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All the single molecule fluorescence experiments were performed with a time-resolved confocal fluorescence scope fitted with picosecond lasers (465 nm and 636 nm at 20 MHz) and optics for single molecule fluorescence spectroscopy and lifetime imaging (Microtime 200, Picoquant, Berlin, Germany).
After another 24 h, the cover slips were taken and placed on the fluorescence scope (the cover slips actually form the bottom of the flow chamber).
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GFP expression in whole larvae or adults was documented under a fluorescence dissecting scope with a GFP filter.
Fat bodies were carefully dissected, fixed and stained with Nile Red and documented under a fluorescence dissecting scope using a rhodamine filter.
All the F0 strains were balanced by TM6B, Dfd:YFP, Tb or CyO Dfd:YFP, and the non-YFP embryos were collected using a fluorescence dissection scope (Zeiss).
For bright GFP constructs, it should be possible to screen using a fluorescence dissecting scope.
Expression of GFP-tagged protein was monitored using a fluorescence dissection scope.
Fish were gently submerged in a Petri dish containing 0.005% MS-222 and mounted on a Leica fluorescence dissecting scope with a FITC filter set (Leica Microsystems Inc., Bannockburn, IL).
Progeny were dissected in PBS, and brains were immediately inspected for GFP expression using a fluorescence dissection scope (Olympus, model BX51W1).
Larvae were observed under a fluorescence dissecting scope, and we established an arbitrary area of approximately five cell diameters above and below the horizontal myoseptum, which runs from the first somite to the end of the tail.
Transgenic animals were scored under the fluorescence-dissecting scope using co-injection marker p unc-122 GFP. unc-122 GFP
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