Drosophila Pupal Wing Imaging Protocol

To prepare the Drosophila pupal wing samples for image collection, pupae at appropriate ages were fixed to double-sided tape and the pupal case above the left wing was removed. The pupae were then placed on a small drop of water or Immersol W 2010 (Zeiss 444969-0000-000) in a glass bottom dish with the left side facing downward^{6 (link),7 (link),46 (link),66 (link)}. The fixed time-point images other than Fig. 3a, b and time-lapse images shown in Fig. 6l, m were acquired using an inverted confocal microscope (A1R; Nikon) equipped with a ×60/NA1.2 Plan Apochromat water-immersion objective at 25 °C. Other images were acquired using an inverted confocal spinning disk microscope (Olympus IX83 combined with Yokogawa CSU-W1) equipped with an iXon3 888 EMCCD camera (Andor), an Olympus ×60/NA1.2 SplanApo water-immersion objective, and a temperature control chamber (TOKAI HIT), using IQ 2.9.1 (Andor)^{66 (link)}. After imaging, we confirmed that the pupae survived to at least the pharate stage.

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Ikawa K, & Sugimura K. (2018). AIP1 and cofilin ensure a resistance to tissue tension and promote directional cell rearrangement. Nature Communications, 9, 3295.

Publication 2018

Immersol W 2010 CSU-W1 iXon3 888 EMCCD camera

Confocal microscope Dish Drosophila Immersion Pupae Samples collection

Corresponding Organization :

Other organizations : Kyoto University

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Variable analysis

independent variables

Age of Drosophila pupae

dependent variables

Pupal wing morphology

control variables

Fixation of pupae to double-sided tape
Removal of pupal case above the left wing
Positioning of pupae with left side facing downward on a drop of water or Immersol W 2010
Temperature of imaging setup (25 °C or controlled chamber)
Imaging equipment (inverted confocal microscopes)

positive controls

Confirmation of pupal survival to at least the pharate stage after imaging

negative controls

No negative controls were explicitly mentioned in the input protocol.

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