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Image Contest 2022

Thank you to all our 2022 Image Contest Contributors!

Top three images:

An Arabidopsis embryo epidermis being patterned – Margot Smit
dinosaur bone – Michael Shribak
Brainstorm – Annika Lea Schulz

Core Facility with the most users entering: MDIBL Light Microscopy Facility

The People’s Choice Award is: Fruit fly ovary montage – Wen Lu and Vladimir I. Gelfand

An Arabidopsis embryo epidermis being patterned – Margot Smit

Fruit fly ovary montage – Wen Lu and Vladimir I. Gelfand

Reactive Brown Adipocytes – Maria Cecilia Marcondes

Microtubule and actin crosslinking is essential for oocyte growth – Wen Lu and Vladimir I. Gelfand

Every fruit fly starts with an oocyte – Wen Lu and Vladimir I. Gelfand

Red, White, and Blue Feather – Jenny C. Schafer

Developing Waxing Gibbos Moon – Arlen Ramírez Corona

Little monsters from the deep – Marko Pende

Sunrise of Embryonic Development – Lorenzo Gallicchio

Do not cross your own domain! – Lorenzo Gallicchio

When? Where? and How much? – Lorenzo Gallicchio

Lipid string coiled around mouse sperm flagellum – Alejandro Linares

Motoneuron in the trunk of a zebrafish – Romain Menard

Motoneurons synapse in muscle tissues of a zebrafish trunk – Romain Menard

Cobblestones of the skeletal muscle – Elham Javed

UFO, unidentified flying object – Aissette Baanannou

A broken heart – Josué David Hernández Varela

An Arabidopsis embryo epidermis being patterned – Margot Smit

Stanford University/HHMI

A plant (Arabidopsis) embryo that is forming a pattern on its leaf epidermis. Here we see and embryo with two leaves, the root is cut off on the left. The smaller, triangular cells are precursors for stomata, pores on the leaf surface through which the plant exchanges gasses with the environment. In cyan we’re seeing a signaling factor (EPF2), in purple/orange we see the receptor (TMM) to which the signal can bind. Together these two make sure the small cells are not formed next to each other and form a controlled, efficient pattern. This plant embryo is about 11 days old and will grow for another 4-6 days before desiccating and remaining dormant until germination.

Sample: Arabidopsis, embryo epidermis
Label: Epidermal plasma membrane was labeled with mCherry (grey), TMM protein with YFP (fire), EPF2 protein with CFP (cyan)
Instrument: Confocal Microscopy, Leica Stellaris, 25x objective
Scale: This image represents 0.6 mm x 0.6 mm
Image or sample processing steps:
This is a maximum projection of intensities from a z-stack that was 55 um think with a 1um step size. Brightness and contrast were adjusted for each channel and channels were merged in FIJI

dinosaur bone – Michael Shribak

A dinosaur bone in transmitted light under polychromatic polarization microscope (https://www.nature.com/articles/srep17340)). We used a 4x objective lens and illuminating white polarized light with a spectral fan of polarization ellipses. The picture shows real colors, as it is seen by naked eye.

Sample: tissue
Label: label-free
Instrument: polychromatic polarizing technique, Olympus IX81, 4x/0.13NA objective lens
Scale: 2.1 mm x 1.6 mm

Brainstorm – Annika Lea Schulz

Life Sciences Institute Imaging – Advanced Bioimaging Core at University of British Columbia

Organoid-Based COVID-19 Research: Identifying Potential Therapeutic Agents Against SARS-CoV-2 Using Lab-Grown Mini Brains. Organoid models are emerging as valuable tools to investigate viral biology and disease progression of COVID-19 The neuro-serine protease inhibitor (neuroserpin) Spn4A, is an antiviral biologic discovered in the Jean Lab (Department of Microbiology & Immunology, Life Sciences Institute, University of British Columbia) currently being explored as a SARS-CoV-2 entry inhibitor. Spn4A is a highly potent picomolar serpin of the human furin-like proteases. Here, the biological and antiviral activities of adenoviral-vector mediated Spn4A expression (shown in cyan) are investigated in a human induced pluripotent stem cell-derived dorsal forebrain organoid. Shown in red, is the furin proprotein convertase, which is hijacked by a multitude of viruses and, therefore, presents a promising target for broad-spectrum antiviral therapeutics. In addition, the transmembrane serine protease 2 (TMPRSS2, in green), another key host cell entry factor of SARS-CoV-2, is presented to indicate potentially susceptible cells (cell nuclei shown in blue) in the cerebral organoid.

Sample: Human induced pluripotent stem cell-derived dorsal forebrain organoid with adenoviral-vector mediated expression of neuro-serine protease inhibitor (neuroserpin) Spn4A. Clearing was performed after immunofluorescence staining using fructose-glycerol clearing reagent.
Label: The cell nuclei (DNA) were stained with the fluorescent dye Hoechst – Pseudocolour: blue. Furin was labelled with primary antibody: anti-furin polyclonal antibody (PA1-062) – Thermo Scientific & secondary antibody: gt-anti-rb Alexa647 – Pseudocolour: red. Transmembrane Serine Protease 2 (TMPRSS2) was labelled with primary antibody: anti-TMPRSS2 antibody (clone P5H9-A3, Cat. No. MABF2158) – Sigma Aldrich & secondary antibody: gt-anti-ms Alexa488 – Pseudocolour: green. The neuro-serine protease inhibitor was expressed using an adenoviral vector labelled with red fluorescent protein – Pseudocolour: cyan.
Instrument: Leica SP5 Inverted Confocal Microscope; 63X silicon oil immersion; tile scan protocol

Fruit fly ovary montage – Wen Lu and Vladimir I. Gelfand

Northwestern University Feinberg School of Medicine

Confocal images of wild-type fruit-fly ovaries (in the center) and dynein knockdown fruit-fly ovaries (on the periphery). Oocytes are the cells with very little DAPI staining, while the sister cells are the ones with the big nucleus of strong DAPI staining. The image shows that lack of the main microtubule minus-end directed motor, dynein, causes oocyte growth defects.

Sample: Drosophila melanogaster, ovaries, fixed with 4% formaldehyde
Label: DNA labeled with DAPI staining, grey; actin cytoskeleton labeled with rhodamine-phalloidin staining, pseudo-colored in cyan in the center, and blue, magenta, yellow and green on the periphery.
Instrument: Laser scanning confocal microscopy, Nikon A1R, 20X air lens, N.A.=0.75
Scale: 5 mm x 4 mm
Image or sample processing steps:
Maximum projection of Z-stack with 1um sectioning; images were processed in FIJI and assembled in Illustrator.

Not in Kansas anymore – Hannah Somers

MDIBL Light Microscopy Facility at MDI Biological Laboratory

The C. elegans displayed allow us to better understand how protein translation changes with age, and in specific tissues in whole animals.

Sample: C. elegans whole worm, fixed using PFA
Label: O-propargyl-puromycin Alexa647 labeling active translation, Dendra2 fluorescently tagged mechano-sensory neurons
Instrument: Zeiss LSM 980, confocal light imaging, 20x objective.
Scale: Scale is 2.5 mm x 1 mm
Image or sample processing steps:
Each of the worms were taken using a Z-stack that was about 100 um thick using 5 um sections, and 3D rendered using IMARIS software.

DNA painting – Marcel Mettlen

Quantitative Light Microscopy Core at UTSouthwestern Medical Center

The precision of today’s microscopes allows to direct and blink laser light with such precision that you can “paint” the Mona Lisa within ms inside the nucleus of a living cell.

Sample: U2OS
Label: U2OS were transfected with DNA coding for H2B-PSmOrange
Instrument: Nikon CSU-W1 with FRAP module; Objective: 100x; 1.45NA
Scale: in image

Tidepool Treasure – Hannah Somers

MDIBL Light Microscopy Facility at MDI Biological Laboratory

A beautiful sea star that can be found in lots of nooks and crannies around coastal New England.

Sample: Asterias amurensis (Northern Sea Star)
Label: Nuclear staining with DAPI in Cyan and auto-fluorescence in green
Instrument: Zeiss Axiozoom v16, EPI fluorescence imaging, 1x Objective.
Scale: 5 mm x 3 mm
Image or sample processing steps:
A z-stack and tiling were using to capture the image, the z-stack was about 1000 um thick, imaged every 50 um.

Reactive Brown Adipocytes – Maria Cecilia Marcondes

La Jolla Institute for Immunology at San Diego Biomedical Research Institute

Becoming Reactive

Sample: Mouse Brown Adipose Tissue
Label: Actin labelled with FITC-Phalloidin, mitochondria labelled with YFP-TOMM20, Superoxide labelled with 2-hydroxyethidium (DHE) and nucleus labelled with DAPI
Instrument: Zeiss Axiovert 200M, 40X magnification
Image or sample processing steps:
This is a primary culture of brown adipose tissue following exposure to Methamphetamine (1uM), a stimulant addictive drug, following a 15-minute pretreatment with DHE for superoxide detection. This image was performed 5 minutes after exposure to the drug. Fluorescent labelling was performed in PFA-fixed and Triton-X permeabilized cells, for co-localization of superoxide and mitochondria. Z-stack flattened in Image J/ FIJI

Microtubule and actin crosslinking is essential for oocyte growth – Wen Lu and Vladimir I. Gelfand

Northwestern University Feinberg School of Medicine

The image shows that wild-type Drosophila oocytes grow over time (yellow), while knockdown of an actin-microtubule cross-linker, Short stop (Shot), leads to a complete arrest of oocyte growth (cyan and magenta). In the background, actin filaments and microtubules are labeled in white in the shot mutant

Sample: Drosophila melanogaster, ovary tissue, fixed with 4% formaldehyde
Label: Actin filaments labeled with Rhodamine-Phalloidin staining, and pseudo-colored in yellow, cyan and magenta; in the background, actin filaments labeled with Rhodamine-Phalloidin and microtubules labeled with GFP-Patronin, both shown in white.
Instrument: Laser scanning confocal and spinning disk confocal; Nikon A1R and Nikon W1; 20X 0.75 N.A. air lens, ; 40 × 1.25 N.A. silicone oil lens
Image or sample processing steps:
The images were processed in FIJI and assembled in Adobe Illustrator.

Every fruit fly starts with an oocyte – Wen Lu and Vladimir I. Gelfand

Northwestern University Feinberg School of Medicine

Fruit-fly oocytes (labeled in magenta) grow over time in an assembly line within the ovary, to eventually give rise to mature eggs for starting new generations of fruit flies.

Sample: Drosophila melanogaster, ovary tissue, fixed with 4% formaldehyde
Label: DNA labeled with DAPI staining in cyan and oocytes labeled with anti-Orb antibody staining in magenta
Instrument: Laser scanning confocal microscope; Nikon A1R; 20X 0.75 N.A. air lens
Image or sample processing steps:
Maximum projections of Z-stacks of a total of 30-50 um thickness with 1 um sectioning; Images of various samples were processed in FIJI and assembled in Adobe Illustrator.

Red, White, and Blue Feather – Jenny C. Schafer

Vanderbilt Cell Imaging Shared Resource (CISR) at Vanderbilt University

Lost bird feather found in Vanderbilt parking garage and imaged for fun with autofluorescence to show 3D and XY tiling.

Sample: Feather found in the parking garage and imaged through a #1.5 coverslip. No sample preparation.
Label: Autofluorescence
Instrument: Zeiss LSM880 Confocal, 20x/0.8
Scale: 1.57 mm x 0.425 mm
Image or sample processing steps:
Maximum Intensity Projection of 29 slices over 64.375 um (2.3 um sectioning) and 4 horizontal tiles. Intensity adjustments for artistic effect and coloring were made in FIJI.

Pollen Grain – Jenny C. Schafer

Vanderbilt Cell Imaging Shared Resource (CISR) at Vanderbilt University

Pollen grain imaged by super-resolution and rendered in 3D to visualize shape and surface

Sample: Pollen grain slide from Carolina Biologicals. Fixed and mounted on slide with coverslip.
Label: Autofluorescence with 561 nm excitation
Instrument: Zeiss LSM880 Confocal with Airyscan, 20x/0.8
Scale: 85 um x 85 um
Image or sample processing steps:
Maximum Intensity Projection of 55 Z slices over 23.47 um (0.427 um sectioning). Airyscan processing and MIP created and pseudocolored in Zeiss ZEN software.

A perfect circle – Jenny C. Schafer

Vanderbilt Cell Imaging Shared Resource at Vanderbilt University

The actin cytoskeleton forms a perfect and beautiful circle around this cell.

Sample: Fluo cells from commercial slide, fixed and mounted
Label: Only actin channel shown. Fluo cells – MitoTracker568 for mitochondria, Alexa488-Phalloidin for actin, DAPI for nuclei
Instrument: Zeiss LSM710 Confocal, 63x/1.4
Scale: 107 um x 107 um
Image or sample processing steps:
Airyscan processing in Zeiss ZEN. Maximum Intensity Projection of 13 slices over 4.5 um (0.37 um sectioning). MIP rendered and pseudocolored in Zeiss ZEN.

Developing Waxing Gibbos Moon – Arlen Ramírez Corona

Laboratorio Nacional de Microscopía Avanzada, UNAM at Universidad Nacional Autónoma de Mexico

A lateral view of the blastoderm cells migrating over the surface of the yolk cell during epiboly in a zebrafish embryo. The nuclei of the cells are visible in magenta and the actin cytoskeleton at the cell borders is visible in cyan.

Sample: Zebrafish embryo fixed at 60% epiboly. Image shows the blastoderm migrating over the surface of the yolk cell
Label: Actin cytoskeleton labeled with Alexa488-phalloidin (cyan) and nucleus labeled with DAPY (magenta).
Instrument: Confocal microscopy. Confocal Olympus FV100, type IX81 Inverted. Objective UPLSAPO 10X/0.4
Scale: This image represents 848 x 848 um
Image or sample processing steps:
This is a maximum projection of intensities from a z-stack 29 slices thick. The projection was done using FIJI.

Cells on the run – Arlen Ramírez Corona

Laboratorio Nacional de Microscopía Avanzada, UNAM at Universidad Nacional Autónoma de Mexico

Migrating front of the blastoderm cells on the sourface of the yolk cell. Cell borders are labeled in cyan (actin), nuclei labeled in magente (DAPI) and E-cadherin is labeled in yellow.

Sample: Zebrafish embryo fixated at 60% epiboly.
Label: Actin cytoskeleton labeled with Alexa488-phalloidin, Nuclei labeled with DAPI and E-cadherin labeled with secondary antibody coupled with Alexa647.
Instrument: Olympus FV1000 type IX81 Inverted Confocal Microscope. Confocal Microscopy. UPLSAPO 60XS/1.3
Scale: This image represents 140.8 x 140.8 um
Image or sample processing steps:
This is a maximum projection of intensities from a z-stack that was 30 slices thick. The image was created using FIJI.

Axolotl space ship – Marko Pende

MDIBL Light Microscopy Facility at MDIBL

Axolotl Mothership

Sample: Axolotl
Label: Beta3Tubulin labeled with mCherry and CNP labeled with GFP
Instrument: Zeiss AxioZoom V16
Scale: 3cmx1cm
Image or sample processing steps:
Life imaging of a transgenic axolotl. Axolotl anesthetized and mounted to face the objective lens. A stack was recorded and and MIP was created by using the “extended depth of focus” tool from the ZenBlue software.

Little monsters from the deep – Marko Pende

MDIBL Light Microscopy Facility at MDIBL

Little monsters from the deep

Sample: Two Axolotl tissue cleared with the DeepClear method
Label: Neurons labelled with Beta3Tubulin in Alexa568 and Schwann cells labeled with MBP in Alexa488
Instrument: Zeiss AxioZoom V16, Stereoscope, Zeiss Plan Z 1.0x/0.25 FWD 60mm, Darkfield and Fluorescent image
Scale: 3cmx1.5cm
Image or sample processing steps:
Samples were mounted in a tissue clearing medium (RI:1.45) in close vicinity to each other facing the objective lens. A z-stack was acquired and a MIP was generated using the “extended depth of focus” tool in the ZenBlue software

A-Maze_ing_brain – Frederic Bonnet

MDIBL Light Microscopy Facility at MDI Biological Laboratory

Brain is a complex organ that requires a complex network of blood vessels to works properly.

Sample: Zebrafish embryo
Label: Brain vasculature labeled with flk-GFP
Instrument: Nikon Point scanning C2plus on an inverted Nikon Ti-Eclipse with an Plan Apo λ 10x/0.45NA objective
Scale: this image represents 586*586 um
Image or sample processing steps:
The image is a maximum intensity projection. Colors indicate the depth with blue for the surface (0 um) and yellow bottom (182 um)

The Stealth cell – Frederic Bonnet

MDIBL Light Microscopy Facility at MDI Biological Laboratory

An isolated cell shaped like a Stealth aircraft seeing from the front. The blue area in the center represent the window from the cockpit, then both wing on both sides

Sample: 3T3 cell
Label: actin cytoskeleton labeled with Alexa Fluor 568 Phalloidin
Instrument: Zeiss Point scanning confocal LSM980 on an upright Zeiss Axio Examiner stand. Image taken with a Zeiss plan Apo 63X/1.40 oil objective at zoom 1.7 with airy scan 2
Scale: 77.99um X77.99um
Image or sample processing steps:
maximum intensity projection color coded with the rainbow turbo LUT

Eyes of Ibad – Frederic Bonnet

MDIBL Light Microscopy Facility at MDI Biological Laboratory

Doing biology is always paying attention to detail. Having a cell adopting a eye shape is original and rare. The cell is like watching what we are doing.

Sample: Pig kidney cells
Label: Nuclei labeled with DAPI, Actine labeled with phalloidin and Vesicules labeled with GFP
Instrument: Zeiss Imager M2 with apotome with a Zeiss plan Apo 63x/1.40 oil objective
Image or sample processing steps:
Max intensity projection and color coded to match DUNE colors

Demo day fun – Holly C. Gibbs

Microscopy and Imaging Center at Texas A&M University

When you need an interesting demo sample in a pinch, look no further than under your window frames. Arachnids lay eggs in readily identifiable sacs in which spiderlings develop. This sample, not yet ready to hatch, is still fairly transparent, and has easily identifiable structures (8 legs!) visible by excitation of intrinsic fluorescence.

Sample: Triangulate cobweb spider (Steatoda triangulosa)
Label: Autofluorescence at 488 nm and 561 nm excitation collected at 500-550 nm and 575-625 nm
Instrument: Light sheet fluorescence microscopy, 5X 0.16 NA detection lens, 5X 0.1 NA illumination lens
Scale: 1.2 x 1.2 mm
Image or sample processing steps:
Raw data, maximum intensity projection of z-stack in Imaris

Mandibles – Hannah Somers

MDIBL Light Microscopy Facility at MDI Biological Laboratory

Mandibles (the last thing you see on a summer night when you turn off your light in the tent)

Sample: Japanese beetle Popillia japonica, reflected light microscopy
Label: NA
Instrument: Relfected light, Nikon Ti-E Spinning Disk Confocal, 10x Objective
Scale: 10 cm x 5 cm
Image or sample processing steps:
The beetle was fixed prior to imaging, the z-stack taken was about 1000 um thick, every 10 um.

Microglial Tempest – Peter Favreau

University of Wisconsin – Madison Optical Imaging Core, Stem Pharm, Inc.

Organoids are comprised of different cell types grown together to model organ systems. Here, separate neural cell types are grown together to form a composite human neural organoid. Some cells are labeled to understand specific components of the organoid. Endothelial cells, labeled in green, form the building blocks for vascular growth. Microglia, labeled in magenta, are critical surveyors of the brain.

Sample: Human neural organoid
Label: Microglia labeled with Iba1 / Alexa-647; endothelial cells labeled with CD-31 / Alexa-488; Nuclei labeled with Hoechst-33342.
Instrument: Nikon A1R HD Upright Multi-Photon/Confocal microscope
Scale: 502 um x 502 um
Image or sample processing steps:
This is a maximum intensity image from a z-stack that was 175 um thick, with 0.375 um sectioning. The image was processed using Fiji.

Sunrise of Embryonic Development – Lorenzo Gallicchio

Cell Sciences Imaging Facility: Beckman Center at Stanford

One of the first and more crucial events of embryonic development is the subdivision of the embryo into regions. From each region specific organs and body structures will originate. Such division is orchestrated by the expression of specific genes, for example the gene snail (in cyan) will mark the mesoderm, from which bones and intestine will develop, while rhomboid (in yellow) is marking the neurogenic endoderm from which neuroblasts will originate. Once such subdivision is determined the embryo is fully oriented, from which comes the name “Sunrise of Embryonic Development”.

Sample: Drosophila melanogaster embryo
Label: Probes against snail mRNA (cyan, Cal Fluor 560), probes against rhomboid mRNA (yellow, cal Fluor 610), Cellular Membrane (white, Alexa 488), DNA (blue, DAPI)
Instrument: leica SP8 confocal microscope, 40X

Do not cross your own domain! – Lorenzo Gallicchio

Cell Sciences Imaging Facility: Beckman Center at Stanford

During Development, cells need to differentiate in order to give rise to specific organs and tissues. It is crucial that the right number of cells acquire a certain fate at the right moment. In this image, in cyan are marked cells that are expressing a gene (Senseless) that will turn them into sensory organs, while in magenta are marked cells that are expressing another gene (miR-9a), that in turn prevents them by acquiring the sensory organ fate. The two patterns are well defined and complementary, and perturbations in such organization will lead to an increase or decrease of sensory organs.

Sample: Drosophila Embryo
Label: probes against senseless mRNA (blue, Cal Fluor 560), Probes against pre-miR-9a (magenta, Cal Fluor 670), antibody against Senseless-GFP (cyan, Alexa 488), DNA (gray, DAPI)
Instrument: Leica SP8 confocal microscope, 40X

When? Where? and How much? – Lorenzo Gallicchio

Cell Sciences Imaging Facility: Beckman Center at Stanford

In the drosophila testicle it is possible to follow visually the process of differentiation from the tiny stem cells located at the tip of the organ until the long sperm cells. At different stages of this differentiation process, genes are generally transcribed at different rates, and while stem cells and mature sperm do not have many mRNA molecules, big spermatocytes and elongating spermatids have many.

Sample: Drosophila melanogaster male gonad
Label: probes against polyA mRNA (white, Cal Fluor 610), DNA (blue, DAPI)
Instrument: Leica SP8 Confocal, 40X
Image or sample processing steps:
the image is the result of 9 different images merged together and acquired via the function tilescan.

Lipid string coiled around mouse sperm flagellum – Alejandro Linares

Laboratorio Nacional de Microscopía Avanzada (UNAM, MX) at @alinaresbio

Sperm cells contain a mitochondrial sheath in the upper portion of their flagellum. This region, known as the midpiece, is responsible for producing the energy required by the cell to reach the egg and ultimately achieve fertilization. The plasma membrane surrounding this region showed a fascinating (although mysterious) fluorescent labeling pattern, similar to a perfectly coiled string. This unusual event was accompanied by the fact that the cell was still alive, which means that this amazing observation might be physiologically relevant to the cell function.

Sample: Mouse sperm
Label: Plasma membrane labeled with BODIPY-Ganglioside GM1
Instrument: ONI Nanoimager-S Microscope in TIRF mode, 100X 1.49 NA
Scale: This image represents 35 µm^2
Image or sample processing steps:
This is super-resolution image reconstruction of a snapshot of a single mouse sperm cell. The algorithm used is MSSR (ImageJ plugin), with parameters: AMP = 2, Order = 0, FWHM = 2.5, Interpolation = Bicubic, Meshing Minimization = Enabled.

Mouse sperm ghosts – Alejandro Linares

Laboratorio Nacional de Microscopía Avanzada (UNAM – MX) at National Autonomous University of Mexico

These mouse sperm cells decided to prepare early for this years’ Halloween by putting on a mesmerizing ghostly costume, delivered by fluorescently labeling the plasma membranes of these cells.

Sample: Mouse sperm
Label: Plasma membrane with BODIPY – Ganglioside GM1
Instrument: ONI Nanoimager-S microscope in TIRF mode, 100X 1.49 NA
Scale: This image represents 56 x 44.5 µm
Image or sample processing steps:
This is a super-resolution image reconstruction comparison of two snapshots of mouse sperm cells. The algorithm used was MSSR (ImageJ plugin), with parameters: AMP = 2, Order = 0, FWHM = 2.5, Interpolation = Bicubic, Mesh minimization = enabled. Diffraction-limited images were combined with the super-resolved counterparts using ImageJ for comparison.

Muscle Head zebrafish – Romain Menard

MDIBL Light Microscopy Facility at MDIBL

This is skeletal muscles present in the head of a zebrafish Larvae

Sample: zebrafish
Label: Myosin labeled with F310
Instrument: Confocal

Motoneuron in the trunk of a zebrafish – Romain Menard

MDIBL Light microscopy Facility at MDIBL

Motoneurons projection in muscle tissues in the trunk of zebrafish Larvae 5dpf.

Sample: zebrafish
Label: Motoneuron labeled acetyl-tubulin
Instrument: Zeiss apotome

Motoneurons synapse in muscle tissues of a zebrafish trunk – Romain Menard

MDIBL Light microscopy Facility at MDIBL

This is a projection in skeletal muscle tissue of motoneuron synapses in the trunk of zebrafish.

Sample: zebrafish
Label: Synapse labeled with SV2
Instrument: Zeiss apotome

Going with the flow – Danya Alomar

Cell Biology and Image Acquisition Core Facility at University of Ottawa

This image shows a microsphere travelling through a large blood vessel of the mouse brain. Fluorescent microspheres were injected into the brain to induce micro-vessel occlusions. This models cerebral microinfarction, which is a form of “silent” stroke that is associated with vascular dementia and cognitive impairment. Microglia, the resident immune cells of the central nervous system (labeled green in this image), may play an important role in responding to microvascular injury caused by microinfarcts.

Sample: Sample taken from the thalamic region of the mouse brain. Animal underwent transcardial perfusion with phosphate-buffered saline and paraformaldehyde. Brain tissue was extracted and placed in sucrose solution.
Label: Blood vessels are labeled with Lycopersicon esculentum (Tomato) Lectin DyLight 594. Microglia is labeled with EGFP expressed in Tmem119-2A-EGFP mouse line.
Instrument: Fluorescence imaging on Zeiss Axio Imager M2 equipped with ApoTome.2, objective 20x.
Scale: 0.227 um x 0.227 um x 0.520 um
Image or sample processing steps:
This is a z-stack obained from a tissue section that was 100um thick. 75 slices (38 um tissue thickness) were imaged with the Apotome camera in the EGFP and Texas Red channels. The image was deconconvoled before exporting. Maximum projection of intensities was obtained in FIJI.

Mitochondria Party – Caroline Halluin

MDIBL Light Microscopy Facility. at MDI Biological Laboratory

Living mitochondria inside zebrafish muscle fibers

Sample: zebrafish 6dpf, live imaging
Label: Live imaging of mitochondria in trunk skeletal muscle fibers in a ubi:mito-mCherry transgenic line at 6 dpf.
Instrument: Zeiss LSM 980

Muscle Engine – Caroline Halluin

MDIBL Light Microscopy Facility. at MDI Biological Laboratory

Zebrafish muscle fibers and mitochondria, the powerhouse of the cells, providing energy for muscle contractions

Sample: zebrafish 6dpf, fixed, immunostaining
Label: Mitochondria (red) in slow (A41025 – magenta) and fast (F310 – green) trunk skeletal muscle fibers in a ubi:mito-mCherry transgenic zebrafish line at 6 dpf.
Instrument: Zeiss LSM 980
Image or sample processing steps:

Muscle tree – Caroline Halluin

MDIBL Light Microscopy Facility at MDI Biological Laboratory

Myosin expression in zebrafish muscle tissue

Sample: zebrafish 6dpf, fixed, immunostaining
Label: Immunostaining using the F310 antibody reveals myosin expression in fast trunk skeletal muscle 6 days old zebrafish larvae
Instrument: Zeiss LSM 980

Straight fiber – Pablo J. Sáez

UKE Microscopy Imaging Facility (UMIF) under the DFG Research Infrastructure Portal: RI_00489 at Cell Communication and Migration Laboratory, Institute of Biochemistry and Molecular Cell Biology, UKE

The image shows the actin cytoskeleton of a migrating cell.

Sample: Human umbilical vein endothelial cell (HUVEC)
Label: Actin labeled with expression of F-tractin GFP
Instrument: Widefield microscopy, Leica DMi8, Obj. HC PL FLUOTAR (40x/0.8)
Scale: 170 um x 100 um
Image or sample processing steps:
The image is a single plane, single time point of a F-Tractin GFP-expressing migrating cell. The image was subjected to background subtraction, and the Red Hot LUT was applied.

C. elegance – Michael Shribak

Label-free live cell imaging of C.elegans with quantitative orientation-independent differential interference contrast microscope
scale bar 100µm

Sample: organism
Label: label-free
Instrument: OI-DIC, Olympus IX81, 40x/0.6NA objective lens
Scale: 223 um x 372um

Cobblestones of the skeletal muscle – Elham Javed

Center for Translational Medicine at Thomas Jefferson University

Actin (green), fibroblasts (blue) and mitochondria (magenta) form the foundation of a skeletal muscle

Sample: Murine skeletal muscle was fixed and cryo sectioned, 10 um sections were stained.
Label: Actin was labelled using phalloidin 488. Fibroblasts were labelled using Vim -Cy3 (shown in blue). Mitochondria labelled with VDAC -647 (shown in magenta)
Instrument: Nikon A1R, using a 20x water objective. Image was acquired using a 1.2 zoom and as a z-stack of 22 slices with 0.65 um step size
Image or sample processing steps:
Image was processed using 2D deconvolution in NIS Elements

Eye of Sauron – Elham Javed

Center for Translational Medicine at Thomas Jefferson University

Axial section of murine brain showing the different stages of neuronal development

Sample: Murine brain tissue
Label: Vim (red), NeuN (green) and GFAP (yellow)
Instrument: Nikon A1R
Image or sample processing steps:
Image was acquired using 60x objective as a Z-stack of 8 slices using a step size of 1 um

Shape of my heart – Elham Javed

Center for Translational Medicine at Thomas Jefferson University

Cellular structure of the heart.

Sample: 10 um cryo section of a murine heart
Label: Desmin labelled with Alexa 488, Vimentin labeled with Cy3 (shown in magenta), Tropomyosin T labeled with Alexa 647 (shown in blue). DAPI (shown in white)
Instrument: Nikon AIR
Image or sample processing steps:
Image acquired using 40x Oil objective as Z-stack of 15 slices with a step size of 0.5 um. Image was processed with 2D Deconvolution using NIS Elements.

UFO, unidentified flying object – Aissette Baanannou

MDIBL Light Microscopy Facility at MDIBL

UFO, unidentified flying object

Sample: Zebrafish retina
Label: Green and red are cells in division
Instrument: Nikon C2+ point scanning with an inverted Nikon Ti-E and with a Nikon Plan Apo 10X/0.45 dry
Scale: 358*358 microns
Image or sample processing steps:
Image denoised using Nikon denoise.ai

Sun rise – Aissette Baanannou

MDIBL Light Microscopy Facility at MDIBL

Sun rise behind a planet

Sample: Zebrafish olfactory epithelium
Label: Olfactory sensory cells are labeled in red and neurons in green
Instrument: Zeiss LSM 980
Scale: 358*358 microns
Image or sample processing steps:

All paths meet – Aissette Baanannou

MDIBL Light Microscopy Facility at MDIBL

All paths meet for a good sight

Sample: zebrafish retina
Label: Blood vessels are labeled in red and photoreceptors and optic nerve are labeled in green
Instrument: Nikon C2+ point scanning
Scale: 358*358 microns

Plastic Wrap Cell – Tejeshwar Rao

University of Alabama at Birmingham

This represents an artistic presentation of a cell that might be wrapped using plastic wrap

Sample: Cos-7 Cell (African Green Monkey Kidney Fibroblast)
Label: Actin cytoskeleton and membrane
Instrument: Total Internal Reflection Fluorescence Microscopy, Nikon Ti2, 60X, 1.49
Scale:
Image or sample processing steps:
Image processed using Fiji

Crayon Cell – Tejeshwar Rao

University of Alabama at Birmingham

Crayon Cell

Sample: Cos-7 Cell (African Green Monkey Kidney Fibroblast)
Label: Actin cytoskeleton in orange (488 nm), paxillin in blue (647 nm), cellular tension in green (561 nm)
Instrument: Total Internal Reflection Fluorescence Microscopy, Nikon Ti2, 60X, 1.49

A broken heart – Josué David Hernández Varela

Instituto Politécnico Nacional

A broken heart

Sample: Fibroblast cell from inmortalized human (CHON) grow in stain steel support for protesis purposes
Label: Actin cytoskeleton and focal adhesion staining kit
Instrument: Spectral channels with LSM 880 coupled with a Elyra Ps.1 (Zeiss), 63X/1.4 DIC oil immersion objetive
Scale: This images represent a 300 x 300um
Image or sample processing steps:
This is a crop from a expanded image

Viral Neuroinvasion – Annika Lea Schulz

Life Sciences Institute Imaging – Advanced Bioimaging Core at University of British Columbia

Organoid-Based COVID-19 Research: Identifying Potential Therapeutic Agents Against SARS-CoV-2 Using Lab-Grown Mini Brains. Organoid models are emerging as valuable tools to investigate COVID-19. The neuro-serine protease inhibitor (neuroserpin) Spn4A, is an antiviral biologic discovered in the Jean Lab (Department of Microbiology & Immunology, Life Sciences Institute, University of British Columbia) currently being explored as a SARS-CoV-2 entry inhibitor. Here, the biological and antiviral activities of adenoviral-vector mediated Spn4A expression (Spn4A, in cyan) are investigated in human induced pluripotent stem cell-derived dorsal forebrain organoids. In addition, immunofluorescence staining indicates the presence of known respiratory tract cell entry factors of human coronaviruses (angiotensin converting enzyme-2 (ACE-2) in red, and transmembrane serine protease 2 (TMPRSS2) in green).

Sample: Human induced pluripotent stem cell-derived dorsal forebrain organoid with adenoviral-vector mediated expression of neuro-serine protease inhibitor (neuroserpin) Spn4A.
Label: The cell nuclei were stained with the fluorescent dye Hoechst – Pseudocolour: blue. The angiotensin converting enzyme-2 (ACE-2) was labelled with primary antibody: anti-ACE2 antibody (ab15348) & secondary antibody: gt-anti-rb Alexa647 – Pseudocolour: red. Transmembrane Serine Protease 2 (TMPRSS2) was labelled with primary antibody: anti-TMPRSS2 antibody (clone P5H9-A3, Cat. No. MABF2158) & secondary antibody: gt-anti-ms Alexa488 – Pseudocolour: green. The neuro-serine protease inhibitor was expressed using an adenoviral vector labelled with red fluorescent protein – Pseudocolour: cyan.
Instrument: Leica SP5 Inverted Confocal Microscope; 63X silicon oil immersion; tile scan protocol

Cerebral Nebula – Annika Lea Schulz

Life Sciences Institute Imaging – Advanced Bioimaging Core at University of British Columbia

Organoid-Based COVID-19 Research: Using Lab-Grown Mini Brains To Investigate Coronavirus Infection Of The Central Nervous System. Organoid models are proving their worth in verifying the safety and efficacy of antiviral therapies, as these in vitro models can exploit a priori knowledge of virus biology and allow for the testing of preclinical drugs as well as the discovery of new drugs. Shown here is a human pluripotent stem cell-derived dorsal forebrain organoid infected with human coronavirus 229E (HCoV-229E), which is one of the viruses responsible for the common cold. Immunofluorescence labelling indicates the presence of the known HCoV-229E cell entry receptor human aminopeptidase N (hAPN, in red) in the cerebral organoid. In addition, staining for double-stranded RNA (dsRNA, in green) is shown to visualize viral replication.

Sample: Human induced pluripotent stem cell-derived ventral forebrain organoid infected with human coronavirus 229E. Clearing was performed following immunofluorescence staining using fructose-glycerol clearing reagent.
Label: The cell nuclei (DNA) were stained with the fluorescent dye Hoechst – Pseudocolour: blue. CD13/ human aminopeptidase N (hAPN) was labelled with primary antibody: Anti-CD13 /hAPN: (GTX64331) – Genetex // Secondary: gt-anti-rb Alexa555 – Pseudocolour: red. Double-stranded RNA (dsRNA) was labelled with primary antibody: ms-J2-dsRNA (Scicons English and Scientific Consulting) & secondary antibody: gt-anti-ms Alexa488 – Pseudocolour: green.
Instrument: Leica SP5 Inverted Confocal Microscope; 63X silicon oil immersion; tile scan protocol

connection – Nana Kamei

connection – Nana Kamei MDIBL Light Microscopy Facility at MDI Biological Laboratory

A new nephron in regenerating zebrafish kidney has invaded and connected with an old nephron.

Sample: transgenic adult zebrafish regenerating kidney, 4%PFA fixed, mounted in 50%PBS/glycerol
Label: lhx1a:gfp+ distal new nephron labeled with AF488, actin cytoskeleton labeled with AF546-phalloidin, nuclei labeled with Hoechst 33342
Instrument: confocal microscopy, Zeiss 980 with Airyscan 2, 63x/1.4 oil objective
Scale: 78.38×78.38 microns
Image or sample processing steps:
this is a confocal slice 0.15 microns thick, processed with airyscan2

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RRID: SCR_024409

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