Physiology Images taken with THUNDER Imagers
See how Leica Microsystems instruments can help imaging cardiac organoids used to develop pacemaker cells for heart arrhythmias, muscular dystrophy, diabetes, sensory hair cell regeneration, cellular eye morphogenesis, retina and the vascular network, the Drosophila eye, HeLa cells, zebrafish heart, etc. There are different model organisms and specimens involved in this collection, such as chicken, mice, C. elegans, Drosophila, zebrafish, Arabidopsis thaliana, and HeLa cells.
Cardiac Spheroids
![Developing Heart Pacemaker Cells from Cardiac Spheroids Developing Heart Pacemaker Cells from Cardiac Spheroids. Cardiac Spheroid shown here as a maximum projection of the raw widefield image data (left) and THUNDER image after Instant Computational Clearing (right). The images are derived from approximately 60 µm Z stacks. Different stains (alpha actin and vimentin) are used to help identify the various cell types. Images courtesy of Sandra Grijalva, Wallace H. Coulter Department of Biomedical Engineering, Georgia Tech and Emory University, Atlanta, USA.](/fileadmin/_processed_/5/8/csm_Spheroid_shown_here_as_a_maximum_projection_a5a49cc032.jpg)
Vesicle formation, movement, and fusion
This video shows how observation of intracellular membrane trafficking along microtubules is enhanced with THUNDER Computational Clearing. Shown is a time-lapse movie of living HeLa cells transfected with Rab5a-GFP and stained with SIR-Tubulin. Vesicle formation, movement, and fusion can be observed.
Zebrafish Heart
![Zebrafish heart Zebrafish heart, DAPI (nuclei, blue), Tropomyosin (cardiomyocytes, red) and GFP (primordial cardiac layer, green). Courtesy of Anna Jazwinska, University of Fribourg, Switzerland.](/fileadmin/_processed_/e/c/csm_THUNDER_Imager_Zebrafish_heart_37c7f31c00.jpg)
HeLa Cells
Mouse Retina
Dr. Jiyeon Lee, a scientist at Roche Genentech in South San Francisco, CA uses the mouse retina model to study the interaction between endothelial cells, blood vessels, and astrocytesin the retina. Dr. Lee uses whole mount retina preparation to visualize its vasculature. Following fixation and staining, the whole retina must be imaged at high resolution to provide an overview of the whole vascular network, as well as single cell interactions.
![Mouse retina Mouse retina was fixed and stained by following reagents: anti-CD31 antibody (green): Endothelia cells, IsoB4 (red): Blood vessels, and microglia anti-GFAP antibody (blue): Astrocytes Sample courtesy by Jeremy Burton, PhD and Jiyeon Lee, PhD, Genentech Inc., South San Francisco, USA. Imaged by Olga Davydenko, PhD (Leica).](/fileadmin/_processed_/0/c/csm_THUNDER_Imager_3D_Cell_Culture_Mouse_Retina_eeba2a8a4e.jpg)
Arabidopsis Thaliana Root
Murine Esophageal Organoids
![Murine esophageal organoids Murine esophageal organoids imaged with THUNDER Imager 3D Cell Culture. Integrin alpha6 (AlexaFluor 488, green), Sox2 (AlexaFluor 568, red), Nucleus (Dapi, blue). Sample courtesy of Dr. Fabio Tadeu Arroso Martins, Tampere University, Finland. Sample imaged by Janne Ylärinne, PhD, Immuno Diagnostic Oy.](/fileadmin/_processed_/d/2/csm_THUNDER-Imager_Murine_esophageal_Esophagus_Organoids_a8a9c8ee6a.jpg)
Adult Drosophila Muscle
Mouse Lens Section
![Transversal mouse adult lens section This slide is a transversal Mouse adult lens section showing the fiber lens cells organization (green, stained with anti-β-catenin antibody (BDB610153), a membrane protein) and Afadin (red, stained with anti-Afadin (PA1-25076), an actin filament-binding protein, blue, Hoechst). Image courtesy of Dr. Nathalie Houssin, Plagemen Lab, Ohio State University, USA.](/fileadmin/_processed_/d/e/csm_THUNDER_Imager_3D_Cell_Culture_Mouse-Lens-Section_Houssin_48465dc3a0.jpg)
C. elegans Gonades
Chicken Cochlea Tilescan
Dr. Amanda Janesick, a postdoctoral fellow in the laboratory of Dr. Stefan Heller, studies the development and regeneration of the inner ear, specifically the sensory hair cells, which serve a mechanosensitive purpose in the inner ear. Dr. Janesick uses the chicken embryo as a model to study sensory hair cell regeneration, because of the ability of chickens to naturally recover from hearing loss.
Dr. Janesick uses thick vibratome sections of post-hatch 7 day chicken cochlear tissue to visualize the sensory hair cells in the inner ear. Typically, this is a challenging sample for widefield fluorescence microscopes, due to the haze inherent to thicker tissue samples, which obscures structures of interest (Figure 1, left panel). Using a THUNDER Imager 3D Tissue, the haze and background was cleared from this image using Instant Computational Clearing (ICC, Figure 1, right panel), making it suitable for observation of individual sensory hair cells (magenta) and supporting cells (yellow) of the inner ear. This 3-channel 10-position tilescan took under a minute to acquire and process with ICC. Because ICC is a 2D method, and does not require z-stacks, it could be applied to this single plane tilescan.
![Chicken cochlea tilescan Chicken cochlea tilescan imaged on a THUNDER 3D Tissue Imager. The raw epifluorescence image is on the left, and the result of Instant Computational Clearing is on the right. This 43 µm thick vibratome section of a post-hatch day 7 chicken cochlea was stained with a nuclear DAPI stain (cyan), antibodies for Myosin 7a labeling sensory hair cells (magenta), and Sox2 labeling supporting cells (yellow). 40x oil immersion objective (NA 1.3) was used to acquire this 10-position tilescan.](/fileadmin/_processed_/d/5/csm_THUNDER_Imager_3D_Tissue_Chicken-cochlea_Janesick_c638659a70.jpg)
Transgenic Drosophila Photoreceptors of the Eye
Pancreatic Islet
A better understanding of how type 1 diabetes (T1D) develops is the first step to potentially develop new therapies capable of preventing or permanently reversing the disease. Due to the inaccessibility to human pancreatic tissue, our knowledge of the disease in humans is limited. The Network for Pancreatic Organ donors with Diabetes (nPOD) was established with the idea of providing valuable tissues from healthy and diabetic donors toanswer basic questions about the pathogenesis of T1D. One of the interests of the von Herrath lab is toidentifycytokines in the pancreatic tissue samples of T1D cases obtained from nPOD. Since T1D is an autoimmune disease, where insulin producing pancreatic beta cells are attacked by the immune system, understanding the cytokine milieu in the pancreas of T1D will lead to a better understanding of the disease pathogenesis and contribute towards development of therapeutic targets for T1D.
![EDoF reconstruction of an isolated human islet EDoF reconstruction of an isolated human islet to experimentally examine the expression of IL-17, a proinflammatory cytokine, in individual human islet cells. The images have the following markers: Insulin (AF488; green), Glucagon (AF555; red) and IL-17 (AF647; magenta) and Hoechst (nuclei; blue). Image courtesy of the Matthias Von Herrath Lab at the La Jolla Institute of Immunology, La Jolla, CA., USA.](/fileadmin/_processed_/9/f/csm_IL6-organoids-THUNDER-Thunder-Imager-3D-Cell-Culture_868d7731f9.jpg)
Mouse Dystrophin Staining on Muscle Fibres
![Mouse muscle tissue Top down view of mouse muscle tissue (total area: 0.56 mm2), illustrating the difference between the raw data and the data after THUNDER LVCC processing.](/fileadmin/_processed_/3/1/csm_THUNDER_Imager_3D_Cell_Culture_Mouse-Muscle_58a9169dac.jpg)