Software-Based Stacking Techniques to Enhance Depth of Field and Dynamic Range in Digital Photomicrography

Abstract

Several software solutions are powerful tools to enhance the depth of field and improve focus in digital photomicrography. By these means, the focal depth can be fundamentally optimized so that three-dimensional structures within specimens can be documented with superior quality. Thus, images can be created in light microscopy which will be comparable with scanning electron micrographs. The remaining sharpness will no longer be dependent on the specimen’s vertical dimension or its range in regional thickness. Moreover, any potential lack of definition associated with loss of planarity and unsteadiness in the visual accommodation can be mitigated or eliminated so that the contour sharpness and resolution can be strongly enhanced.

Through the use of complementary software, ultrahigh ranges in brightness and contrast (the so-called high-dynamic range) can be corrected so that the final images will also be free from locally over- or underexposed zones. Furthermore, fine detail in low natural contrast can be visualized in much higher clarity. Fundamental enhancements of the global visual information will result from both techniques.