Picture from Yang Q et al. (2017) Dev Cell "The Invading Anchor Cell Induces Lateral Membrane Constriction during Vulval ...."

(M-P) Localization of the endogenous GFP::TOCA-1 reporter zh110 in wild-type (M) and unc-6(ev400) mutants (O and P) before (M and O) and after (N and P)lumen formation. (M') and (O') show yz projections of the lateral VulF membranes. The inset in (O) shows the displaced AC in the unc-6(ev400) mutant in a differentfocal plane.

Picture from Yang Q et al. (2017) Dev Cell "The Invading Anchor Cell Induces Lateral Membrane Constriction during Vulval ...."

Figure 6. The TOCA Proteins Are Required for Actomyosin-Driven Lateral VulF Constriction(A-F') Nomarski images (A-C, D-F) and CED-10::GFP expression (A'-C', D'-F') in mid-sagittal sections of wild-type (A-C') and toca-1(tm3334); toca-2 RNAi (D-F')larvae during vulval lumen formation. (A) and (D) show larvae stages before, (B) and (E) during, and (C) and (F) after lateral VulF constriction. The dashed red lines in (C) and (F) indicate the lengths of the lateral VulF membranes; the white arrowheads point to the gaps in the basal laminae formed by the invading AC, and the asterisks the positions of the AC. (G-H'') Localization of NMY-2::GFP in green and the Pcdh3>mCherry::PLCdPH reporter in magenta in wild-type (G-G'') and toca-1(tm3334);toca-2(ng11) doublemutant (H-H'') larvae at the Pn.pxx stage during vulval invagination. (G) and (H) are merged images of the mCherry::PLCdPH and NMY-2::GFP channels; (G'') and(H'') are yz projections of the lateral VulF membranes in the animals shown in (G') and (H').(I-J') Localization of the LifeAct::GFP F-actin reporter in wild-type (I) and toca-1(tm3334);toca-2(ng11) double-mutant (J) larvae at the Pn.pxx stage. (I') and (J')show yz projections of the lateral VulF membranes. In each panel, the yellow arrowhead points to the AC-VulF contact sites.(K) Intensity plots of NMY-2::GFP along the basal VulF membranes were generated from xy views as shown in the inset and described by Morf et al. (2013) (seealso STAR Methods). Error bars indicate the SD and the numbers in parentheses the numbers of animals analyzed for each genotype.(L) Intensity plots of Lifeact::GFP along the basal VulF membranes as described in (K). (M-P) Localization of the endogenous GFP::TOCA-1 reporter zh110 in wild-type (M) and unc-6(ev400) mutants (O and P) before (M and O) and after (N and P)lumen formation. (M') and (O') show yz projections of the lateral VulF membranes. The inset in (O) shows the displaced AC in the unc-6(ev400) mutant in a differentfocal plane.(Q-T) VPC-specific RNAi in the gfp::toca-1; rde-1(lf); rrf-3(lf) background. (Q) and (R) show control animals treated with empty vector and (S) and (T) are toca-1/-2RNAi-treated larvae. (Q and Q') and (S and S') show animals during constriction, while (R) and (T) are Nomarski images of the stage after constriction has beencompleted in the wild-type (after VulE division) used for the quantification shown in Figure S3J. Note in (S) the lack of GFP::TOCA-1 accumulation at the AC-VulFcontact sites and in (T) the abnormal lumen shape due to a lack of lateral VulF constriction indicated by the dashed red line.The asterisks mark the AC position and the yellow arrowheads point to the AC-VulF contact sites. Scale bars, 5 um.

Picture from Pineiro Lopez C et al. (2023) MicroPubl Biol "Segmentation of C. elegans germline nuclei."

Figure 1. Segmentation of germline nuclei in C elegans using Cell-ACDC and Cellpose:(A) Orthogonal views of a cropped z-stack of DAPI-stained C. elegans distal germline nuclei (left, imaged with AiryScan 880 microscope) were segmented with the cyto2 model of Cellpose (cyto2, purple, center) or a retrained model using 108 manually segmented "training" images (germlineNuclei, green, right). The fragmentation of individual nuclei is reduced with the custom model. White lines indicate x and y positions shown in the xz and yz orthogonal views. (B) Blurring by a factor of 2.5 (left: orthogonal views of blurred DAPI images) further improves the segmentation outcome for the cyto2 (center) and the custom germlineNuclei model (right). (C) The success of training is assessed by calculating the Jaccard index (Stringer et al., 2021; Pachitariu & Stringer, 2022) for 12 manually segmented "test" images. Jaccard indices for raw images are shown with open box plots, and with filled box plots for blurred images. Boxes show the interquartile range, medians are indicated by horizontal lines. Whiskers show smallest and largest value within 1.5 times the interquartile range below the 25th or above the 75th percentile, respectively. Blurring the data improves the Jaccard index for the cyto2 model (purple). By contrast, a re-trained custom model (m_0, blue) improves the segmentation of raw data but the Jaccard index is similar to the original cyto2 model for blurred data. Augmenting the training data improves the segmentation accuracy for both raw and blurred data sets (germlineNuclei, green). (D) The segmentation pipeline including all pre- and post-processing steps was implemented in Cell-ACDC (Padovani et al., 2022). Images shown are DAPI-stained C. elegans germline nuclei obtained with a SoRA spinning disk microscope.

Picture from Liegeois S et al. (2007) Genetics "Genes required for osmoregulation and apical secretion in Caenorhabditis ...."

Figure 7. CHE-14, VHA-5, and RDY-2 co-accumulate in the epidermis of vha-5 mutants affecting secretion but not in their sensory organs. (A) Fluorescence micrographs showing VHA-5::GFP (left) and RDY-2::GFP (right) fusion proteins in different adults (top), or VHA-5::CFP and RDY-2::YFP fusion proteins in the same L2 larva (bottom). Both proteins are expressed in the same tissues, i.e., the amphid sheath cell (top), the excretory cell and canal (middle), the epidermis (bottom, arrowheads). (Top) Lateral view. (Bottom) Ventral views. Bars, 10 um. (B) Confocal Z-projections through the epidermis (apical, top of each panel) showing the distributions of CHE-14::YFP and RDY-2::CFP in heterozygous (left) or homozygous (right) vha-5(mc38) adults carrying a vha-5(L786S)::mrfp mutant transgene (denoted Ex[L786S]). All three fluorescent proteins accumulate and colocalize (white arrrow) in homozygous vha-5(mc38) animals; the che-14 transgene was generally too weak to be detected in the excretory canal (yellow arrowhead). (C) Scheme of the C. elegans mouth (for the sake of simplicity, IL sensory organs are not represented), showing sensory organs where VHA-5 and RDY-2 are expressed. The XZ projection of confocal micrographs (bottom left) shows partial colocalization of VHA-5 and RDY-2 in the apical sheath cells of the amphids (larger dotted circles) and CEP (small dotted circles) and/or in the sheath cells of the OLQ sensory organs (arrowheads); two of the four CEP/OLQ organs seem to have no red signal because it was more difficult to detect them on the other side of the animal. (D) XY projections of confocal micrographs through the head of heterozygous (left) or homozygous (right) vha-5(mc38) adults carrying a vha-5(L786S)::mrfp mutant transgene, and YZ section through an amphid organ (corresponding to the dotted arrow in the XY projection). CHE-14 was uniquely expressed in the socket cell (to the left of the arrowhead; the sheath cell is to the right of the arrowhead). Five transgenic animals were examined in detail by confocal microscopy. Bars, 5 um.