Image Processing Of Muscle Striations Below The Resolution Limit Of The Light Microscope

We describe the use of digital deconvolution in the study of muscle striations below the resolution imposed by the optical diffraction of our video light microscope. To use deconvolution procedures on muscle images, the transfer function of the optical system is first characterized. This is accomplished by imaging a step object and fitting the image with the combination of a first order Bessel and Gaussian function using a non-linear least squares approach. Due to the ill-conditioned nature of deconvolution, however, ambiguity in the reconstruction is sometimes found. To allow better estimation of the true object, separate deconvolution approaches are used and the reconstructions compared. In this manner, the fine structure of muscle striations is determined.