Macromolecular structures of biological specimens are not obscured by controlled osmium impregnation

High magnification imaging of macromolecules on metal coated biological specimens is limited only by wet preparation procedures since recently obtained instrumental resolution allows visualization of topographic structures as smal l as 1-2 nm. Details of such dimensions may be visualized if continuous metal films with a thickness of 2 nm or less are applied. Such thin films give sufficient contrast in TEM as well as in SEM (SE-I image mode). The requisite increase in electrical conductivity for SEM of biological specimens is achieved through the use of ligand mediated wet osmiuum impregnation of the specimen before critical point (CP) drying. A commonly used ligand is thiocarbohvdrazide (TCH), first introduced to TEM for en block staining of lipids and glvcomacromolecules with osmium black. Now TCH is also used for SEM. However, after ligand mediated osinification nonspecific osmium black precipitates were often found obscuring surface details with large diffuse aggregates or with dense particular deposits, 2-20 nm in size. Thus, only low magnification work was considered possible after TCH appl ication.