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Image Contest 2021

Thank you to all our Image Contest Contributors!

Top three images:

Axolotl Soup by Dr. Marko Pende
Ultrastructure of a cell generating tension by Tejeshwar Rao, Ph.D.
The turtle shell by Frederic Bonnet, Ph.D.

Core Facility with the most users entries: MDIBL Light Microscopy Facility.

2nd Place
Ultrastructure of a cell generating tension – Tejeshwar Rao

3rd place
The turtle shell – Frederic Bonnet

Cos-7 cell generated tension – Tejeshwar Rao

Nourishing the mammalian egg – Sofia Granados Aparici

The Preimplantation Joker – Martin Kinisu

C elegans muscle fibers and ribosomal proteins – Hannah Somers

C elegans germline with p granules and ribosomal subunits tagged, DAPI stain – Hannah Somers

Axolotl soup – Marko Pende

Transgenic axolotl line showing components of the nervous system

Stereoskop: Zeiss Axio Zoom; Magnification: 11x

Light Microscopy Facility MDIBL

Ultrastructure of a cell generating tension – Tejeshwar Rao

The image is of a primary human umbilical vein endothelial cell (HUVEC) plated on a tension gauge tether (TGT) surface which allows visualization of the forces generated by the cell during attachment and spreading. The cells were further fixed and stained to visualize the underlying cytoskeletal architecture. The tension is highlighted in purple, the focal adhesions marked by paxillin staining in yellow and the cytoskeletal architecture in blue.

Nikon Ti2 was used to capture the image under the total internal reflection fluorescence mode of acquisition. The image was acquired with a 100X objective. The cells were plated on a artificially synthesized tension gauge tether (TGT) surface for 60 mins which allows visualization of single molecule integrin forces in the piconewton range upon fluorescent probe rupture. The underlying cytoskeletal proteins (paxillin and F-actin) were stained with antibodies.

The turtle shell – Frederic Bonnet

Pig kidney cells with Nuclei (white), Membranes (orange) and Vesicles (rainbow).

Image was taken with a widefield upright Zeiss AxioImager M2 equipped with Zeiss
Plan-Apochromat 63x/1.4 Oil objective. The DAPI, EGFP and Rhodamine Red were excited with a X-Cite 120 LED Boost fluorescent lamp and collected respectively with the DAPI – filter set 49, eGFP – filter set 38 and mCherry – filter set 63 HE. The 16 bit image was acquired with a Hamamatsu ORCA FLASH 4.0 LT controlled with Zen Pro 3.1 software

Light Microscopy Facility MDIBL

Black hole – Ravindra Zirmire

Spreading fibroblast stained with Phalloidin for actin

Olympus FV 3000, 60X objective

https://www.ncbs.res.in/research-facilities/ciff

Mucus Matters – Mindy Engevik

Here you can see a mouse colon producing robust amounts of mucus (purple) which keeps bacteria (yellow), undigested fibers (green) and other dietary components at a proper distance away from the intestinal epithelium (teal).

Zeiss Axio, 20x

Cos-7 cell generated tension – Tejeshwar Rao

An African green monkey kidney fibroblast Cos-7 cells plated on tension gauge tether surface with EGF stimulation. Cells are fixed and stained 60 mins post plating for focal adhesions (FAs) with the FA marker paxillin and the cytoskeletal marker F-actin. The underlying tension is shown in blue, the cytoskeletal organization in green and the focal adhesions in red.

Nikon Ti2 microscope was used in the total Internal reflection fluorescence (TIRF) configuration. The objective employed was a 60X oil immersion lens. The cells were plated on a 56 pN TGT surface which helps measure single molecule integrin forces in the piconewton range as the cell attaches and spreads. The cells were also stained post spreading with antibodies against paxillin and F-actin.

“Villi”-nous – Kristen Engevik

Immunofluorescence staining of adult mouse small intestine, showing membrane marker catenin in teal and nuclei visualized in purple

Imaged with a widefield epifluorescence Nikon TiE inverted microscope using a SPECTRAX LED light source (Lumencor) and a 10x Plan Fluor (NA 0.45) phase contrast objective

Infectious – Kristen Engevik

Immunofluorescence staining of mouse neonate small intestinal villi infected with rotavirus. Rotavirus infected cells are shown in magenta, cell membrane (catenin) are in purple and nuclei are visualized in white.

Imaged with a widefield epifluorescence Nikon TiE inverted microscope using a SPECTRAX LED light source (Lumencor) and a 40x objective

OrganoidFunk – Kristen Engevik

Immunofluorescence staining of 3D mouse gastric organoid expressing GFP-tagged actin (green), phalloidin is stained in magenta, nuclei visualized in blue and cleaved caspase 3 in red. The bright green cell in the right portion of the organoid was photodamaged by two-photon microscopy to cause single cell death, indicating increased recruitment of actin in neighboring cells to help exfoliate the damaged cell into the organoid lumen.

Imaged with Zeiss LSM 710 Live DUO confocal inverted microscope and a C-Achroplan NIR 40x objective. Photodamage was performed in a microscope incubation chamber on an Zeiss LSM 510 NLO confocal inverted microscope, where single cell damage is induced by repetitive scanning at high Ti-Sa laser power (~650 mW) for 500 iterations.

Flowering Colon – Mindy Engevik

Mouse Colon stained for actin (green), nuclei (pink) and adherent mucins (yellow)

Nikon Elcipse 20x

Innerspace Exploration – Aidyn Medina

Mouse Brain Microvascular endothelial cells that have been expose to spinal cord injury serum. Cells were live stain with Hoechst (Blue) for the nucleus and Mitotracker (Red) for mitochondria. After 3hrs of spinal cord injury serum exposure cells went through immunocytochemistry to label connexin 43 (Green).

Live Cell staining and microwave assisted immunocytochemistry.
Image acquisition using 3i/Olympus Inverted Epifluorescent Microscope with ZDC, a Lambda LS Xenon and a white LED light as dual excitation sources, and filter sets for 405, 488, 561 and 650 excitations. Images were captured using a Zyla sCMOS Digital camera (model 4.2CL10, Andor; Belfast, United Kingdom), SlideBook 6.0.17 Imaging Software. Magnification Olympus 150X 1.45 NA UApoN Oil Objective

Microscopy and Analytical Imaging Lab (MAI) at The University of Kansas

Nourishing the mammalian egg – Sofia Granados Aparici

The ovary is the female reproductive organ that contains the biggest cell in the human body: the egg. It is coated with a layer that prevents the attachment of more than one sperm during fertilization. On top of that, it is surrounded by one or more layers of cells in a structure called the ovarian follicle. This places the egg far from blood vessels and therefore with limited access to nutrients and oxygen. How then is the egg able to grow in a healthy manner? This is the role of the cells that surround it. They transfer the required nutrients by extending arm-like structures, called filopodia, that penetrate the coating layer and land on the egg surface, allowing the egg to increase in size and potentially become competent for fertilization. We investigate how filopodia are formed and elongated and how this process is crucial to maintain egg quality. This picture shows a high-resolution confocal microscopy image of an ovarian follicle. The egg is the big cell in the middle (in red) surrounded by a ring of smaller cells. The nucleus of each surrounding cell (containing the DNA) is shown in blue, including the nucleus of the egg, at the center. The filopodia formed by the surrounding cells (in green) can be appreciated in the space between these cells and the egg, corresponding to the coating layer. The red dots on these filopodia show a protein that allows their elongation towards the egg.

Zeiss LSM 780 laser scanning confocal microscope at 63x magnification

Molecular Imaging Platform of the Research Institute of the MUHC

Ocean Eye – Diana Djaksigulova

This is the cornea of a 3 month-old mouse which was heterozygous for the LacZ gene on the X-chromosome. The cornea was stained with x-gal to reveal the pattern of X-chromosome inactivation and the proliferation of cells into the center.

Olympus BX61 Bright Field Microscope was used, and imaged at 10x magnification and images were stitched together using the tile function

BC Children’s Hospital Imaging Core

The Best Mistake – Zbigniew Mikulski

Wholemount image of popliteal lymph node with macrophages (green) and lymphatic conduits (magenta). Mouse footpad was injected with a combination of anti-CD169 antibodies and a low molecular weight dye that was preferentially accumulating in conduits. We were hoping to label Zika virus with the dye, but the reaction did not work properly and we got this beautiful mistake instead. Image taken during collaboration with Willam Tang, from Sujan Shresta lab at LJI.

Leica SP5, 25x 0.95NA water immersion FLUOTAR objective. Imaging in confocal mode with 2 AU pinhole size on PMT-based detectors. Tiled Z-stacks were stitched and maximum intensity projection was done in Fiji.

La Jolla Institute for Immunology

Colonic mucus – Mindy Engevik

Human colon showing secreted MUC2 mucus (purple) and adherent mucus MUC1 (green), couterstained for nuclei (blue).

Zeiss Axio, 20x

The Cell Alter Ego – Aidyn Medina

Mouse Brain Microvascular endothelial cells that have been injured with spinal cord injury serum. Cells were live stain with Hoechst (Blue) for the nucleus and Mitotracker (Red) for mitochondria. After 3hrs of spinal cord injury serum exposure cells went through immunocytochemistry to label connexin 43 (Green).

Image acquisition using 3i/Olympus Inverted Epifluorescent Microscope with ZDC, a Lambda LS Xenon and a white LED light as dual excitation sources, and filter sets for 405, 488, 561 and 650 excitations. Images were captured using a Zyla sCMOS Digital camera (model 4.2CL10, Andor; Belfast, United Kingdom), SlideBook 6.0.17 Imaging Software. Magnification Olympus 150X 1.45 NA UApoN Oil Objective

Microscopy and Analytical Imaging Lab (MAI) at The University of Kansas

The Preimplantation Joker – Martin Kinisu

Single blastomere of a mouse two-cell stage embryo was injected with H2B-GFP mRNA and then allowed to culture to the blastocyst stage. Next the blastocyst was stained for trophectoderm marker Cdx2 (purple)

Nikon TE-2000

Stress granule – Nicolas Stifani

Stress granule yellow, DAPI cyan, actin filament magenta in eukaryote cells

Nikon E600 100x/1.3 Oil Color CMOS Nikon DS-Ri2 4908 x 3264 pixels,14-bit

Université de Montréal FacMed Microscopy

Sun – Ravindra Zirmire

It is spreading fibroblast stained with actin cytoskeleton in red and nucleus in blue.

Olympus FV 3000, 60X objective

CIFF BLiSC

C elegans muscle fibers and ribosomal proteins – Hannah Somers

A C elegans strain with a ribosomal subunit and muscle fiber fluorescently tagged

Nikon Ti-E Spinning Disk (60x oil objective was used), Imaris software was used to visualize and save 3D image.

MDIBL Light Microscopy Facility

C elegans germline with p granules and ribosomal subunits tagged, DAPI stain – Hannah Somers

This C elegans strain has ribosomal subunits and P granules fluorescently tagged, and includes a DAPI stain.

Zeiss LSM 980 was used to capture this image, using the 20x air objective. Imaris was used to create the 3D image.

MDIBL Light Microscopy Facility

Stars in the cell – Ravindra Zirmire

The image comprises fibroblast cell stained with Early endosome marker in red and actin cytoskeleton in green

IX 73 microscope, 40X magnification

CIFF BLiSC

The Walking Cells – Frederic Bonnet

Pig kidney cells with Nuclei (blue), Membranes (Magenta) and Vesicles (green)

The image was collected with a widefield upright Zeiss AxioImager M2 equipped with a Plan-Apochromat 63x/1.4 Oil objective. The DAPI, EGFP and Rhodamine Red fluorescence were excited with a X-Cite 120 LED Boost fluorescent lamp and collected respectively with the Zeiss filter DAPI – filter set 49, eGFP – filter set 38 and mCherry – filter set 63 HE. Image were collected in 16bit on an Hamamatsu ORCA FLASH 4.0 LT and with the Zen Pro 3.1 software.

MDIBL Light Microscopy Facility

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RRID: SCR_024409

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