Field Emission Scanning Electron Microscopic (FESEM) Analysis of Treated Larvae
This protocol is extracted from research article:
Bioefficacy of Lemongrass and Tea Tree Essential Oils Against House Fly, Musca domestica
Proc Natl Acad Sci India Sect B Biol Sci, Feb 18, 2021; DOI: 10.1007/s40011-020-01220-z

To study the effect of EOs on insect morphology, 10 larvae were exposed to LC50 concentrations for 24 h. After treatment, the larvae were prepared for FESEM analysis to observe the changes in integument and surface structures. The morphology of the treated and control larvae were observed in an electronic microscope (FESEM 450, FEI Nova Nano, USA) complemented with EDS system at a voltage of 3 kV. For FESEM visualization, the samples were primarily fixed in 2.5% glutaraldehyde for 12 h, then washed with distilled water for 20 min followed by a secondary fixation with 4% Osmium tetroxide. The samples were further dehydrated with different ethanolic grades (25, 50, 75, 90 and 100% ethanol) each for 10 min at RT. Finally, the samples were subjected to critical point drying (CPD) and mounted on carbon tape followed by sputter coating with 100 Å gold.

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