Microscopy Image Competition

Ryszard Wimmer: Institut Curie

Cerebral Organoid Ventricular Zone fixed with 4% PFA stained for Centrosomes with gamma-Tubulin, SOX2 for Radial Glia Cells, Nesprin-2 (25265-1-AP) imaged on an AXR confocal microscope 100x with 4.0 Niquist Zoom., Slice of Human Fetal Brain at Gestation Week 17 showcasing the Subventricular zone where most of the neural stem cells of the brain are located (Basal Radial Glia cells). These elongated stem cells of the brain have individual processes expandind all the way to the pial surface. ​
Human Fetal Brain (GW17), 2% PFA slow O/N fixation. ​
Magenta - Vimentin, Sox2 - Yellow (66411-1-Ig), Cyan - DAPI. ​
Mosaic Imaging on a Nikon W1 Spinning Wide Microscope at low resolution 20x dry objective.

Wen Lu: Northwestern University

Drosophilalarval salivary glands, fixed in 4% PFA in 1× PBS. On the left, nuclei are stained with DAPI and the internal lumen is labeled with tdTomato-tagged human CD4 protein (grey). On the right, microtubules are stained with CoraLite 488–conjugated anti-β-tubulin antibody (Proteintech, Cat# CL488-66240). Multiplexed hyperstacks were generated with spectrum color coding applied to the z-stacks of DAPIand microtubule staining. Images were acquired using a 20× laser scanning confocal microscope with large image stitching.

Bhavin Shah: Medizinische Fakultät Mannheim, Universität Heidelberg

4%PFA fixed P21 Mouse Liver Tissue stained for Liver Endothelial cells and other markers. Lyve1 (in Cyan, R&D #AF2125); Stab2 (in Magenta, Peptide speciality Lab #M881-P15) and CD68 (in yellow, Proteintech 25747-1-AP). ​
The tissue wascut on a vibratome (100µm thick) and stained with the above mentioned antibodies. Due to large background from the Livertissue, the samples after staining were Tissue-Cleared using Organic Solvents and then the cleared Tissue was imaged using zeiss-LSM confocal 800 Mikroscope at 40x. The image is a Z-Projection of 15 Stacks (each Stack imaged at 1µm z-interval).

Francisco  Lázaro-Diéguez: Albert Einstein College of Medicine

Dedifferentiated rat liver cells, fixed with 100 % methanol at -20 °C, and stained to visualize microtubules (α-tubulin, white), β-actin (red) (66009-1-Ig), and nuclei (DAPI, green).

Lobke Mombeek: Hasselt University

An E16 mouse embryo (4% PFA fixed) was processed using the iDISCO+ tissue clearing protocol. ​Immunolabeling was carried out with antibodies against Phox2B - orange (AF4940) to identify enteric neural precursors and Tubb3 - purple (PRB-435P) to visualize neuronal fibers. ​Three-dimensional imaging was performed on a Zeiss Lightsheet 7 microscope equipped with an EC Plan-NEOFLUAR 5x/0.16 objective. ​Imaging was conducted by Dr. Margarita Tevosian.

Luisa Barleben: Charité - Universitätsmedizin Berlin

A Tale of Two Colonoids: Life and Death in 3D. Side by side, one human colonoid thrives while the other succumbs. The 4% PFA-fixed cells are stained to reveal epithelial barrier (green, CDH1, BD Biosciences, Cat. 610182), proliferation (magenta, Ki67, Invitrogen, Cat. 14-5698-80), apoptosis (red, Cleaved Caspase 3, Cell Signaling, Cat. 9661), and nuclei (DAPI) — capturing a dramatic contrast in a single 3D projection., Colonic organoid grown from murine intestinal epithelial cells fixed in 4% PFA. This flattened Z-projection of the 3D immunofluorescence image shows both Zonula occludens-1 (red, Proteintech, 21773-1-AP) as well as E-cadherin (green, BD Biosciences, 610181) - two pivotal barrier proteins necessary for crypt architecture. Nuclei are counterstained with DAPI in blue. These epithelial colonoids illuminate the spatial relationship between adherens and tight junctions in the 3D tissue.

Bruno Cisterna: Augusta University

Immunofluorescence micrograph of neurons differentiated from induced pluripotent stem cells (iPSCs) after 62 days of maturation. Cells fixed with 3% paraformaldehyde and 0.1% glutaraldehyde. The actin cytoskeleton is orange, microtubules in cyan, and nuclei in yellow. Imaging using a Nikon CSU-W1 SoRa spinning disk confocal microscope equipped with a Hamamatsu ORCA-Fusion BT camera and a 40x objective lens., Super-resolution spinning disk confocal image of a cath.a-differentiated (CAD) cell in late stage of differentiation stained for F-actin – white (cat no: A22287. ), microtubules – green (cat no: ab4074.). The cells were fixed and permeabilized with 3% paraformaldehyde, 0.1% glutaraldehyde, 4% sucrose, 0.5% Triton X-100. Objective Lens Magnification: 100x., Super-resolution spinning disk confocal image of a cath.a-differentiated (CAD) cell stained for F-actin – yellow (cat no: A22287.), microtubules – cyan (cat no: ab7291. ), and mitochondria – magenta (cat no: 11802-1-AP. Proteintech). The cell was fixed and permeabilized with 3% paraformaldehyde, 0.1% glutaraldehyde, 4% sucrose, 0.5% Triton X-100. Objective Lens Magnification: 100x.

Rebecca Lee: Yale University

Intestinal villi of the mouse small intestine coming out toward the viewer (outlined in magenta) with epithelial cells outlined in yellow/green. ​

1 and 3. antibodies used: EpCAM (#118202; yellow/green), CD29 ( #102201; magenta), DAPI ( #D9542; blue), 647 Donkey anti Rabbit ( #711-605-152), RRX Goat anti Hamster ( #127-295-160), 488 Donkey anti Rat ( #712-545-153)​

2. Mouse small intestine (jejunum), 4% PFA fixed​

4. Image taken on a confocal microscope ​

5. 25x water objective used

Category 1 winner (Featuring Proteintech products):

Rachel Stubler

Medical University of South Carolina

Category 2 winner (Featuring any products):

Wen Lu

Northwestern University Feinberg School of Medicine

Multiplexed immunofluorescence image of 5dpf transgenic zebrafish retina using IBEX imaging. 

Rat lumbar spinal cord neurons and motor neurons.

Mouse retina.

Embryonic mouse lung and trachea.