Detecting Pole Cells in Drosophila Embryos

Pole cells were detected in fixed 10- to 14-hour-old embryos using anti-Vas immunohistochemistry as previously described [60 (link)]. The following antibodies were used: 1:500 rabbit anti-Vas and 1:500 biotin goat anti-rabbit (Jackson Immuno Research Laboratories). Embryos were mounted in 80% glycerol and analyzed using a Zeiss compound microscope. Images of representative embryos were captured on a Nikon DsQi-2 microscope using a 20 × 0.75 NA air objective and DIC optics.

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Hakes A.C, & Gavis E.R. (2023). Plasticity of Drosophila germ granules during germ cell development. PLOS Biology, 21(4), e3002069.

Publication 2023

compound microscope DsQi-2 microscope

Antibodies Biotin Compound microscope Embryos Glycerol Goat Immunohistochemistry Microscope Optics Pole cells Rabbit

Corresponding Organization : Princeton University

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Variable analysis

independent variables

Age of embryos (10- to 14-hour-old)

dependent variables

Presence of pole cells detected using anti-Vas immunohistochemistry

control variables

Method of fixation and mounting of embryos
Microscope and imaging setup (Zeiss compound microscope, Nikon DsQi-2 microscope, 20 × 0.75 NA air objective, DIC optics)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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