High voltage electron microscopy of biological materials

Serial block face scanning electron microscopy and the reconstruction of plant cell membrane systems

“…The zinc iodide technique is suitable for the selective staining of higher plant, algal and fungal tissues and has been combined with thick sectioning in a number of studies on the endomembrane system (e.g. Harris, 1979;Hawes, 1981;Hawes et al, 1981). This technique can be used for the impregnation of nuclear envelopes, endoplasmic reticulum (ER; Figs.…”

“…They rely on the reduction of osmium tetroxide on the surface of membranes or in cisternal spaces. The reduction of osmium tetroxide is chemically aided by potassium ferrocyanide (Hepler, ) or zinc iodide (Hawes, ; Hawes, ; Hawes et al ., ). The zinc iodide technique is suitable for the selective staining of higher plant, algal and fungal tissues and has been combined with thick sectioning in a number of studies on the endomembrane system (e.g.…”

“…If three‐dimensional (3D) or volume information on cell and tissue structure is needed, then extremely tedious serial section and 3D reconstruction techniques are required (Bang & Bang, ). In an attempt to overcome this problem, in the late 1960s and 1970s, high‐voltage electron microscopes (1–3 MeV) were used to image either extremely thick 1 μm plus sections (Glauert, ; Hawes, ; Hawes et al ., ) or whole freeze‐ or critically point dried cells (Porter & Anderson, ; Hawes, ), which were subsequently analysed using stereoscopic viewing and measuring techniques. More recently, electron tomography of semithick resin sections has become the technique of choice for high‐resolution 3D imaging of organelles (Austin & Staehelin, ).…”