quality of signal transfer with spatial frequency. It is important that fine structural details in the micrograph are digitised with a high value for the MTF which does not vary with direction. A test pattern has been generated by projecting an electron image of a grid pattern onto film. The film is scanned and a computer program measures the intensities of the diffraction orders of the repeating pattern. Three different scanners are compared, one is a point scanner and the other two are line scanners. The test can be used to check if a scanner is set up optimally, and how it compares with another scanner.