Picture from Gupta N et al. (2024) MicroPubl Biol "A DTC morphometrics package for quantification of complex and variable cellular ...."

Figure 1. Overview of image processing and markup.: A) A maximum projection image of an Z-confocal series (file format .nd2), with no contrast adjustment or filtering. B) Maximum projection image created from the same image file, using "Max project and change contrast" command. C) Same panel as in B, with yellow segmented line showing path used in "Rotate and straighten based on curved line" command. The arrow indicates the direction the line must be drawn to ensure the orientation is correct (distal to the left) when rotated. D) Final rotated, straightened, cropped image. E) Cropped image with the large circle positioned tangent to the curve of the distal end of the gonad and the smaller circle placed around the center of the nucleus. Both circles are placed manually by user. F) Same image as in E, with features manually curated by user. G) Screenshot of spreadsheet output that results from "Measure all markups" command. H) Binary image that results from "Prepare for Sholl plugin" command. Note that for visual clarity, the contrast in this image has been inverted from the actual result in ImageJ. I) Same image as in H, with Sholl radii shown every 1µm. J) Screenshot of spreadsheet output from the Neuroanatomy plugin that runs the Sholl analysis (Ferreira et al. 2014). The screenshot has been trimmed to show the number of intersections that occur in the first 7 and final 3 radii. Worms imaged bear the qIs57[lag-2p::GFP] transgene (Seigfried et al., 2004; Davis et al., 2022). Scale bar is 10 µm in all images.

Picture from Browning H et al. (1996) Development "A sperm-supplied factor required for embryogenesis in C. elegans."

Figure 6. Immunofluorescence analyses of SPE-11 localization during spermatogenesis. In each confocal micrograph (A,C,D) spermatogenesis proceeds from left to right. SPE-11 is stained with affinity purified antiserum to SPE-11, DNA is stained with propidium iodide or 4, 6 diamidino-2-phenylindole (DAPI), andnuclear envelopes are stained with a monoclonal antibody (mAb414) to nuclear pore complex (NPC) proteins (Davis and Blobel, 1986). Scale bar, 10 µm. (A) Confocal images of a wild-type male gonad double labeled with antiserum to SPE-11 and propidium iodide. Each micrograph is the projection of a Z series containing seven sequential focal planes including approximately half the gonad thickness. SPE-11 first appears as dots in the nuclei of primary spermatocytes. The dots appear to coalesce before the first meiotic division. During the first and second meiotic divisions, SPE-11 is diffuse throughout the cells (bracket). SPE-11 is localized in a ring around the nuclei of sperm. (B) Conventional epifluorescence micrographs of sperm double labeled with antiserum to SPE-11 and DAPI. SPE-11 is perinuclear in sperm. Inset: Four spermatids attached to a single residual body. Some staining is detectable in the residual body. (C) Confocal images of primary spermatocytes double labeled with antiserum to SPE-11 and propidium iodide. The micrographs are projections of a Z series that includes the entire depth of the tissue. As the DNA becomes condensed in preparation for the first meiotic division, SPE-11 forms a fenestrated ring around the DNA. As discrete chromosomes become visible, SPE-11 relocalizes throughout the nucleus and is sometimes localized between the chromosomes. (This is also evident from analysis of individual focal planes; data not shown.) (D) Confocal images of primary spermatocytes double labeled with antiserum to SPE-11 and NPC antibodies. The micrographs are projections of a Z series that include the entire depth of the tissue. A merge of the NPC and SPE-11 projections demonstrates that the dynamic relocalization that SPE-11 undergoes in primary spermatocytes occurs within the nucleus. Spermatogenesis proceeds from left to right, with the exception of a few sperm (small rings stained with antiserum to SPE-11) in the upper left portion of the micrographs. There is no detectable staining of these sperm with the NPC antibody.