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Mounting method for live-imaging of Drosophila pupa

This protocol allows for live imaging of intact pupae.

Figure 1. Stick the intact pupa (ventral side down) to a microscope slide coated with double-sided tape

Figure 1. Stick the intact pupa (ventral side down) to a microscope slide coated with double-sided tape

drosophila_pupa_mounting_protocol_Fig2

Figure 2. Under the dissection microscope, gently remove the pupal case and carefully pick the pupa with a brush

drosophila_pupa_mounting_protocol_Fig3

Figure 3. Place the spacer on the glass surface of the CherryTemp chip (engravements facing down, coverslip and liquid microfluidic chamber facing up, in contact with the spacer)

drosophila_pupa_mounting_protocol_Fig4

Figure 4. Gently place the pupa on the glass surface of the CherryTemp chip. Place it with the desired orientation (imaging zone facing up). Important: place the pupa centered in the chamber, which corresponds to the thermalization zone

drosophila_pupa_mounting_protocol_Fig6

Figure 5. Place a high resolution glass coverslip on top of the mounted sample. The surface will be flat and the chamber sealed.

** For an inverted microscope configuration flip the mounted system  180°C

Mounting protocol adapted for the Cherrytemp technology from:

  • Our user’s experience
  • Zitserman,D. and Roegiers, F. J Vis Exp 2011; (51): 2706

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