Combining deep nanoscopy and optical clearing, you can now image the ultrastructure of the glomerular filtration barrier from kidney tissue. The Leica TCS SP8 STED 3X (STimulated Emission Depletion) nanoscope sheds light on details deep inside the specimen with unprecedented three-dimensional detail. Using the STED WHITE glycerol objective, 3D nanoscopy in cleared tissue deeper than 150 µm sheds new insights in kidney disease.
Top image: Cleared kidney sample, revealing the podocyte cell layer; magnified podocytes by STED and electron microscopy (EM). Cleared kidney sample courtesy: David Unnersjö-Jess, KTH, Stockholm, Sweden. Scanning EM image of kidney podocytes courtesy: Center for Microscopy and Image Analysis, Univ. of Zurich, Dr. Urs Ziegler. Scale bar is 1 µm.
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The podocyte cell layer in the glomerular filtration barrier plays a central role in kidney disease. Alterations in the architecture of the podocytes are linked to the onset of renal failure. Therefore, the ability to image this layer at high resolution is key to identify healthy and unhealthy tissue.
The size of the podocyte layer requires imaging techniques with a resolution well below the diffraction limit. This has been typically addressed with electron microscopy techniques, demanding complex sample preparation protocols and where three dimensional information is missing.
3D STED Deep Nanoscopy opens a new door for kidney research. The combination of 3D STED Nanoscopy with the Leica TCS SP8 STED 3X and optical clearing enables imaging the kidney with unprecedented three-dimensional detail in light microscopy, and with protein specificity.