Scanning electron microscopy of cell surfaces following removal of extracellular material

Abstract

The application of scanning electron microscopy to the study of cell surfaces is limited in intact tissues, because extracellular material may often obscure the details of nonluminal surfaces. To remove connective tissue elements we have treated human skin and both kidney, and an autonomic ganglion of the rat with hydrochloric acid and collagenase. Regional variations in the basal surface of the nephron are noted following removal of the basement membrane. The basilar interdigitations of the cells of the proximal tubule appeared as parallel ridges encircling the tubule. Ridges on the parietal epithelium of Bowman's capsule were randomly arranged and alternated with smooth surfaces. The dermal surface of the human epidermis has an alveolar or honeycomb appearance due to the elevation of the epidermal ridges and numerous pits for the dermal pegs. At higher magnifications the basal surface of cells of the stratum germinativum possessed numerous and irregular projections. Neurons with their processes are evident in the autonomic ganglion. The soma of the neurons are enclosed by flattened satellite cells. Irregular spaces between opposed satellite cells are interpreted as regions for the passage of processes related to the ganglion cells. Nodes of Ranvier were clearly seen along nerve fibers. Some pitting of the nerve fibers was also noted. The HCl-collagenase method has the advantage of the removal of collagen and basement membrane while preserving the structural integrity of the cell surface.

Renal corpuscle of the sturgeon kidney: An ultrastructural, chemical dissection, and lectin‐binding study

“…Other samples were first subjected to chemical and mechanical microdissection. For chemical microdissection, the specimens were digested either with HCl (Evan et al, 1976) or KOH (Ushiki and Murakumo, 1991). In both cases, collagenase digestion was omitted.…”

Surface View of Pericytes on the Retinal Capillary in Rabbits Revealed by Scanning Electron Microscopy

“…A new technique, introduced by EVAN et al (1976) extracellular materials with HCl and collagenase, made it possible to visualize the three dimensional structure of the cell surface.…”

Scanning electron microscopic observations of rat tibia using the HCl-collagenase method.

“…The metaphysis of rat tibia was examined by scanning electron microscopy (SEM) using the HC1-collagenase method (EVAN et al, 1976) and the three-dimensional structure of osteocytes and the relationship between osteocytes and osteoblasts were demonstrated.After the HC1-collagenase treatment, the bone matrix near the exposed surface was completely removed but the cellular components retained their structural integrity.Young osteocytes located close to osteoblasts showed flattened, spindly shapes. They were connected with adjacent osteoblasts by short, wide and slightly branched processes.…”

“…The metaphysis of rat tibia was examined by scanning electron microscopy (SEM) using the HC1-collagenase method (EVAN et al, 1976) and the three-dimensional structure of osteocytes and the relationship between osteocytes and osteoblasts were demonstrated.…”

“…The present study was designed to reveal the three-dimensional structure of osteocytes and the relationship between osteocytes and osteoblasts or among osteocytes after removing the bone matrix using the HCl-collagenase method originated by EVAN et al (1976). …”