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Image Contest 2024

We are so thankful for all our 2024 Image Contest Contributors!!!

The Top three images are:

Sweet Embryo – Travis D. Carney
Quinoa Pollen Grain – Anastasiya Klebanovych
I love lamp – Hannah Somers

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

The People’s Choice Award is: Lysosome’s highway – Julien Cicero

Honorable Mentions (alpha order)

Hellebore pollen grain – Anastasiya Klebanovych
Burned eyes – José Manuel Martínez López
Arising Nervous System – Marko Pende
Gnat viewed from above – Laurel Schuck

Quinoa pollen grain – Anastasiya Klebanovych

Hellebore pollen grain – Anastasiya Klebanovych

Burned eyes – José Manuel Martínez López

Margarita STEM – José Manuel Martínez López

The universe and the nebulae that live within us – Brandon Antonio Segura Torres & Priscilla Vieto Bonilla

Microscopic nebulae. – Brandon Antonio Segura Torres & Priscilla Vieto Bonilla

Vincent Van Gogh under the microscope – Brandon Antonio Segura Torres & Priscilla Vieto Bonilla

Mosquito larva – Priscilla Vieto Bonilla & Brandon Antonio Segura Torres

Copepod – Priscilla Vieto Bonila & Brandon Antonio Segura Torres

Daphnia – Priscilla Vieto Bonilla & Brandon Antonio Segura Torres

Microtubule dance – Carla Maria Jaramillo Restrepo

A tale of two networks – Carla Maria Jaramillo Restrepo

Armonic duets – Carla Maria Jaramillo Restrepo

Angular Diffusion of Mesenchymal Stem Cells – Aaron Dhanda

Thought _the_Immaterium – Frederic Bonnet

2024 Cicada Brood XIX – cicada wing – Jenny Schafer

Inside the head of a zebrafish – Caroline Halluin

Viruses infecting bacteria – Jyot Antani, Ph.D.

Organoid Dynamics in 3D – Monica E. Brown

Vascular and Pericyte Dance in Zebrafish Retina – Anindita Neog

Garter snake shed skin autofluorescence – Laurel Schuck

Early blood vessel formation – Hannah Somers

Mycelium : Food Fungus – Ramdattu Santhapur and Dhimitraq Nikolla

Threading Chicken: Exploring the Inner Fiber Web – Ramdattu Santhapur and Dhimitraq Nikolla

Haunted Fibers: The Ghostly Secrets of Chicken Microstructures – Ramdattu Santhapur and Dhimitraq Nikolla

ATK Heart – Hannah Somers and Anastasia Paulmann

Flower by Drosophila – Andrés Gutiérrez-García & Mariana Marques-Reis

Intricate Interactions: The Dance of Microtubules and Actin – Wen Lu and Vladimir I. Gelfand

Space dish – Caramai Kamei (Image) and Omodasola Adekeye (staining)

Sweet Embryo – Travis D. Carney

Mount Desert Island Biological Laboratory
MDIBL Light Microscopy Facility

Fruit fly embryo

Sample: Drosophila embryo mounted in Vectashield
Label: Mouse anti-FasIII primary antibody + Alexa 647-conjugated secondary antibody.
Instrument: Zeiss LSM 980
Scale: Embryo is about 500 x 280 microns in size.
Image or Sample Processing Steps: Color-coded projection: different focal planes are in different colors.

Quinoa pollen grain – Anastasiya Klebanovych

Donald Danforth Plant Science Center
Advanced Bioimaging Laboratory

This is the maximum intensity projection of intensities from a 12.4-um z-stack. The image was processed in Zen (Black edition) software using SIM2 processing package.

Sample: This fluorescence image shows a quinoa pollen grain, captured using advanced super-resolution microscopy techniques. The vibrant colors come from special fluorescent dyes: propidium iodide and calcofluor white. Quinoa is a remarkable crop, known for its nutritional value and resilience in various growing conditions making it excellent model in plant research.
Label: Pollen grain from quinoa plant (wild type). Sample was fixed, rinsed and processed in ascending concentrations of TDE (2,2′-thiodiethanol) to equalize refractive index across the sample components (exine, intine) and to clear the sample. Imaging done using Zeiss Elyra 7, 63x oil objective in Lattice SIM mode. Images reconstruction was done using Zen black software and SIM2 processing package.
Instrument: Calcofluor white, a blue fluorescent dye, binds to polysaccharides found in cellulose, highlighting the plant cell walls. Propidium iodide, a red fluorescent dye, binds to pectins, another essential cell wall component.
Scale: Zeiss Elyra 7, Lattice SIM, Plan Apochromat 63x/ NA 1.4 oil DIC M27 objective, Z stack
Image or Sample Processing Steps: 1250×1226 pixels, 39 x 38 microns, 300 dpi

I love lamp – Hannah Somers

Mount Desert Island Biological Laboratory
MDIBL Light Microscopy Facility

A beautiful moth imaged using the Nikon T-ie spinning disk confocal.

Sample: A moth found in a spider web
Label: Autofluorscence was imaged.
Instrument: Reflected light, Nikon Ti-E YokoGawa CSU-W1 / Nikon C2+ using the 4x Objective 0.2 NA
Scale: 6 mm x 4 mm
Image or Sample Processing Steps: A z-stack and tiling were used to acquire the image, and finally a max intensity projection was created using the Nikon software. LUT were applied on Fiji.

Lysosome’s highway – Julien Cicero

University of California San Diego

In neurodegenerative disease research, fibroblasts extracted from a patient’s skin biopsy can be reprogrammed into induced pluripotent stem cells (iPSCs). These iPSCs are then used to generate neurons for study models. The image depicts lysosomes traveling along the axonal network of iPSC-derived motor neurons. Monitoring lysosome mobility and dynamics provides valuable insights, as their behavior can serve as a marker for neurodegenerative conditions.

Sample: human iPSC-derived motor neuron
Label: Axon membranes are stained with lipidic marker Bodipy-488 and lysosome with LysoTracker Deep Red
Instrument: LSM980 with Airyscan, obj 60X/1.4
Scale: 78 x 38 microns
Image or Sample Processing Steps: Magenta and Cyan LUT were apply in FIJI

Hellebore pollen grain – Anastasiya Klebanovych

Donald Danforth Plant Science Center
Advanced Bioimaging Laboratory

This image showcases a pollen grain from a hellebore plant, a perennial flowering species from the Ranunculaceae family, known for its beautiful ornamental qualities. The vibrant colors in the image result from autofluorescence, a natural phenomenon where certain components of the pollen grain, such as the exine (the outer layer) and cell wall, emit light when exposed to specific excitation light. This autofluorescence allows us to see intricate details and structures within the pollen grain that are otherwise invisible to the naked eye.

Sample: Pollen grain from Hellebore plant. Sample was collected and fixed, rinsed and processed in ascending concentrations of TDE (2,2′-thiodiethanol) to ensure clearing as well as refractive index equalization across pollen components.
Label: Autofluorescence of pollen wall components
Instrument: Lattice SIM, Zeiss Elyra 7, C-Apochromat 40x/ NA1.2 W CorrFCS
Scale: 3000×3000 pixels, 150 x 150 microns, 300 dpi
Image or Sample Processing Steps: This composite image was made using single 2D maximum intensity projections of 1024×1024 pixels featuring the same field of view but different pseudo-colors (lookup tables). Maximum intensity projections were done from a 32-um z-stack with 0.2 micron z-step interval. Image registration and processing was completed using Zen (Black edition) software with SIM2 processing package from Zeiss.

Burned eyes – José Manuel Martínez López

Química Tech

These are the eyes in auto-fluorescence of a scorpion (Centruroides exilicauda) after several hours under the microscope.

Sample: Centruroides exilicauda
Label: Auto-fluorescence
Instrument: Fluorescence microscopy, Microscope AxioImager, Objetive 10x/0.3, Filter set Ex 365/30, BS 405, Em 415LP
Scale: 1.4 x 1.05 mm
Image or Sample Processing Steps: Focus stacking from 110 images.

Arising Nervous System – Marko Pende

MDIBL

Tissue cleared and antibody labelled E12.5 mouse embryo labeled with anti-NF and SOX10 showing Peripheral glia and Neurofilament

Sample: Dehydration based tissue-clearing of an E12.5 mouse embryo
Label: anti-NF labeled with Alexa 568 and SOX10 labelled with Alexa 647 showing Peripheral glia and Neurofilament
Instrument: Light-sheet microscope, Custom build Ultramicroscope recorded with a Olympus 4x NA 0.28 corrected for a refractive index of 1.56
Scale: 4mm x 3mm
Image or Sample Processing Steps: Maximum intensity projection from two 2.3mm thick z-Stacks

Gnat viewed from above – Laurel Schuck

University of Minnesota
University Imaging Centers

This is a high resolution fluorescence image of the wings and back of a gnat. No labeling was done; the gnat went straight from the windowsill of the lab to on a slide under a microscope.

Sample: gnat (presumably? found on lab windowsill)
Label: no labeling, just autofluorescence
Instrument: point scanning confocal microscopy, Nikon A1R HD multiphoton, 25x/1.1NA (water immersion)
Scale: this image represents 1.53mm x 1.53mm
Image or Sample Processing Steps: This is a maximum intensity projection of a z stack that was 480um thick with 8um sectioning. Each z plane represents a single field of view (no tiling). For each plane, 405nm and 640nm laser excitation channels were collected simultaneously followed by collection of the 488nm channel. Prior to MIP assembly, Nikon Elements denoise.ai was used to denoise the images. Brightness adjustments were made for each channel.

Tail – José Manuel Martínez López

Química Tech

This is the tale of a scorpion observed in auto-fluorescence.

Sample: Centruroides exilicauda
Label: Auto-fluorescen
Instrument: Fluorescence microscopy, AxioImager.M2, Objective 2.5x/0.06 Filter dual band Ex 480/20-560/20, BS 500,580, Em 525/20,605/20
Scale: 5.58 x 4.21 mm
Image or Sample Processing Steps: Focus stacking from 26 widefield images

Margarita STEM – José Manuel Martínez López

Química Tech

This is the stem of a daisy flower observed in autofluorescence. The chlorophyll is fluorescing in red.

Sample: Daisy flower
Label: Autofluorescence
Instrument: Fluorescence microscopy, AxioImager M2, Objetive 2.5x/0.06 Filter Ex 420/40, BS 460, Em 470LP
Scale: 5.58 x 4.21 mm
Image or Sample Processing Steps: Focus stacking from 34 widefield images

Whole Mouse Brain – Dr. Edna Hardeman

University of Sydney
Bio-Imaging Facility (BMIF)

PEGASOS cleared whole mouse brain, Life-act EGFP transgenic actin filaments staining, tdTomato transgenic MARCKS plasma membrane staining.

Sample: Mouse brain
Label:
Instrument: Alpha3 Light Sheet Microscope by PhaseView
Scale:
Image or Sample Processing Steps:

Mouse brain – Boaz Mohar

Janelia Research Campus

Imaging of Mouse brain by PhaseView Alpha3 Light Sheet Microscope

Sample: Mouse brain
Label:
Instrument: Alpha 3 Light Sheet Microscope by PhaseView
Scale:
Image or Sample Processing Steps:

The universe and the nebulae that live within us – Brandon Antonio Segura Torres & Priscilla Vieto Bonilla

Universidad Nacional del Comahue

Section cut of stomach tissue stained with hematoxylin and eosin, at 800x magnification, dark field, focus tracking with stacking of 32 images.

Sample: Section cut of stomach tissue stained with hematoxylin and eosin, at 800x magnification, dark field, focus tracking with stacking of 32 images.
Label: Section cut of stomach tissue stained with hematoxylin and eosin
Instrument: elestron Labs Cb1000cf Binocular Microscope Immersion, 800x magnification, Nikon D5300 camera.
Scale: x800
Image or Sample Processing Steps: Section cut of stomach tissue stained with hematoxylin and eosin, at 800x magnification, dark field, focus tracking with stacking of 32 images.

Microscopic nebulae. – Brandon Antonio Segura Torres & Priscilla Vieto Bonilla

Univesidad Nacional del Comahue

Section cut of stomach tissue stained with hematoxylin and eosin, at 800x magnification, dark field, focus tracking with stacking of 43 images.

Sample: Section cut of stomach tissue stained with hematoxylin and eosin, at 800x magnification, dark field, focus tracking with stacking of 43 images.
Label: Section cut of stomach tissue stained with hematoxylin and eosin, at 800x magnification, dark field, focus tracking with stacking of 43 images.
Instrument: Celestron Labs Cb1000cf Binocular Microscope Immersion, 800x magnification, Nikon D5300 camera.
Scale: x800
Image or Sample Processing Steps:
Section cut of stomach tissue stained with hematoxylin and eosin, at 800x magnification, dark field, focus tracking with stacking of 43 images.

Vincent Van Gogh under the microscope – Brandon Antonio Segura Torres & Priscilla Vieto Bonilla

Universidad Nacional del Comahue

Stratified squamous tissue stained with hematoxylin and eosin, at 400x magnification, dark field, focus tracking with stacking of 28 images.

Sample: Stratified squamous tissue stained with hematoxylin and eosin, at 400x magnification, dark field, focus tracking with stacking of 28 images.
Label: Stratified squamous tissue stained with hematoxylin and eosin, at 400x magnification, dark field, focus tracking with stacking of 28 images.
Instrument: Celestron Labs Cb1000cf Binocular Microscope Immersion, 800x magnification, Nikon D5300 camera.
Scale: x400
Image or Sample Processing Steps:
Stratified squamous tissue stained with hematoxylin and eosin, at 400x magnification, dark field, focus tracking with stacking of 28 images.

Mosquito larva – Priscilla Vieto Bonilla & Brandon Antonio Segura Torres

Universidad Nacional del Comahue

Mosquito larva viewed in dark field and birefringent polarized light, image stacking and focus tracking of 120 images in 5 different sections.

Sample: Mosquito larva viewed in dark field and birefringent polarized light, image stacking and focus tracking of 120 images in 5 different sections.
Label: Mosquito larva viewed in dark field and birefringent polarized light, image stacking and focus tracking of 120 images in 5 different sections
Instrument: Celestron Labs Cb1000cf Binocular Microscope Immersion, 25x magnification, Nikon D5300 camera.
Scale: 25x
Image or Sample Processing Steps: Mosquito larva viewed in dark field and birefringent polarized light, image stacking and focus tracking of 120 images in 5 different sections

Copepod – Priscilla Vieto Bonila & Brandon Antonio Segura Torres

Universidad Nacional del Comahue

Copepod viewed in dark field with birefringent polarized light, with stacking of 63 images in two different sections and focus tracking, at 400x magnification.

Sample: Copepod viewed in dark field with birefringent polarized light, with stacking of 63 images in two different sections and focus tracking, at 400x magnification.
Label: Copepod viewed in dark field with birefringent polarized light, with stacking of 63 images in two different sections and focus tracking, at 400x magnification.
Instrument: Celestron Labs Cb1000cf Binocular Microscope Immersion, 800x magnification, Nikon D5300 camera.
Scale: 400x
Image or Sample Processing Steps: Copepod viewed in dark field with birefringent polarized light, with stacking of 63 images in two different sections and focus tracking, at 400x magnification.

Daphnia – Priscilla Vieto Bonilla & Brandon Antonio Segura Torres

Universidad Nacional del Comahue

Daphnia viewed in dark field with birefringent polarized light, with stacking of 53 images in two different sections and focus tracking, at 400x magnification.

Sample: Daphnia viewed in dark field with birefringent polarized light, with stacking of 53 images in two different sections and focus tracking, at 400x magnification.
Label: Daphnia viewed in dark field with birefringent polarized light, with stacking of 53 images in two different sections and focus tracking, at 400x magnification.
Instrument: Celestron Labs Cb1000cf Binocular Microscope Immersion, 800x magnification, Nikon D5300 camera.
Scale: 400x
Image or Sample Processing Steps: Daphnia viewed in dark field with birefringent polarized light, with stacking of 53 images in two different sections and focus tracking, at 400x magnification.

Golden fibers – Julien Cicero

University of California San Diego

In the context of neurodegenerative disease research, fibroblasts can be extracted from a patient’s skin biopsy. These fibroblasts are then reprogrammed into induced pluripotent stem cells (iPSCs) to generate neurons for study models. The image shows an axonal network from iPSC-derived motor neurons used for molecular screening.

Sample: human iPSC-derived motor neuron
Label: Actin cytoskeleton labeled with SPY620-actin
Instrument: LSM980 with Airyscan SR, obj 60x/1.4, oil
Scale: 78 x 38 microns
Image or Sample Processing Steps: Yellow hot LUT was apply in FIJI

Microtubule dance – Carla Maria Jaramillo Restrepo

Centro de Investigaciones en Óptica (CIO)

HeLa cells, showcasing the intricate network of microtubules. Acetylated-tubulin highlights stable microtubules in red, while tyrosinated-tubulin reveals dynamic microtubules in green. The contrast between the stable and dynamic components of the cytoskeleton is vividly captured through immunostaining.

Sample: HeLa cells fixed and immunostained for post-translational modifications of tubulin markers.
Label: Stable microtubules (green) are highlighted by immunostaining with anti-alpha tubulin antibody for acetylated-tubulin. Dynamic microtubules (red) are revealed by staining with anti-alpha tyrosinated antibody for tyrosinated-tubulin. Nuclei (blue) are stained with DAPI.
Instrument: Confocal microscopy Zeiss LSM-710-NLO, 63X objective
Scale: 15.1795 pixeles/micron
Image or Sample Processing Steps:

A tale of two networks – Carla Maria Jaramillo Restrepo

Centro de Investigaciones en Óptica (CIO)

Sample: Sample: HeLa cells fixed and immunostained for post-translational modifications of tubulin markers.
Label: Stable microtubules (green) are highlighted by immunostaining with anti-alpha tubulin antibody for acetylated-tubulin. Dynamic microtubules (red) are revealed by staining with anti-alpha tyrosinated antibody for tyrosinated-tubulin. Nuclei (blue) are stained with DAPI.
Instrument: Confocal microscopy Zeiss LSM-710-NLO, 63X objective
Scale: 15.1759 pixeles/micron
Image or Sample Processing Steps:

Armonic duets – Carla Maria Jaramillo Restrepo

Centro de Investigaciones en Óptica (CIO)

Sample: HeLa cells fixed and immunostained for post-translational modifications of tubulin markers.
Label: Stable microtubules (green) are highlighted by immunostaining with anti-alpha tubulin antibody for acetylated-tubulin. Dynamic microtubules (red) are revealed by staining with anti-alpha tyrosinated antibody for tyrosinated-tubulin. Nuclei (blue) are stained with DAPI
Instrument: Confocal microscopy Zeiss LSM-710-NLO, 63X objective
Scale: 15.1759 pixeles/micron
Image or Sample Processing Steps:

Angular Diffusion of Mesenchymal Stem Cells – Aaron Dhanda

STEMCELL Technologies

Mesenchymal stem cells (MSCs) are at the forefront of leading advances in medical applications such tissue regeneration. A key aspect of their regenerative potential is the ability of MSCs to migrate. This image shows a culture of MSCs labelled with an orange dye that binds to actin, while a blue dye (DAPI) lights up the cell nuclei. The orange dye helps us see the striking pointed shapes of the cells and the network of actin filament structures inside the cells that enables the cells to move.

Sample: Paraformaldehyde Fixed Human Mesenchymal Stem Cells
Label: F-actin with Alexa488-phalloidin (pseudo-colored with the orange hot LUT) and DAPI (blue)
Instrument: Leica SP8 confocal microscope equipped with a 63x/1.40 oil immersion objective
Scale:
Image or Sample Processing Steps: This image is a tile scanned image that was combined using the LASX software; the only adjustments made were a crop followed by an adjustment of color levels (min and max levels) using ImageJ.

Molten Gateways – Frederic Bonnet

MDIBL
MDIBL Light Microscopy Facility

This mesmerizing image reveals the glowing, fiery pathways within renal glomeruli – the intricate filtering units of the kidney. Brilliantly fluorescent probes highlight the complex architectures that form the molecular gatekeepers governing what can enter and exit the bloodstream. The vibrant blue hues delineate the delicate scaffolding of actin filaments, while the incandescent Fire shades illuminate the glycoproteins enmeshing the vasculature. This dazzling display imparts an ethereal, molten quality, capturing the dynamic processes constantly churning to maintain homeostasis and life itself.

Sample: Glomerulus from Mouse kidney section
Label: Alexa Fluor 488 wheat germ agglutinin (W-11261)and Alexa Fluor 568 phalloidin(A-12380)
Instrument: Zeiss LSM980 on an Axio Examiner Z1, equipped with a zeiss Plan-Apochromat 63x/1.4 Oil and airyscan 2
Scale: 78.4 microns by 78.4 microns
Image or Sample Processing Steps: Single Z plane processed with FIJI

Thought _the_Immaterium – Frederic Bonnet

MDIBL
MDIBL Light Microscopy Facility

In this imaginative interpretation, the vibrant microscopic textures evoke the metaphysical chaos of the Warhammer 40K universe’s immaterium – the extra dimensional realm of psychic energy and untamed psychic phenomena. The glowing red forms become embodiment of warp entities, spectral fires, and churning psychic currents unleashing visions of unrestrained creation and destruction. This aims to connect the visuals to the vivid cosmic, psychic, and magical aspects of the 40K lore in an abstract expressionist manner.

Sample: Glomerulus from Mouse kidney section
Label: Alexa Fluor 488 wheat germ agglutinin (W-11261),Alexa Fluor 568 phalloidin(A-12380) and DAPI nuclei
Instrument: Zeiss LSM980 on anAxio Examiner Z1, equipped with a zeiss Plan-Apochromat 63x/1.4 Oil and airyscan 2
Scale: 78.4X78.4 microns
Image or Sample Processing Steps: Single Z processed with FIJI

2024 Cicada Brood XIX – cicada wing – Jenny Schafer

Vanderbilt University
Vanderbilt Cell Imaging Shared Resource (CISR)

Cicada wing mounted on glass coverslip and imaged on a Zeiss LSM710 confocal with autofluorescence, 20x/0.8 NA objective, Z-stack of 130 microns, color-coded for Z-depth

Sample: Cicada wing on a glass coverslip, no preparation
Label: Autofluorescence at 561 nm excitation
Instrument: Zeiss LSM710 confocal with 20x/0.8 NA objective
Scale: 425 x 425 x 130 microns
Image or Sample Processing Steps: Maximum Intensity Projection of 130 microns color coded for Z-depth

Inside the head of a zebrafish – Caroline Halluin

MDI Biological Laboratory
MDIBL Light Microscopy Facility

This is the head of a zebrafish larva at 5 days old. Neurons, nerves, synapses and sight in the larval zebrafish: retinal ganglion neurons project the optic nerve into the fish’s brain, enabling synaptic connections and functions.

Sample: 5 days old zebrafish larva, fixed. Immunostaining
Label: acetyl tubulin labelled with Alexa555- sv2 labelled with Alexa647- islet2b-GFP labelled with Alexa488
Instrument: Zeiss LSM 980
Scale:
Image or Sample Processing Steps: this is a maximum projection from a z stack that was105um thick with 3um sectioning

Viruses infecting bacteria – Jyot Antani, Ph.D.

Yale University

Bacteriophages are viruses that infect and often kill bacteria. The bright spots on the image are viruses labeled with a fluorescent dye, infecting the rod-shaped bacterial cells.

Sample: Bacteriphage H6 (~120 nm), Pseudomonas aeruginosa cells (2 um-long rods)
Label: Viral genome labeled with SYTOX Orange.
Instrument: Epifluorescence microscopy, Nikon Ti-E, Objective: 100X, 1.40 NA
Scale: this image represents 134 μm x 134 μm
Image or Sample Processing Steps: The sample contained live bacteria placed on an agarose pad. Pre-labeled virus-particles were introduced two minutes before image acquisition. This single image was taken with epifluorescence modality (white light from Lumencor SOLA engine, common Texas Red ex/em filter set). The image was false-colored using the mpl-inferno lookup table and brightness was adjusted. No decovolution was applied to this image.

Beta Cancri (crab) – Frederic Bonnet

MDIBL
MDIBL Light Microscopy Facility

Crab found on the beach near MDIBL.

Sample: Crab
Label: autofluorescence
Instrument: Fluorescence with a Nikon Ti-E YokoGawa CSU-W1 equipped with a CFI Plan Apochromat Lambda D 4X/0.2
Scale: 10mm X 5.6mm
Image or Sample Processing Steps: 4 color image (405-488-568-647nm), Z stack and max intensity projection, processed with FIJI

Organoid Dynamics in 3D – Monica E. Brown

Vanderbilt University

Human tissue-derived colon organoid. Blue represents nuclei and green represents cells in the cell cycle.

Sample: human colon organoid. whole-mount immunostained and imaged with clearing using fructose-glycerol solution.
Label: Nuclei labeled with Hoechst (blue). Dividing cells labeled with MKI67 with Alexa488 (green).
Instrument: Whole-mount fluorescent imaging on a Nikon A1R laser scanning confocal with a 20x/
Scale: 246.7um x 246.7um
Image or Sample Processing Steps: Maximum projection of intensities from a 55um thick z-stack with 1um sections.

Vascular and Pericyte Dance in Zebrafish Retina – Anindita Neog

Mount Desert Island Biological Laboratory
MDIBL Light Microscopy Facility

This confocal microscopy image presents a detailed view of the retina in a 5-day post-fertilization (5dpf) zebrafish larva. The green staining highlights the developing hyaloid vasculature, while the magenta staining marks the pericytes, which are associated with the blood vessels. The image reveals the intricate and organized pattern of the retinal vasculature into a hemispherical basket, along with the pericytes that play a crucial role in stabilizing and supporting these blood vessels. This visual representation is essential for understanding retinal development and the interactions between vascular structures and pericytes in zebrafish, a common model organism in developmental biology studies.

Sample: Retina of zebrafish larvae (5 days post fertilization)
Label: Blood vessels (Fli1a:GFP) labelled with GFP and pericytes (pdgfrb:mcherry) labelled with mcherry
Instrument: Point scanning confocal unit (LSM 980, Carl Zeiss Microscopy, Germany) on a Zeiss Axio Examiner Z1 upright microscope stand (ref: 409000-9752-000, Carl Zeiss Microscopy, Germany). Objective: Zeiss Plan-Apochromat 20X, Immersion: Air, NA: 0.80 (ref: 420650-9902-00020x, Carl Zeiss Microscopy, Germany)
Scale:
Image or Sample Processing Steps:

Sweet brains – Travis D. Carney

Mount Desert Island Biological Laboratory
MDIBL Light Microscopy Facility

Neurons in the brain of a fruit fly larva.

Sample: Drosophila central nervous system; mounted in Vectashield
Label: Left: Mouse anti-FasII + Alexa 488-conjugated secondary antibody; right: membrane-bound RFP driven by OK107-Gal4, labeled with mouse anti-RFP and an Alexa-568-conjugated secondary antibody.
Instrument: Zeiss LSM 980
Scale: CNS is approximately 250-300 microns across.
Image or Sample Processing Steps: Images are color-coded projections, so different focal planes are in different colors.

Sweet testes – Travis D. Carney

Mount Desert Island Biological Laboratory
MDIBL Light Microscopy Facility

Drosophila male reproductive system: paired coiled testes, seminal vesicles, and accessory glands, and a single ejaculatory duct.

Sample: Drosophila male reproductive system mounted in Vectashield + DAPI
Label: DAPI (cyan); Rabbit anti-Vasa + 488-conjugated secondary antibody (green); mouse anti-Dlg + 568-conjugated secondary antibody (magenta)
Instrument: Zeiss LSM 980
Scale: each coiled testis is about 1.5 mm long
Image or Sample Processing Steps: This is a maximum-intensity projection; sample is about 45 microns thick, and images were taken at 5 micron step sizes.

Garter snake shed skin autofluorescence – Laurel Schuck

University of Minnesota
University Imaging Centers

This is a high resolution image of a piece of snakeskin shed by a garter snake. There was no labeling done; the shed skin went straight from outside on the ground to on a slide under a microscope.

Sample: garter snake shed skin
Label: no labeling, just autofluorescence
Instrument: Point scanning confocal microscopy, Nikon AX R, 40x/0.95 NA
Scale: this image represents 330um x 330um
Image or Sample Processing Steps: This is a maximum intensity projection of a z stack that was 34um thick with 2um optical sectioning. For each frame, 405nm and 561nm laser excitation channels were collected simultaneously, then 488nm and 640nm excitation channels were collected simultaneously. This represents a single field of view (no tiling). Nikon Elements denoise.ai was used for denoising every channel prior to assembling the MIP.

Early blood vessel formation – Hannah Somers

Mount Desert Island Biological Laboratory
MDIBL Light Microscopy Facility

Endothelial cells interwoven with Laminin

Sample: Renal Organoid
Label: Magenta is Laminin (mCherry) and yellow is CD31 (GFP)
Instrument: Fluorescent microscopy, Zeiss LSM 980 – Airyscan 2 / MP, 63x objective, 1.4 NA/oil
Scale: 200 um x 200 um
Image or Sample Processing Steps: A z-stack was used to create this maximum intensity projection image.

Wing of green lacewing – Laurel Schuck

University of Minnesota
University Imaging Centers

This is a high resolution fluorescence image of the wing of a green lacewing. Specifically, the area near where the wing would connect to the body. No labeling was done; the insect was found in the lab, then a slide of just one wing was imaged.

Sample: wing of green lacewing
Label: no labeling, just autofluorescence
Instrument: point scanning confocal microscopy, Nikon A1R HD multiphoton confocal, 25x/1.1 NA (water immersion)
Scale: this image represents 1mm x 0.905mm
Image or Sample Processing Steps: This is a maximum intensity projection of a z stack that was 240 um thick, with 8um initial sectioning. Each z plane represents 63 tiles in xy stitched together via blending (15% overlap) in Nikon Elements. Nikon Elements denoise.ai was used to denoise individual frames prior to stitching and MIP assembly. For each frame, 405nm and 640nm laser excitation channels were collected simultaneously followed by collection of the 488nm excitation channel. Brightness adjustments were made to each channel.

Mycelium : Food Fungus – Ramdattu Santhapur and Dhimitraq Nikolla

University of Massachusetts Amherst
Light Microscope Facility- Institute for Applied Life Sciences

Sample was frozen at -23C and then cut using Cryostat with a thickness of 100um.

Sample: Raw Mycelium (fermented).
Label: Proteins were stained with FITC (green) and fats with Nile Red( Red).
Instrument: Laser Scanning Confocal Microscope with DigitalSight10 CCD (Nikon AXR-NSPARC)
Scale: Plan Apo 10X 0.45-NA objective
Image or Sample Processing Steps: LUTs adjustments on Nikon NIS Elements

Threading Chicken: Exploring the Inner Fiber Web – Ramdattu Santhapur and Dhimitraq Nikolla

University of Massachusetts Amherst
Light Microscope Facility- Institute for Applied Life Sciences

An in-depth analysis of raw chicken samples to investigate the microstructures, fiber orientation, length, and area, offering critical insights into the structural properties of poultry proteins for improved product development and texture optimization.

Sample: Tissue was frozen at -23C and then cut using cryomicrotome with a thickness of 100um
Label: Proteins with FITC (green), Fibers with Nile red (Red)
Instrument: Laser Scanning Confocal Microscope with DigitalSight10 CCD (Nikon AXR-NSPARC)
Scale: Plan Apo 10X 0.45-NA objective
Image or Sample Processing Steps: LUTs adjustments on Nikon NIS Elements

Rainbow embryo – Tatyana Vetter

Nationwide Children’s Hospital
NCH Microscopy Core

3D volume snapshot of neurofilament in a cleared mouse embryo

Sample: Mouse embryo cleared with iDISCO protocol
Label: Neurofilament antibody-labeled with a Alexa Fluor 488
Instrument: Nikon AX R confocal on a Nikon Ti2-E stand, 4x Plan Apo Lambda Objective (0.2 NA), 2048×2048 resonant scan with 8x averaging, 1.9 um/pixel, 10.2 um z-step, 1.1 mm sample thickness
Scale: 3.04 mm x 3.90 mm LxW, 1.14 mm depth
Image or Sample Processing Steps: The z-stack was denoised, projected in 3D volume view, and depth-coded. The image is a snapshot of the 3D projection from the XY view plane

Haunted Fibers: The Ghostly Secrets of Chicken Microstructures – Ramdattu Santhapur and Dhimitraq Nikolla

University of Massachusetts Amherst
Light Microscope Facility- Institute for Applied Life Sciences

A commercially available chicken burger patty was frozen at -23°C and sectioned into 100 µm thin slices using a CryoMicrotome. This process facilitated the examination of fiber microstructures and the identification of additional ingredients present within the pre-prepared burger patty, enabling detailed structural and compositional analysis.

Sample: Food Sample (Chicken Burger Patty)
Label: Proteins were stained with FITC (green) and fats with Nile Red(Red).
Instrument: Laser Scanning Confocal Microscope with DigitalSight10 CCD (Nikon AXR-NSPARC), Plan Apo 10X 0.45-NA objective
Scale: 20µm
Image or Sample Processing Steps: LUTs adjustments on Nikon NIS Elements

ATK Heart – Hannah Somers and Anastasia Paulmann

Mount Desert Island Biological Laboratory
MDIBL Light Microscopy
Facility

Blood vessels labeled in an African Turquoise Killifish

Sample: A whole mount heart labeled for blood vessels
Label: The heart was labeled using CD31 antibody.
Instrument: Fluorescent microscopy, Zeiss LSM 980 – Airyscan 2 / MP, 10x objective 0.45 NA
Scale: 3 mm x 1.5 mm
Image or Sample Processing Steps: Z-stack and tiling were used to create a maximum intensity projection of the ATK heart.

Flower by Drosophila – Andrés Gutiérrez-García & Mariana Marques-Reis

Northwestern University & Champalimaud Foundation

Crop of Drosophila Egg Chambers. Original image was taken over 32h on a Zeiss LSM 880 Airyscan, using the Plan-Apochromat 20x 0.8NA objective. Tile Scan. Each frame had a resolution of 4096*4096 pixels. IMARIS 3D Reconstruction. Colors correspond to endogenous proteins fused fluorescent proteins (GFP and mCherry) and DAPI staining for nuclei marking.

Sample: Drosophila Egg Chambers. Bridge Mounting.
Label: DAPI staining. GFP and mCherry mark endogenous proteins fused with those fluorescence proteins.
Instrument: Zeiss LSM 880 Airyscan, using the Plan-Apochromat 20x 0.8NA objective. Tile Scan. Each frame had a resolution of 4096*4096 pixels. 3D Reconstruction of an 80 micras z-Stack.
Scale:
Image or Sample Processing Steps: Tile Scan (Stitched with ZEN Black). Each frame had a resolution of 4096*4096 pixels. 80 micras z-Stack (1 micron step). Contrasted in FIJI. 3D Reconstructed on IMARIS (Viewer). Original tile image had a final resolution (post stitching) of 8191 x 12287 pixels for an image file of 44.9GB.

Intricate Interactions: The Dance of Microtubules and Actin – Wen Lu and Vladimir I. Gelfand

Northwestern University Feinberg School of Medicine

The image displays the intricate internal cytoskeletal structure of a nurse cell within a Drosophila ovary. Microtubules are labeled yellow, and actin filaments are labeled in magenta.

Sample: Drosophila melanogaster, nurse cells, fixed with 8% EM-grade formaldehyde in 1XBRB80 +0.1% Triton X-100 buffer.
Label: Microtubules labeled with EMTB-3XGFP, and actin filaments stained with Rhodamine-conjugated phalloidin.
Instrument: DeepSIM by Crestoptics; 60X oil lens, NA 1.40
Scale: 88.89 um x 110.93 um
Image or Sample Processing Steps: The image is a maximum projection of intensities from a z-stack that was 6.9 um thick, with 0.3 um sectioning; the image was contrasted and sharpened in FIJI.

New Flow – Caramai Kamei

MDIBL
MDIBL Light Microscopy
Facility

A regenerating new nephron formed after kidney injury is starting to filter blood and shows tiny vesicles labeled with dextran compared to large vesicles in existing nephrons.

Sample: whole mount PFA fixed adult zebrafish kidney 10 days post gentamicin injury with EdU injected at 6dpi and dextran injected at 9dpi
Label: EdU+ proliferating nuclei labeled with AF647, vesicular uptake of 10Kdalton rhodamine dextran in proximal tubules, Hoechst 33342 in nuclei
Instrument: Zeiss LSM980 Plan-Apo 10x/0.45
Scale: 494 microns x 494 microns
Image or Sample Processing Steps: single confocal slice with color brightness/contrast adjusted in FIJI

Outflow – Caramai Kamei

MDIBL
MDIBL Light Microscopy
Facility

This image is taken at the cloaca where excretion from the gut and kidney are taking place

Sample: 6dpf live zebrafish
Label: cytoplasmic membranes and lipids labeled with Bodipy TR Methyl Ester
Instrument: Zeiss LSM980 Plan-Apo 10x/0.45
Scale: 494 x 494 microns
Image or Sample Processing Steps: single confocal slice

Space dish – Caramai Kamei (Image) and Omodasola Adekeye (staining)

MDIBL
MDIBL Light Microscopy
Facility

Neuromasts in the zebrafish lateral line contain hair cells that sense movement and water flow like a satellite dish receiving signals from space

Sample: 20 micron cryosection of 5dpf PFA fixed zebrafish with immunofluorescent staining for ptprq, zo-1, nuclei
Label: Hair cells of the neuromast are labeled with AF546 in magenta, tight junctions labeled with AF488 in green, nuclei labeled with Hoechst 33342
Instrument: Zeiss LSM 980 Plan-Apo (10x/0.45)
Scale: 122 x 122 microns
Image or Sample Processing Steps: Maximum projection of z-stack 14 slices thick taken at optimal intervals with zen software and airyscan2 deconvolution

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RRID: SCR_024409

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