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University of Washington
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Transformative Tools for Imaging Tissue Ultrastructure in Diabetic Kidney Disease
This proposal seeks to apply and adapt the next-generation super-resolution optical microscopy method expansion microscopy (ExM) to the study of human diabetic kidney disease, the most common cause of end-stage renal disease in the U.S. In ExM, fluorescent labels on fixed specimens are linked to a swellable polymer hydrogel that is physically expanded with remarkably low distortion. The process allows features closer than the diffraction limit of light (~250 nm) to become resolvable in the expanded specimen and achieves ~65 nm spatial resolution on thick or thin specimens with ordinary confocal microscopes. This greatly improved spatial resolution is sufficient to reveal key ultrastructural features of the kidney including glomerular foot process width and glomerular basement membrane width while also providing molecular details such as extracellular membrane composition or presence of immune deposits, etc. Importantly, ExM makes use of standard optical microscopes that are already familiar and widely available, together with inexpensive sample processing methods, so that the method has the potential to become widely used in research and clinical settings for the study or diagnosis of kidney diseases, including DKD. Our specific aims are: 1) to develop and optimize procedures to study fresh kidney specimens using ExM; 2) to adapt these procedures to study archived paraffin-embedded tissue by ExM; and 3) to perform correlative ExM and electron microscopy imaging as a robust validation of our methodology.
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Transformative Tools for Imaging Tissue Ultrastructure in Diabetic Kidney Disease (Vaughan, Joshua)
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Year: 2019; Items: 3
Microscopy with ultraviolet surface excitation for wide-area pathology of breast surgical margins.
Xie W, Chen Y, Wang Y, Wei L, Yin C, Glaser AK, Fauver ME, Seibel EJ, Dintzis SM, Vaughan JC, Reder NP, Liu JTC
Journal of biomedical optics
, 2019 (24), 1 - 11
Dual lineage tracing shows that glomerular parietal epithelial cells can transdifferentiate toward the adult podocyte fate.
Kaverina NV, Eng DG, Freedman BS, Kutz JN, Chozinski TJ, Vaughan JC, Miner JH, Pippin JW, Shankland SJ
, 2019 (96), 597 - 611
Multi-immersion open-top light-sheet microscope for high-throughput imaging of cleared tissues.
Glaser AK, Reder NP, Chen Y, Yin C, Wei L, Kang S, Barner LA, Xie W, McCarty EF, Mao C, Halpern AR, Stoltzfus CR, Daniels JS, Gerner MY, Nicovich PR, Vaughan JC, True LD, Liu JTC
, 2019 (10), 2781
Year: 2018; Items: 1
Volumetric, Nanoscale Optical Imaging of Mouse and Human Kidney via Expansion Microscopy.
Chozinski TJ, Mao C, Halpern AR, Pippin JW, Shankland SJ, Alpers CE, Najafian B, Vaughan JC
, 2018 (8), 10396
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The DiaComp Steering Committee is the governing body of the consortium. The principle function of this committee is to guide the scientific direction of the consortium. This is accomplished by creating various subcommittees necessary to advance the scientific goals and providing guidance to the broader complications research community. Policies for the consortium are developed through consultation with the
External Evaluation Committee
The DiaComp Nephropathy Committee has the principal function of furthering the mission of the consortium with regard to diabetic kidney disease.
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Please acknowledge all posters, manuscripts or scientific materials that were generated in part or whole using funds from the Diabetic Complications Consortium(DiaComp) using the following text:
Financial support for this work provided by the NIDDK Diabetic Complications Consortium (RRID:SCR_001415, www.diacomp.org), grants DK076169 and DK115255
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