Advantage image capture software

Tissues for electron microscopic examination were prepared as described in⁴² . Thoraces were fixed with 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1 M sodium cacodylate buffer, pH7.4, overnight at 4 °C. After subsequent buffer washes, the samples were post-fixed in 2.0% osmium tetroxide for 1 h at room temperature, and then washed again in buffer followed by dH₂O. After dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Thin sections were stained with lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.

Heads from female Repo > UAS-20xShi¹ flies approximately two weeks post-eclosion were dissected in a cold room in PBS and fixed with 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1M sodium cacodylate buffer, pH7.4, overnight at 4°C. Samples were post-fixed in 2.0% osmium tetroxide for 1 hour at room temperature, and rinsed in DH₂O prior to en bloc staining with 2% uranyl acetate. After dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Flies were not strictly circadian entrained or taken at an exact time of day, but dissections were performed together in the afternoon. Reported images represent sections from single flies from each genotype, although the experimental was compared to both parental controls. The described microtubule structures were not seen in the controls. Embedding, staining and sectioning was performed by the Electron Microscopy Resource Laboratory at Penn. Thin sections were stained with uranyl acetate and lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.