High-resolution fluorescence images of Drosophila ovaries

Abstract

To compare the differences of caloric density on oogenesis and DNA degradation, adult females were collected at three time points: 0, 2, and 5 days post-eclosion. A total of 81 flies were dissected in the morning to ensure consistency, with no more than 20 minutes between the start of dissection and fixation to prevent tissue degradation. Ovaries were processed following Meehan et al. (2015), staining nuclei with DAPI (Vectashield with DAPI) and detecting DNA degradation with Fluorescein-12-dUTP via the DeadEnd™ Fluorometric TUNEL System (Promega). High-resolution fluorescence images were captured using a BZ-X microscope. Sample sizes of successfully stained ovaries used in the analysis per genetic background, age, and diet can be found in Supplementary Table 2. Oocyte developmental stages were classified according to Jia et al (2016), into five groups: germarium, stages 270 1 – 7, stages 8 – 10, stage 11, and stages 12-14. Images were standardized for size and brightness, converted to black and white using scikit-image v0.22.0 (Walt et al. 2014) and Napari (Chiu et al. 2022) in Jupyter (Kluyver et al. 2016), and annotated via a trained AI model in APEER (Dang et al. 2021). Model training included manual annotation of at least 50 objects per class (developmental stages and background) in a randomly selected images, with iterative updates until accuracy exceeded 95%. The processed image files and the APEER model have been provided in this data repository.