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opencc-by-4.0Jan 2022View details →

Confocal Microscopy Visualizes Particle-Crack Interactions in Epoxy Composites with Optical Force Probe-Crosslinked Rubber Particles

Data (*.csv and *.lif) corresponding to Figures 2-7 of the manuscript and Figures S1-S2 of the Supporting Information.

opencc-by-4.0Oct 2018View details →

Confocal fluorescence microscopy images of the lacuno-canalicular network in bone femoral diaphysis of mice from the BionM1 project (space flight)

This data set provides complementary measurements to a separate THG data set of the same study: doi: 10.5281/zenodo.1475906 Data set for 1 sample of each of the 3 groups: Control, Space Flight and Synchro (ground control with space flight housing and feeding conditions). Contains confocal fluorescence microscopy images in tif format of 2D mosaic of selected samples and 3D stacks in selected anatomical regions of interest. See readme file for more information.

opencc-by-4.0Nov 2018View details →

Cisplatin enhances cell stiffness and decreases invasiveness rate in prostate cancer cells by actin accumulation: Confocal and atomic force microscopy

Summary Dataset of imaging data related to the publication  Raudenska, M., Kratochvilova, M., Vicar, T., Gumulec, J., Balvan, J., Polanska, H. Pribyl, J. & Masarik, M.:Cisplatin enhances cell stiffness and decreases invasiveness rate in prostate cancer cells by actin accumulation. Scientific Reports 2019, 9, 1660 This dataset includes image data of atomic force microcopy (Young modulus) and confocal microscopy(staining of F-actin and β-tubulin) of prostate cell lines PNT1A, 22Rv1, and PC-3.  Materials and Methods Cells, cell culture conditions Cells confluent up to 50–60% were washed with a FBS-free medium and treated with a fresh medium with FBS and required antineoplastic drug concentration (IC50 concentration for the particular cell line). The cells were treated with 93 µM (PC-3), 38 µM (PNT1A), and 24 µM (22Rv1) of cisplatin (Sigma-Aldrich, St. Louis, Missouri), respectively. IC50 concentrations used for treatment with docetaxel (Sigma-Aldrich, St. Louis, Missouri) were 200nM for PC-3, 70nM for PNT1A, and 150nM for 22Rv1.  Long-term zinc (II) treatment of cell cultures Cells were cultivated in the constant presence of zinc(II) ions. Concentrations of zinc(II) sulphate in the medium were increased gradually by small changes of 25 or 50 µM. The cells were cultivated at each concentration no less than one week before harvesting and their viability was checked before adding more zinc. This process was used to select zinc resistant cells naturally and to ensure better accumulation of zinc within the cells (accumulation of zinc is usually poor during the short-term treatment of prostate cancer cells). Total time of  the cultivation of cell lines in the zinc(II)-containing media exceeded one year. Resulting concentrations of zinc(II) in the media (IC50 for the particular cell line) were 50 µM for the PC-3 cell line, 150 µM for the PNT1A cell line, and 400 µM for the 22Rv1 cell line. The concentrations of zinc(II) in the media and FBS were taken into account.  Actin and tubulin staining β-tubulin was labeled with anti- β tubulin antibody [EPR1330] (ab108342) at a working dilution of 1/300. The secondary antibody used was Alexa Fluor® 555 donkey anti-rabbit (ab150074) at a dilution of 1/1000. Actin was labeled with Alexa Fluor™ 488 Phalloidin (A12379, Invitrogen); 1 unit per slide. For mounting Duolink® In Situ Mounting Medium with DAPI (DUO82040) was used. The cells were fixed in 3.7% paraformaldehyde and permeabilized using 0.1% Triton X-100.  Confocal microscopy The microscopy of samples was performed at the Institute of Biophysics, Czech Academy of Sciences, Brno, Czech Republic. Leica DM RXA microscope (equipped with DMSTC motorized stage, Piezzo z-movement, MicroMax CCD camera, CSU-10 confocal unit and 488, 562, and 714 nm laser diodes with AOTF) was used for acquiring detailed cell images (100× oil immersion Plan Fluotar lens, NA 1.3). Total 50 Z slices was captured with Z step size 0.3 μm. Atomic force microscopy We used the bioAFM microscope JPK NanoWizard 3 (JPK, Berlin, Germany) placed on the inverted optical microscope Olympus IX‑81 (Olympus, Tokyo, Japan) equipped with the fluorescence and confocal module, thus allowing a combined experiment (AFM‑optical combined images). The maximal scanning range of the AFM microscope in X‑Y‑Z range was 100‑100‑15 µm. The typical approach/retract settings were identical with a 15 μm extend/retract length, Setpoint value of 1 nN, a pixel rate of 2048 Hz and a speed of 30 µm/s. The system operated under closed-loop control. After reaching the selected contact force, the cantilever was retracted. The retraction length of 15 μm was sufficient to overcome any adhesion between the tip and the sample and to make sure that the cantilever had been completely retracted from the sample surface. Force‑distance (FD) curve was recorded at each point of the cantilever approach/retract movement. AFM measurements were obtained at 37°C (Petri dish heater, JPK) with force measurements recorded at a pulling speed of 30 µm/s (extension time 0.5 sec). The Young's modulus (E) was calculated by fitting the Hertzian‑Sneddon model on the FD curves measured as force maps (64x64 points) of the region containing either a single cell or multiple cells. JPK data evaluation software was used for the batch processing of measured data. The adjustment of the cantilever position above the sample was carried out under the microscope by controlling the position of the AFM‑head by motorized stage equipped with Petri dish heater (JPK) allowing precise positioning of the sample together with a constant elevated temperature of the sample for the whole period of the experiment. Soft uncoated AFM probes HYDRA-2R-100N (Applied NanoStructures, Mountain View, CA, USA), i.e. silicon nitride cantilevers with silicon tips are used for stiffness studies because they are maximally gentle to living cells (not causing mechanical stimulation). Moreover, as compared with coated cantilevers, these probes are very stable under elevated temperatures in liquids – thus allowing long-time measurements without nonspecific changes in the measured signal. Image analysis Fluorescence microscopy data were analyzed in ImageJ 1.52h and Python 3.7.1 as follows: cells were manually segmented using actin fluorescence channel, two regions were created for analysis: whole cell and cell periphery, lining a 4 μm thick region around cell border and including most of periphery actin cytoskeleton. In these two regions following parameters were measured for both actin and tubulin fluorescence: Integrated intensity, median intensity, and following regions were measured to describe cell morphology: Cell area, Maximum caliper (max feret diameter), roundness, and aspect ratio. Moreover, stress fibers were manually segmented in every cell and following parameters were measured: number of fibers per cell, feret angle of fiber, integrated intensity, fiber length, mean intensity. Next, a standard deviation of feret angles of individual fibers was calculated relatively to mean of feret angle using a circstd function from scipy package for Python. Identification of files Microscopy data Files are separated into individual zip files. The dataset of confocal microscopy is separated based on treatments: untreated control, docetaxel-treated cells, cisplatin-treated cells, zinc-treated cells. Filenames actin_tubulin_Zstack_cisplatin.zip, actin_tubulin_Zstack_untreated_control.zip, actin_tubulin_Zstack_zinc.zip, actin_tubulin_Zstack_docetaxel.zip. Files included in these ZIP archives are named as follows: "cellline_treatment_FOV". Files are 3-layer 16bit tiff files with layer sequence as follows: F-Actin (Phalloidin)/b-tubulin/Hoechst 33342. The dataset contains 242 FOVs of three cell line types/three treatments + one control, files are Z-stacks made of 50 slices. The dataset of atomic force microscopy (AFM) is included in one ZIP archive "AFM_YoungModulus_SetpointHeight.zip", which includes data on Young modulus and Setpoint Height of cell lines 22Rv1, PNT1A and PC-3 and treatments zinc, docetaxel, cisplatin (+control), i.e. identical like for confocal microscopy. The file naming is as follows: "AFM_cellline_treatment_FOV_Youngmodulus.tif"  for Young modulus and "AFM_cellline_treatment_FOV_setpointheight.tif" for setpoint height. The data are filtered 32-bit tiff images, where the pixel value correspond to cell stiffness (young modulus) in Pa or setpoint height in m. Confocal microscopy analysis files Following files are csv tables including image analysis of actin/tubulin staining captured by confocal microscope: Cytoskeleton_fluo_analysis_Cell_Cell_periphery_morphology.csv: table includes analyzed data for actin and tubulin staining in following cellular regions: cell, cell periphery. Standard ImageJ parameters regarding intensity and morphology included. Cytoskeleton_fluo_analysis_Fibers.csv: table includes results of manual segmentation and consequent analysis of actin stres fibers in the cells. Apart from standard ImageJ parameters, also number of stress fibers per cell and standard deviation of fiber angle relative to the cell mean angle (for details see methods) are included.

opencc-by-4.0May 2023View details →

Super-Resolved FRET Imaging by Confocal Fluorescence-Lifetime Single-Molecule Localization Microscopy

FRET-based methods are a special tool for detecting interactions between (bio)molecules and their immediate environment. The spatial distribution of molecular interactions and functional states can be seen using FLIM (Fluorescence Lifetime IMaging) and FRET imaging. The spatial information, accuracy, and dynamic range of the observed signals are, however, constrained by the fact that conventional FLIM and FRET imaging only provides average information over an ensemble of molecules within a diffraction-limited volume. On the other hand, conventional Single Molecule Localization Microscopy (SMLM) relies on highly sensitive multi-pixel detectors (e.g. sCMOS or EM-CCD) whose time resolution is not suitable for fluorescence lifetime measurements. Here, we demonstrate a method for obtaining super-resolved FRET imaging using confocal fluorescence-lifetime single-molecule localization microscopy. The proof of concept was carried out using a DNA origami sample for performing DNA-PAINT measurements in combination with fluorogenic probes for reducing background signal. With this method, We show that FRET events separated by sub-diffraction distances can be distinguished based on lifetime modifications.

opencc-by-4.0Apr 2016View details →

Fluorescent Confocal Laser Scanning Microscopy of White Blood Cells, Cancer Cell Line MCF7, and Mixtures of these Cells: A Model System for Circulating Tumor Cell Biomarker Evaluation V.1

This is a confocal laser scanning microscopy data set of white blood cells (leukocytes), the cancer cell line MCF7, and mixtures of these cells acquired on a Zeiss LSM 780 microscope in the University of Colorado Anschutz Medical Campus Advanced Light Microscopy Core. Cells are fluorescently labeled for DNA with DAPI (Sigma D9542), lipids with Bodipy 495/503 (Thermo Fisher D3922), the filament protein cytokeratin (CK) with pan-cytokertain-alexa555 antibodies (Cell Signaling Technologies 3478S) and the surface membrane antigen CD45 with CD45-alexa647 antibodies (Biolegend 304020). Bodipy was excited with a continuous wave (CW) 488 nm laser, alexa555 was excited with CW 561 nm laser, and alexa647 was excited with a CW 633 nm laser. The acquiring instrument does not have a CW 405 nm source so DAPI was excited by two photon process using a Coherent Cameleon ultrafast pulsed laser tuned to 765 nm. The objective used was a Zeiss Plan-Apochromat 20x, 0.8 NA, air. The data consists of 4 channel 8x8 mosaic z-stacks. The Zeiss software performed stitching of the mosaics. These stitched data images are included and marked with _Stitched at the end. Those interested in performing the stitching themselves can do this with the raw data files (without the _Stitched). The jpeg images are processed from the stitched LSM images. The LSM files contain additional meta data on the experiment including power levels and acquisition settings. The _Stiched .lsm files will load in ImageJ (tested with V.1.49) as 4 channel 3 stack images. This data is a model system for evaluating the DNA/Lipids/CK/CD45 biomarker panel to identify circulating tumor cells (CTCs). The D- population of the model is the WBCs and the D+ population is the MCF7 cancer cell line. The amount of separation the biomarker panel plus analysis algorithm can produce between these populations (D+/D-) is an estimate the sensitivity and specificity of the biomarker panel plus algorithm to CTCs. Experiments generating the data were performed over the course of 15 days. Peripheral blood samples were collected from the Gynecological Tissue and Fluid Bank (COMIRB 07-0935 / COMIRB 05-1081) from consenting patients undergoing surgery at the University of Colorado Hospital. Blood samples were used the same day they were collected. Blood samples were collected from 3 patients with benign conditions, labeled WBBN#, and 3 patients with ovarian cancer, labeled WBCA#. We do not expect there to be any difference in the isolated white blood cells samples prepared from the cancer and benign patients. Samples were stored at room temperature until white blood cells were isolated. Mixed samples were prepared by passaging a MCF7 flask and mixing it with isolated white blood cells before fixation. A schedule showing the time duration between collection, processing and imaging is included as “experimental schedule.gif”. The MCF7 cancer cell line was a kind gift from Dr. Heide Ford. Genomic DNA was isolated from the MCF7 cell line after the experiment and sent for cell line authentication. The gDNA was a match to MCF7. The authentication report and data are included in this submission. CD45 antibodies were exhausted on day 7. New antibody was purchased and received on day 8. The day 7 images only has labels for DAPI and Bodipy. The samples prepared with the old antibodies on days 4 and 7 were relabeled and imaged with the new antibodies on days 14 and 15. This labeling was also done to confirm the pan-CK antibodies remained good since they are dim in the MCF7 cells imaged on days 12 and 13. The pan-CK on days 14 and 15 looks the same as it did on days 5 and 7 confirming the antibodies are good. Four of the filters containing cells were not sufficiently flat to be acquired with a 3 slice z-stack so a 5 slice z-stack was used. These files have been zipped to compress them under the 2 GB limit permitted by zenodo.org Further information on how these samples were prepared, processed, and analyzed can be found in our associated 2016 SPIE Photonics West BIOS conference proceeding titled, “Quantitative image cytometry measurements of lipids, DNA, CD45 and cytokeratin for circulating tumor cell identification in a model system”, http://dx.doi.org/10.1117/12.2222317. This work was supported by funding provided to the University of Colorado Cancer Center by the American Cancer Society and awarded as Institutional Research Grant Number 57-001-53, by funding provided by the Defense Advanced Research Projects Agency under grant number N66001-10-4035, and by funding provided by NIH/NCATS Colorado CTSI Grant Number TL1 TR001081. The University of Colorado Anschutz Medical Campus Advanced Light Microscopy Core is also supported in part by NIH/NCATS Colorado CTSI Grant Number UL1 TR001082. The funders had no role in the study design, data collection, analysis, or decision to publish.

opencc-by-4.0Sep 2023View details →

Supplementary material - Optical Diffraction Tomography and Raman Confocal Microscopy for the Investigation of Vacuoles Associated with Cancer Senescent Engulfing Cells

Supplementary material containing the data used in the manuscript "Optical Diffraction Tomography and Raman Confocal Microscopy for the Investigation of Vacuoles Associated with Cancer Senescent Engulfing Cells"

opencc-by-4.0Nov 2024View details →

Time-lapse 3D confocal microscopy videos of mitochondrial dynamics in human alveolar epithelial cells (A549-DsRed) infected with Mycobacterium marinum (Mmar) strains

<div>The dataset consists of time-lapse, 3D confocal images of mitochondrial dynamics in human alveolar epithelial cells (A549-DsRed) infected with Mycobacterium marinum (Mmar) strains. Images were captured at 60X magnification in an environmental chamber at 35°C for live-cell imaging. Host cell mitochondria were labeled with red fluorescent protein (RFP) and infected with both wildtype (wt) and ESAT-6 operon knockout mutant labeled with green fluorescent protein (GFP) at MOI of 100 for 24 hours at 35°C. Infected cells were identified and analyzed to explore the effect of pathogenic mycobacteria on mitochondrial morphology over time.</div> <div> </div> <div>More details available in this preprint: <a href="https://doi.org/10.48550/arXiv.2411.06035">https://doi.org/10.48550/arXiv.2411.06035</a></div>

opencc-by-4.0Mar 2024View details →

Correlative microscopy of mice cerebellar Purkinje cells from 20x confocal tissue imaging to super-resolution 93x 3D STED of dendritic spines

This Dataset concerns the paper entitled "From tissues to segmentation: a modular framework for multi-scale neuron isolation" by Cauzzo et al. Nature Comm (2024). S.Cauzzo$, E. Bruno, D. Boulet, P. Nazac, M. Basile, A. L. Callara, F. Tozzi, A. Ahluwalia, C. Magliaro, L. Danglot$*, N. Vanello$*    *shared senior authorship: Lydia.danglot@inserm.fr ; nicola.vanello@unipi.it $ corresponding authors : cauzzo.simone@gmail.com  ; Lydia.danglot@inserm.fr ; nicola.vanello@unipi.it

opencc-by-4.0Oct 2023View details →

Confocal microscopy images of Escherichia coli cells treated with various antibiotics (LB, exponential phase)

This dataset and CARE model is part of the publication "Transertion and cell geometry organize the Escherichia coli nucleoid during rapid growth".It contains all CLSM images that were used for the publication, as well as the single-cell regions of interest for analyses.Cells were grown to exponential phase in LB Lennox and antibiotics were added for 0-60 min. Cultures were then chemically fixed, immobilized and stained for DNA (DAPI) and membrane (Nile Red). The strain (NO34) expresses a MreBsw-sfGFP fusion protein from the native chromosomal locus. It was a kind gift from Zemer Gitai (<a href="http://doi:10.1016/j.bpj.2016.07.017">Ouzounov et al., 2016</a>).More information can be found in the publication.

opencc-by-4.0Feb 2024View details →

Metadata for Confocal Laser Scanning Microscopy Images of Monoculture and Mixed-Species Biofilms Formed by Bacterial Isolates of Dairy Origin

In a project conducted by ILVO (Belgium), a wide variety of bacterial species were recovered from the surface of a dairy pasteurizer after cleaning and disinfection (C&D). The biofilm-forming ability of these bacteria was determined in both single-species and various mixed-culture combinations. Some work related to this study has been published in Frontiers: "Synergistic interactions in multispecies biofilm combinations of bacterial isolates recovered from diverse food processing industries". Bacterial species were mixed in different combinations to assess the community biofilm mass and growth dynamics of individual species. ILVO and the University of Copenhagen conducted experiments aimed at revealing the structural characteristics and spatial organization of bacterial species within different mixed-species biofilms. In our research, we employed oligonucleotide FISH probes, each conjugated with a unique fluorescent dye: Cy5 for Stenotrophomonas rhizophila (B68), Cy3 for Bacillus licheniformis (B65), and FAM for Microbacterium lacticum (B30). C1 combination refers to a combination containing B68 and B30.  Images of the biofilms formed on the coupons were captured using a confocal laser scanning microscope (LSM 800, Zeiss) with a Plan-Apochromat 63x/1.4 oil-immersion objective. Z-stacks were recorded to obtain three-dimensional (3D) images. Standard images were made with an image size of 1024 × 1024 pixels, corresponding to physical dimensions of 101.4 × 101.4 μm for each image. For each image, two separate channels were applied to detect any dual-species combination using a flexible detector (GaAsP-PMT) in the LSM 800 system. Representative 3D views of images were generated using the 3D model function in the ZEN system 3.7. Biofilms were grown in BHI for 24 h on plastic coupons. The samples were imaged at different time points: 6h, 12h, 18h and 24h. Each samples had three replicates and for each replicate imaging was performed from 3-6 different positions.  Details of the oligonucleotide probes are given below: <table> <tbody> <tr> <td> Name of the species </td> <td> Sequences </td> <td> Max. excitation </td> <td> Max. emission </td> <td> Fluorophores </td> </tr> <tr> <td> S. rhizophila B68  </td> <td> GGGCCTTTACCCCGCCA </td> <td> 649 nm </td> <td> 670 nm </td> <td> Cy5 </td> </tr> <tr> <td> B. licheniformis B65 </td> <td> ACCGCCTGCGCGCGCTT </td> <td> 550 nm </td> <td> 570 nm </td> <td> Cy3 </td> </tr> <tr> <td> M. lacticum B30 </td> <td> CCCCACCCTTTCGCTCC </td> <td> 495 nm </td> <td> 520 nm </td> <td> FAM </td> </tr> </tbody> </table>

opencc-by-4.0Apr 2024View details →

Correlative microscopy of rat cultured hippocampal pyramidal cell from 40x confocal imaging to super-resolution 93x 3D STED of dendritic spines

This dataset contain multi-scale image of rat hippocampal pyramidal cell related to our paper "From tissues to segmentation: a modular framework for multi-scale neuron isolation" by Cauzzo et al. Nature Comm (2024).

opencc-by-4.0Feb 2019View details →

Dataset of confocal microscopy stacks from plant samples - ImageJ SurfCut: a user-friendly, high-throughput pipeline for extracting cell contours from 3D confocal stacks

This data set contains confocal stacks from Arabidopsis thaliana 35S::GFP-MBD light grown hypocotyl as well as propidium iodide stained cotyledon pavement cells and shoot apical meristem. This is the test dataset for the Fiji macro SurfCut (https://github.com/sverger/SurfCut; 10.5281/zenodo.2635737)   Material and methods: Plant material and growth conditions Arabidopsis thaliana wild type Col-0 and the microtubule reporter line GFP-MBD (WS-4, (Marc et al. 1998) were used. Seeds were cold treated for 48 hr to synchronize germination. Plants were then grown in a phytotron at 20°C, in a 16 hr light/8 hr dark cycle on solid Murashige and Skoog medium (MS medium, Duchefa, Haarlem, the Netherlands) with 0.8% agar, 1% sucrose, and no vitamin.   Confocal microscopy Cell contour staining in the case of PC_PI_Col0_(1-8).tif and SAM_PI_Col-0.tif was performed by staining the cell wall with Propidium Iodide (PI). Plants were immersed in 0.2 mg/ml propidium iodide (PI, Sigma-Aldrich) for 10 min and washed with water prior to imaging. For imaging, samples were either placed on a solid agar medium and immersed in water, or placed between glass slide and coverslip separated by 400 μm spacers to prevent tissue crushing. Images were acquired using a Leica TCS SP8 confocal microscope, equipped with a water immersion objective (HCX IRAPO L 25x/0.95 W). PI excitation was performed using a 552 nm solid-state laser and fluorescence was detected at 600–650 nm. GFP excitation was performed using a 488 nm solid-state laser and fluorescence was detected at 495–535 nm. Stacks of 1024x1024 pixels (pixel size of 0.363 x 0.363 micron) optical section were generated with a Z interval of 0.5 μm.   File list: Light grown hypocotyl, GFP-MBD reporter line: - Hypocotyl_GFP-MBD.tif Cotyledon’s pavement cells, PI staining: - PC_PI_Col0_1.tif - PC_PI_Col0_2.tif - PC_PI_Col0_3.tif - PC_PI_Col0_4.tif - PC_PI_Col0_5.tif - PC_PI_Col0_6.tif - PC_PI_Col0_7.tif - PC_PI_Col0_8.tif Shoot apical meristem, PI staining: - SAM_PI_Col-0.tif   Reference: Marc, Jan, Cheryl L. Granger, Jennifer Brincat, Deborah D. Fisher, Teh-hui Kao, Andrew G. McCubbin, and Richard J. Cyr. 1998. “A GFP–MAP4 Reporter Gene for Visualizing Cortical Microtubule Rearrangements in Living Epidermal Cells.” The Plant Cell 10 (11): 1927–39. https://doi.org/10.1105/tpc.10.11.1927.

FIG. 4 in New insights on the morphology of a digenean parasite Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927)) using confocal laser scanning microscopy

FIG. 4. — Confocal tomographies of the reproductive system of Brachylaima mazzantii (Travassos, 1927): A, Mehlis' gland (mg), vitelline reservoir (vr), ovary (ov) and testes (t); B, ovary (ov), seminal reservoir (sr), testes (t) and intestinal caeca (ic); C, ovary (ov) and ootype (oo); D, vitelline glands (vg) forming lobed acini; E, vitelline duct (vd) showing vitelline cells inside forming a single row; F, uterus full of eggs; G, egg, revealing the embryo (emb), eggshell (sh), operculum (op) and discontinuity area (da) in the eggshell. Scale bars: A, B, D, F, 50 μm; C, E, 10 μm; G, 5 μm.

FIG. 8 in New insights on the morphology of a digenean parasite Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927)) using confocal laser scanning microscopy

FIG. 8. — Reproductive system of Brachylaima mazzantii (Travassos, 1927) as first described by Travassos in 1927 (adapted from Lent & Freitas 1937). Scale bar: 1 mm.

FIG. 1 in New insights on the morphology of a digenean parasite Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927)) using confocal laser scanning microscopy

FIG. 1. — Confocal tomographies of the reproductive system of Brachylaima mazzantii (Travassos, 1927): A, gland cells (gc) surrounding the genital pore (gp); B, commissure (co) and gland cells (gc); C, region of the genital opening showing differentiated musculature (dfm); D, female genital opening (fgo), male genital opening (mgo) and unarmed cirrus (ci); E, cirrus pouch (cip), metraterm (m), testes (t), bursa (b) and seminal vesicle (sv); F, vitelline duct (vd), vitelline reservoir (vr), uterus (u), ovary (ov) and testes (t). Scale bars: 50 μm.

FIG. 6 in New insights on the morphology of a digenean parasite Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927)) using confocal laser scanning microscopy

FIG. 6. — Confocal tomographies of tegument, musculature of the body and acetabulum of Brachylaima mazzantii (Travassos, 1927): A, tegument covered by many scales; B, scales; C, circular musculature (cm); D, longitudinal (lm) and diagonal musculature (dm); E, two differentiated muscle bundles (dfm); F, surface of acetabulum featuring many papillae (p); G, papillae (p), radial musculature (rm) and differentiated musculature (dfm) supporting the acetabulum; H, papillae. Scale bars: A-G, 50 μm; H, 10 μm.

FIG. 3 in New insights on the morphology of a digenean parasite Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927)) using confocal laser scanning microscopy

FIG. 3. — Schematic drawings of the reproductive system of Brachylaima mazzantii (Travassos, 1927): A, reconstruction of the reproductive system from the confocal tomographies, showing seminal vesicle (sv), metraterm (m), cirrus (c), cirrus pouch (cp), genital atrium (ga), testes (t), ovary (o), ootype (oo), vitelline duct (vd), vitelline glands (vg) vitelline reservoir (vr), and Mehlis' gland (mg); B, cirrus pouch as first described by Travassos in 1927 (adapted from Lent & Freitas 1937). Scale bars: A, 0.06 mm; B, 0.25 cm.

FIG. 5 in New insights on the morphology of a digenean parasite Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927)) using confocal laser scanning microscopy

FIG. 5. — Confocal tomographies showing the anterior region of Brachylaima mazzantii (Travassos, 1927): A, image showing the meridional musculature (mm); B, equatorial musculature (eqm); C, radial musculature (rm), papillae (p) on the surface of the oral sucker and differentiated musculature (dfm) making the transition between mouth and pharynx (pre-pharynx); D, pharynx (ph), radial musculature (rm), esophagus (e), and intestinal caeca (ic); E, detail of the intestinal caeca, revealing numerous microvilli (mi), and the epithelium (ep); F, esophageal glands (eg) circling the esophagus. Scale bars: A-D, 50 μm; E, 10 μm.

FIG. 2 in New insights on the morphology of a digenean parasite Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927)) using confocal laser scanning microscopy

FIG. 2. — Schematic drawings of the genital atrium of Brachylaima mazzantii (Travassos, 1927): A-D, sequential tomographic images starting from the body surface. Scale bar: 5 µm.

FIG. 7 in New insights on the morphology of a digenean parasite Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927)) using confocal laser scanning microscopy

FIG. 7. — Confocal tomographies of the excretory and nervous system of Brachylaima mazzantii (Travassos, 1927): A, excretory ducts (exd), testes (t); B, excretory bladder (exb), excretory pore (exp), and testes (t); C, longitudinal nervous cord (nc); D, commissures (co) originated from the nervous cord (nc). Scale bars: A-C, 50 μm; D, 10 μm.