Reflection contrast microscopy performed on epi‐illumination microscope stands: Comparison of reflection contrast‐ and epi‐polarization microscopy

Abstract

SUMMARYIn reflection contrast microscopy (RCM) special optics such as a quarter‐wave plate and a central stop are used, the latter being unique for Leitz microscope stands and contributing much to contrast enhancement of the image. In this paper we demonstrate that a cap or prism, linked by oil immersion to the glass slide, can be used instead of this central stop. Thus, microscope stands with incident illumination may be used for RCM.We systematically investigated RCM and epi‐polarization microscopy (EPM), both incident light microscopical methods, for visualization of immunogold‐silver‐stained cells, autoradiographic stained cells and immunoperoxidase‐diaminobenzidine stained chromosomes. RCM was found to be superior (1) in terms of the intensity of reflected light from reflecting objects, (2) in the visualization of unstained structures and background and (3) with respect to the spatial resolution that is achieved, especially in the tiny stained structures in the image.