Larval Fillet Preparation: A Method to Visualize Intact Sensory Neurons and Associated Epidermal Cells

Abstract

Source: Tenenbaum, C. M., Gavis, E. R. Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells. J. Vis. Exp. (2016).

This video describes how to prepare larval fillets used to visualize sensory dendritic arborization (da) neurons and their associated epidermal cells. The featured protocol shows in detail how to remove muscle tissue from the body wall while preserving the cells' morphology, which improves immunostaining and visualization of the otherwise obscured da neurons and epidermal cells.

Protocol

This protocol is an excerpt from Tenenbaum and Gavis, Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells, J. Vis. Exp. (2016). NOTE: The procedure for muscle removal (Figure 1) is a modification of previously described methods for preparing larval fillets. The steps that precede and follow muscle removal are…

Representative Results

Figure 1: Overview of Muscle Removal Procedure. A larva is dissected and pinned to a silicone elastomer dissecting dish. To remove muscle tissue, one forceps prong is inserted between the muscle and epidermal cell layer, starting from the dorsal midline near a segment boundary (2.2). The forceps is pulled upwards to break the attachment between the muscle and body wall (2.3). This is repeated for segments of interest…

Materials

Dumont #5 tweezers	Electron Microscopy Sciences	72701-D
Micro Scissors, 8 cm, straight, 5 mm blades, 0.1 mm tips	World Precision Instruments	14003
Sylgard 184 silicone elastomer kit	Dow Corning	3097358-1004	for dissecting plates
Austerlitz insect pins, 0.1 mm	Fine Science Tools	26002-10
Fostec 8375 light source	Artisan Technology Group	62792-4