Comparison of protein A-gold and ferritin immunoelectron microscopy of Semliki Forest virus in mouse brain using a rapid processing technique.

Abstract

Processing tissue for transmission electron microscopy by standard laboratory methods can take two to three days. This makes the development of new techniques time consuming and generally restricts the use of the electron microscope in routine diagnostic work. The possibility of viewing tissue with the electron microscope five hours after sampling using rapid processing techniques is presented. The morphology of the tissue appears undamaged with cell and organelle ultrastructures being readily recog

Applications of immunogold labelling in animal and plant virology

“…Excessive damage due to freezing may be avoided by first soaking the tissue in a cryoproteetant such as 10% DMSO [12]. There is one report, describing immunogold labelling of cryostat sections of mouse brain infected with Semliki Forest virus, in which the tissue was fixed in 1% glutaraldehyde [23]. This may indicate that the fixation protocol required to achieve reasonable penetration is not identical tbr different tissues.…”

“…This may indicate that the fixation protocol required to achieve reasonable penetration is not identical tbr different tissues. Although immunogold reagents have been successfully used in the pre-embedding procedures to detect virus antigens in cell monolayers [73] and tissue [23], there are some disadvantages to the technique. The permeabilization and freezing steps usually result in some loss of detailed morphology.…”