Modified thiocarbohydrazide procedure for scanning electron microscopy: routine use for normal, pathological, or experimental tissues

Abstract

A modified thiocarbohydrazide (TCH) technique to cross-link osmium layers has resulted in a reliable method for preparing a wide variety of soft biological tissues for scanning electron microscopy without the use of evaported metal. The technique works equally well on tissues with smooth surfaces and those with abrupt changes in contour or cut surfaces, as in biopsy or autopsied material or pathologically altered tissues, and thus has wide applicability. Small surface structures and junctional areas between cells are distinct. In addition, thin cells such as alveolar endothelium in the lung exhibit a transparent property, allowing the visualization of cells within the capillary lumen while retaining adequate contrast for study of the capillary wall itself. Absence of an evaporated metal coat makes the removal of tissue from the scanning electron microscope specimen easy for embedding and examination of the same material with the transmission electron microscope.