SCREENING METHODS: Confocal laser scanning microscopy(part 1)

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After all these treatment finally the stained sections will be observed with a transmission electron microscope. Ultrathin sections obtained by conventional procedures, namely fixation with aldehydes, post-fixation with osmium tetraoxide, dehydratation and embedding in Epoxy resin, allow the observation of membrane and cytoplasmatic alterations. Treatment with AMPPs can induce several external and internal changes such as membrane bleb, ruffling or detachment, the presence of electrodense dots or fibers, hypodense cytoplasmic release and cell vacuolization.

Most importantly From the TEM micrograph we can observe the changes caused by the plant extract on fungal cytoplasm.

Although using SEM and TEM a detail ultrastracture can be obtained, it is also necessary to monitor the overall integrity and viability of the fungal cells after exposure to antimicrobial compounds. Using confocal microscopy, the kinetics and morphological evolution of microbial cell population can be monitored at real time and compared with microorganism viability.

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