Name: quantifying glomerular permeability of fluorescent macromolecules using 2-photon microscopy in munich wistar rats
Uploaded: 2013-04-17T12:00:00
Description: Watch this Scientific Journal Video about Quantifying Glomerular Permeability of Fluorescent Macromolecules Using 2-Photon Microscopy in Munich Wistar Rats at JoVE.com

लेखकों शिरापरक पहुँच लाइनों की नियुक्ति से जुड़े सर्जिकल प्रक्रियाओं को पूरा करने के लिए डीआरएस सिल्विया बी कैम्पोस-Bilderback और जॉर्ज जे रोड्स को धन्यवाद देना चाहूंगा. उन्होंने यह भी Simonsen और Frömter उपभेदों दोनों से मिलकर म्यूनिख Wistar कालोनियों को बनाए रखने के लिए सारा ई छुड़ाना धन्यवाद देना चाहूंगा.

1. Sulfo-Rhodamine 101 Sulfonyl क्लोराइड (टेक्सास रेड) को Albumin चूहा सीरम का संयुग्मन <ol><li> अंतिम एकाग्रता एक 50 मिलीलीटर शंक्वाकार ट्यूब में 15 मिलीग्राम / एमएल, 100 मिमी सोडियम बिकारबोनिट पीएच 9.0 का चूहा में (आरएसए) सीरम albumin 6.667 मिली?...

<ol>
	<li>Russo, L. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+normal+kidney+filters+nephritic+levels+of+albumin+retrieved+by+proximal+tubule+cells;+retrieval+is+disrupted+in+nephritic+states.">The normal kidney filters nephritic levels of albumin retrieved by proximal tubule cells; retrieval is disrupted in nephritic states.</a> Kidney International. 71, 504 (2007).</li><li>Russo, L. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+tubular+uptake+explains+albuminuria+in+early+diabetic+nephropathy.">Impaired tubular uptake explains albuminuria in early diabetic nephropathy.</a> Journal of the American Society of Nephrology. 20 (3), 489 (2009).</li><li>Sandoval, R. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+factors+influence+glomerular+albumin+permeability+in+rats.">Multiple factors influence glomerular albumin permeability in rats.</a> Journal of the American Society of Nephrology. 23 (3), 447 (2012).</li><li>Tojo, A., Endou, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrarenal+handling+of+proteins+in+rats+using+fractional+micropuncture+technique.">Intrarenal handling of proteins in rats using fractional micropuncture technique.</a> American Journal of Physiology. 263, 601 (1992).</li><li>Asgeirsson, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glomerular+sieving+of+three+neutral+polysaccharides,+polyethylene+oxide+and+bikunin+in+rat:+Effects+of+molecular+size+and+conformation.">Glomerular sieving of three neutral polysaccharides, polyethylene oxide and bikunin in rat: Effects of molecular size and conformation.</a> Acta Physiologica. 191 (3), 237 (2007).</li><li>Sandoval, R. M., Molitoris, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+endocytosis+in+vivo+using+intravital+two-photon+microscopy.">Quantifying endocytosis in vivo using intravital two-photon microscopy.</a> Methods in Molecular Biology. 440, 389 (2008).</li><li>Dunn, K. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live-animal+imaging+of+renal+function+by+multi-photon+microscopy.">Live-animal imaging of renal function by multi-photon microscopy.</a> Curr. Protoc. Cytom. Chapter 12, Unit 12.9 (2007).</li></ol>

यहाँ पर प्रकाश डाला कदम हम वे निम्नलिखित नुकसान को दरकिनार क्योंकि सुसंगत और सटीक पारगम्यता मूल्यों का उत्पादन होगा कि लोगों को हो क्या महसूस प्रतिनिधित्व करते हैं: <ol><li> छितराया: अब तरंगदैर्य फोटॉनो?...

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 <td>Olympus Floview 1000 confocal/Multi-photon microscope</td>
 <td>Olympus America</td>
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 <td>Filters for detectors: Blue 430/100, Green 525/50, Red 605/90</td>
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 <td>Mode-Locked Ti:Sapphire Mai Tai Laser</td>
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quantifying glomerular permeability of fluorescent macromolecules using 2-photon microscopy in munich wistar rats

Kidney diseases involving urinary loss of large essential macromolecules, such as serum albumin, have long been thought to be caused by alterations in the permeability barrier comprised of podocytes, vascular endothelial cells, and a basement membrane working in unison. Data from our laboratory using intravital 2-photon microscopy revealed a more permeable glomerular filtration barrier (GFB) than previously thought under physiologic conditions, with retrieval of filtered albumin occurring in an early subset of cells called proximal tubule cells (PTC)1,2,3.
Previous techniques used to study renal filtration and establishing the characteristic of the filtration barrier involved micropuncture of the lumen of these early tubular segments with sampling of the fluid content and analysis4. These studies determined albumin concentration in the luminal fluid to be virtually non-existent; corresponding closely to what is normally detected in the urine. However, characterization of dextran polymers with defined sizes by this technique revealed those of a size similar to serum albumin had higher levels in the tubular lumen and urine; suggesting increased permeability5.
Herein is a detailed outline of the technique used to directly visualize and quantify glomerular fluorescent albumin permeability in vivo. This method allows for detection of filtered albumin across the filtration barrier into Bowman's space (the initial chamber of urinary filtration); and also allows quantification of albumin reabsorption by proximal tubules and visualization of subsequent albumin transcytosis6. The absence of fluorescent albumin along later tubular segments en route to the bladder highlights the efficiency of the retrieval pathway in the earlier proximal tubule segments. Moreover, when this technique was applied to determine permeability of dextrans having a similar size to albumin virtually identical permeability values were reported2. These observations directly support the need to expand the focus of many proteinuric renal diseases to included alterations in proximal tubule cell reclamation.

चित्रा 3 एक म्यूनिख Wistar Frömter चूहा और फ्लोरोसेंट एल्बुमिन की पारगम्यता निर्धारित करने के लिए उठाए गए कदमों की एक सतह ग्लोमेर्युल्स से ली गई एक छवि का एक उदाहरण दिखाता है. 0.0111 के एल्बुमिन के लिए GSC मूल्य...

Watch this Scientific Journal Video about Quantifying Glomerular Permeability of Fluorescent Macromolecules Using 2-Photon Microscopy in Munich Wistar Rats at JoVE.com

Quantifying Glomerular Permeability of Fluorescent Macromolecules Using 2-Photon Microscopy in Munich Wistar Rats

2 फोटॉन माइक्रोस्कोपी उच्च संकल्प intavital उपयोग एक तकनीक सीधे कल्पना और सतह ग्लोमेरुली में gloemrular निस्पंदन यों को. इस विधि में सामान्य और रोगग्रस्त दोनों राज्यों में बड़े अणुओं की पारगम्यता विशेषताओं के प्रत्यक्ष निर्धारण के लिए अनुमति देता है.

म्यूनिख Wistar चूहों में 2 फोटॉन माइक्रोस्कोपी का उपयोग फ्लोरोसेंट अणुओं की ग्लोमेरुलर पारगम्यता बढ़ाता

Kidney  diseases involving urinary loss of large essential macromolecules, such as  serum albumin, have long been thought to be caused by alterations in ...

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