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The 2024 Thermo Fisher Scientific Microscopy Imaging Contest has concluded, and we have selected the winners. We received submissions from a diverse range of scientific disciplines, including developmental research, cancer research, and neuroscience. Each entry showcased the inherent beauty of their respective fields through captivating images and provided a unique perspective into their research project. Thank you to all our partcipants.
Thermo Fisher Scientific Microscopy Imaging Contest 2024 Winners
What does the image represent?
Congenital diaphragmatic hernia (CDH) is a rare malformation that mainly affects lung development, and its disease mechanisms are poorly understood. This image shows a lung organoid, which is a miniature replica of a human lung, grown from stem cells derived from patient with CDH. In my work I utilise fetal fluids, such as amniotic fluid, to isolate tissue-specific stem cells and grow them into organoids. Lung organoids provide a close-up view of how lungs work at a cellular level, and can be used to study congenital lung diseases and find potential therapies.
What are the experimental details?
Antibodies, reagents, and imaging platform used: Hoechst 33342, Thermo Fisher Scientific (Cat. No.: 62249 ); Alexa Fluor Phalloidin 488, Thermo Fisher Scientific (Cat. No.: A12379 ); Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 647, Thermo Fisher Scientific (Cat. No.: A31573 ). Zeiss Confocal LSM710. Imaging software: Zeiss Zen.
What does the image represent?
To model and study the role glycans (sugars) play in driving Inflammatory Bowel Disease, we used a mouse model that expresses CreERT2 in the intestine. CreERT2 recognizes two LoxP sites flanking a genetic region of interest (C1galt1) and removes the loci from the mouse's genome upon induction with Tamoxifen (intraperitoneal injection). These mice are interbred with C3gnt-/- mice (global deletion) to create Double Knockout (DKO) mice that lack both Core 1 and Core 3 O-glycans when induced, which inhibits the mice from synthesizing mucins with complex sugars. The image is a section of the colon from this mouse model. Mice with a complete loss of both types of complex O-glycans can be confirmed via lack of Maackia amurensis lactin II (MALII) signal (red).
What are the experimental details?
Harvested colons were placed in Carnoy's fixative overnight at 4C and then processed. The tissue was stained with Maackia amurensis lectin II (MALII)-biotin diluted 1:500 for 1 hour at room temp. Slides were washed with PBST 3 times, then incubated with SYBR green (1:10,000 dilution), Wheat germ agglutinin (WGA)-350 blue (1:500 dilution), and Strep-594 red (1:500 dilution) for 1 hour at room temp. Slides were then washed again with PBST, then mounted with ProLong Glass Antifade Mountant. Blocking was done with the Endogenous Biotin-Blocking Kit following manufacturers recommendations.
Antibodies, reagents, and imaging platform used: Endogenous Biotin-Blocking Kit, Thermo Fisher (Cat. No.: E21390 ); ProLong Glass Antifade Mountant, Thermo Fisher (Cat. No.: P36982 ); Wheat Germ Agglutinin (WGA)-Alexa Fluor 350, Thermo Fisher (Cat. No.: W11263 ); Streptavidin, Alexa Fluor 594 conjugate, Thermo Fisher (Cat. No.: S11227 ); Maackia amurensis Lectin II (MAL II), Biotinylated, Vector Laboratories (Cat. No.: B-1265-1); SYBR Safe DNA Gel Stain, Thermo Fisher (Cat. No.: S33102 ); Thermo Fisher EVOS M5000.
What does the image represent?
The image shows a mouse neuroblastoma cath.a-differentiated (CAD) cell stained for F-actin (blue), microtubules (red), and mitochondria (green). It is part of a recent investigation where we demonstrated that prolonged depletion of F-actin increases the number of microtubules and causes them to become hyperacetylated, which alters the transport of mitochondria and other organelles.
My research focuses on studying the cytoskeleton crosstalk in neurodegenerative diseases. Cytoskeletal dynamics are essential for processes like axonal transport, synaptic plasticity, and cellular signaling, all of which are vital for healthy neuronal activity. In neurodegenerative diseases, disruptions in these processes can lead to neuronal dysfunction and cell death.
What are the experimental details?
The image was acquired with a super-resolution spinning-disk confocal microscope using optical photon reassignment. A Z-stack was acquired using a 100X objective and then deconvolved to further enhance the resolution.
Antibodies, reagents, and imaging platform used: Alexa Fluor 647 Phalloidin, Thermo Fisher Scientific (Cat. No. A22287); ProLong Diamond Anti-Fade, Thermo Fisher Scientific (Cat. No. P36961); Goat anti-mouse 488, Thermo Fisher Scientific (Cat. No. A11029); Goat anti-rabbit 568, Thermo Fisher Scientific (Cat. No. A11011); Magnesium chloride, Fisher Scientific (Cat. No. BP214); DMEM/F12 medium, Thermo Fisher Scientific (Cat. No. 11330-032); Cath.a-differentiated (CAD) cell (ATCC); rabbit polyclonal anti-alpha tubulin, Abcam (Cat. No. ab4074) and mouse monoclonal anti-TOM20 (4F3), Abcam (Cat. No. ab56783); Nikon CSU-W1 SoRa Spinning Disk Confocal Microscope.
What does the image represent?
In the last year of my PhD program, I received a diagnosis for a rare genetic kidney disease. This diagnosis, along with the required treatments, sparked a newfound interest in understanding the molecular mechanisms underlying kidney and glomerular diseases. As a result, I made a significant shift in my research focus from studying the cell biology of developmental cardiology/biology to investigating the genetic and molecular factors that contribute to chronic and acquired kidney and glomerular diseases.
The primary focus of my research lies in the investigation of a cellular structure called the primary cilium. This tiny, sensory organelle plays a crucial role in facilitating cell-to-cell communication and regulating various cellular processes such as differentiation, proliferation/apoptosis, cell survival, migration, and polarity. Although primary cilia were initially believed to be vestigial in nature, they were first discovered in the kidney over a century ago.
In the past five decades, significant advancements have been made in understanding the importance of primary cilia, as genetic mutations and structural damage to these organelles have been identified as causal factors in a group of diseases known as ciliopathies. This category of diseases continues to expand and encompasses a wide range of conditions, including obesity, osteoarthritis, certain forms of cancer, as well as congenital abnormalities and diseases such as Joubert Syndrome (JS), Bardet-Biedl syndrome (BBS), and the polycystic kidney diseases (PKD, ADPKD, ARPKD). The focus of my postdoctoral research training focuses on improving our understanding of the molecular mechanisms controlling glomerular (kidney) function, including those mediated through the primary cilium as well as mechanisms mediated by the sensory innervation of the glomerulus.
This tissue sample was stained to exhibit proteins fibronectin-1 and Sonic Hedgehog in glomeruli affected by minimal change disease glomerulopathy/podocytopathy.
What are the experimental details?
Formalin-fixed, paraffin-embedded human renal biopsy (FFPE) tissues were obtained from the Maine Health Biobank were processed and stained for structural markers found in healthy glomeruli. Primary antibodies were diluted to the appropriate working concentration and then applied to the tissue sections. A mountant with DAPI was applied to protect the fluorophore staining and mark the nuclei. The slides were imaged using Leica confocal microscopy at a resolution of 1024 x 1024 Hz. Z-stacks of the tissue sections were generated by capturing individual images at 0.2 micrometer z-thickness increments. A 63X oil objective with a zoom factor of 2.50 was used for imaging.
Antibodies, reagents, and imaging platform used: Anti-shh Polyclonal Antibody, Thermo Fisher Scientific (Cat. No.: MA5-51177 ); Alexa Fluor 488 goat anti-rabbit, Thermo Fisher Scientific (Cat. No.: A32731 ); Alexa Fluor 555 goat anti-mouse Thermo Fisher Scientific (Cat. No.: A21422 ). Fluormount with DAPI, anti-Fibronectin-1, Vendor: ProteinTech, Cat. No.: 15613-1-AP. Leica confocal microscope.
What is the most useful reagent for your experiment?
“The most useful reagents to these pivotal first imaging experiments were the secondary antibodies/Alexa fluorophores purchased through Invitrogen. Kidney tissue is known to be more difficult to work with using immunofluorescence due to a higher level of endogenously autofluorescence, a challenge that can be further exacerbated with the added influence of/variability of FFPE fixation/tissue preparation methods. Procuring secondary antibodies that are both robust and provide specific signal with minimal background ultimately enabled a clear visualization of human kidney/glomerular tissue in a manner that helped us characterize our experimental variables of interest as a part of designing a larger, ongoing study.”
- Lindsey Avery Fitzsimons
What does the image represent?
1 month old human neural network fibres (DcX positive, green) radiating from central neurosphere (Hoechst, blue) with GAP43 (red) positive cells.
This work is a part of Dr. Hodgett's laboratory at the Spinal Cord Repair Laboratory at the School of Human Sciences at UWA and Perron Institute for Neurological and Translational Science. Professor Hodgetts has over 20 years of expertise in cell based transplantation therapies. He has been devoted to this research since joining UWA in 1998, originally conducting research in neuromuscular diseases such as muscular dystrophy.
Since 2004 he has focused on the repair of the injured spinal cord using a variety of different stem cell, gene therapy, in vivo reprogramming, tissue engineering, neuroprotective and non-invasive strategies (such as repetitive transcranial magnetic stimulation and the use of infra-red/near infra-red light). His Spinal Cord Repair Laboratory acts as a core facility for other neuroscience researchers designed to promote neuro-regeneration into the setting of SCI toward preclinical studies. He is an academic coordinator for several undergraduate units at UWA, including Neuroscience, with supervision of over 35 Honours, 9 Masters and 20 PhD students.
He was the WA Representative for Australasian Neuroscience Society (2014-2017) and is co-chair for the ANS 2024 meeting in Perth. With previous NHMRC and current ARC funding, he has secured funds totaling nearly $5.5 million as Chief Investigator since 2001. He also works closely with national and international bio-artists (e.g., “cellF”, “Revivification”), as well as being scientific consultant for the internationally renowned “SymbioticA”.
What are the experimental details?
My research experiments use a variety of stem cell, gene therapy, bio-engineering and neuroprotection strategies to repair the injured spinal cord. These cells were stained and then imaged on a Nikon Eclipse Ti (Inverted) at x40 magnification. No image processing.
Antibodies, reagents, and imaging platform used: Doublecortin Monoclonal Antibody (2G5), Thermo Fisher Scientific (Cat. No.: MA5-17066); GAP43, Vendor: Thermo Fisher Scientific (Cat. No.: PA5-110781 replaced with PA5-34943 ); Nikon Eclipse Ti.
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