Low Temperature Scanning Electron Microscopy of Soft Bodied Organisms

Abstract Resolving the true fine structure and behavior of soft bodied organisms such as arthropod larvae, acari, nematodes, annelids and pupae by conventional scanning electron microscopy has been difficult. Classical preparation techniques, involving immersion fixation, dehydration and critical point drying, cause significant deleterious changes in specimen structure due to effects of surface tension, osmotic stress and mechanical damage. The intact exoskeleton, cuticle or other limiting membranes of these organisms, creates a formidable obstacle to the penetration of fixatives and solvents, which are only effective after a long period of time (hours). During this time, behavioral postures and much of the structural integrity are lost and most organisms detach from their hosts. In previous studies, our laboratory utilized methods in low temperature freeze stabilization, to preserve and observe the undisturbed fine structure of biological samples. Plunge freezing, although more rapid, resulted in many parasitic organisms dislodging from hosts and being lost.