The preparation of human articular cartilage for scanning electron microscopy

This paper compares sixteen preparative techniques thought to be of advantage in the study by scanning electron microscopy (SEM) of human articular cartilage surfaces. The adequacy of surface preservation obtained with the techniques, was judged subjectively, first, by the reproducubility of secondary electron images of normal cartilage, and second, by comparing the results with those obtained by reflected light microscopy of the fresh unfixed cartilage surface over a magnification range of X20- X240. Adequate surface preservation was confirmed when cartilage surfaces had been dehydrated through ethanol to propylene oxide and vacuum dried; dehydrated through amyl acetate and quenched in Freon before freeze-drying; dehydrated and passed through amyl acetate at low temperature before freeze drying. Valuable information can be obtained from different specimens by varying the technique of preparation. At different ages, different surface features are best preserved. In a systematic study it has been found essential to adopt a uniform preparative method and to control the results by reflected light microscopy. Even with the most perfect preparation, the surface appearances cannot be identical with those that function under load in vivo.