Scanning Microscopy


Although controversy exists concerning the role of chemical fixatives in scanning electron microscopy (SEM) studies of Dipteran larvae, we have observed that filtered 10% formaldehyde solution gives excellent results as a preservative. After immersing in vivo in formaldehyde, the larvae material is preserved for prolonged periods (up to 8 months), before examination with SEM. As a fixative, formaldehyde preserves the structure of the larval cuticle and produces no visible artifacts. Moreover, postfixation is not necessary.

Due to pecularities of the way of life of Wohlfahrtia magnifica (principally the accumulations of necrotic tissue, purulent particles, and other types of substances that often adhere to the numerous spines of larvae), this species must be cleaned before examination by SEM. Manual cleaning with alternating bidistilled water and 0.9% saline solution proved to be a rapid, easy and inexpensive method that gave good results.

Both lyophilization drying and critical point drying were used before sputtering the material. While lyophilization drying proved to be the most effective method for instars II and III, critical point drying was the best technique for study of specimens belonging to instar I. The optimum time for drying and conditions for lyophilization and sputter-coating with gold were determined experimentally. Samples were mounted on SEM stubs with double-sided adhesive and silver conductive paint.

The method proposed is easy and effective for the SEM study of larvae myiasis-producing diptera.

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