Ovaries from 2- to 3-day-old yeasted females were dissected and fixed with cacodylate/formaldehyde as described by Hughes et al. (2011) (link). For embryos, mated females laid on grape plates for 2 h, and then embryos were fixed in heptane/methanol as described by Bonner et al. (2013) (link). Rat anti-α-tublin primary antibody (1:250, BioRad) was used with Alexa-488 or Alexa-555 conjugated secondary antibody (Molecular Probes, 1:350). DNA was then labeled with DAPI (2 μg/ml), and samples were mounted in ProLong Gold (Invitrogen).
For all imaging, the DeltaVision microscopy system (Applied Precision), equipped with an inverted Olympus 1670 microscope and a high-resolution CCD camera, was used. All acquired images were then deconvolved using SoftWoRx software (Applied Precision).
For all imaging, the DeltaVision microscopy system (Applied Precision), equipped with an inverted Olympus 1670 microscope and a high-resolution CCD camera, was used. All acquired images were then deconvolved using SoftWoRx software (Applied Precision).
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