An investigation of the use of chromium, platinum and gold coating for scanning electron microscopy of casts of lymphoid tissues.

Resin casts replicate the internal structure of organs and provide a three-dimensional representation of the arrangement of vessels and intercellular spaces. Casting media are insulators and must be coated with a conductor to prevent sample charging and to allow the adequate production of secondary electrons from the specimen to generate sufficient signal to form a clear image. Visualization of surface structures depends largely on the metal coating. The use of gold or platinum, deposited on Mercox casts of lymphoid tissues using plasma-magnetron sputtering, and of chromium coating of casts by Penning ion-beam coating, was investigated. Casts were examined using a field emission scanning electron microscope at 3-3.5 kV. Thick coatings of gold were necessary to reduce cast charging but they obscured fine structural information. Charging effects were less pronounced when casts were coated with platinum, but charge lines were present at slow scan rates. The dimensions of cast impressions for both platinum and chromium coatings were similar to those described in fixed tissues. Negligible charging and maximal cast thermal stability and structural information was obtained from casts which were tumbled during chromium coating.