Glomeruli Isolation
Glomeruli Isolation - Related Videos
03:35
Glomeruli Isolation: A Technique to Obtain Intact Glomeruli from Murine Kidney
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2023
This video describes the method to isolate intact glomeruli from mouse kidneys. The encapsulated and decapsulated glomeruli thus obtained can further be used for culturing parietal epithelial cell outgrowths from the murine kidney.
12:19
Single-channel Analysis and Calcium Imaging in the Podocytes of the Freshly Isolated Glomeruli
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Cited by 25 •
2015
Changes in the intracellular calcium levels in the podocytes are one of the most important means to control the filtration function of glomeruli. Here we explain a high-throughput approach that allows detection of real-time calcium handling and single ion channels activity in the podocytes of the freshly isolated glomeruli.
10:14
An Efficient Sieving Method to Isolate Intact Glomeruli from Adult Rat Kidney
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Cited by 13 •
2018
The main focus of this protocol is to efficiently isolate viable primary glomeruli cultures with minimal contaminants for use in a variety of downstream applications. The isolated glomeruli retain structural relationships between component cell types and can be cultured ex vivo for a short time.
09:12
Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins
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Cited by 4 •
2019
Here we present a protocol for murine in vivo labeling of glomerular cell surface proteins with biotin. This protocol contains information on how to perfuse mouse kidneys, isolate glomeruli, and perform endogenous immunoprecipitation of the protein of interest.
09:50
An Efficient and Fast Method for Labeling and Analyzing Mouse Glomeruli
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2024
This study presents an easy-to-use, complete, and simple set of methods to label and analyze glomeruli from CUBIC-cleared mouse kidneys. Data such as glomerulus number and volume can be obtained easily and reliably using fluorescein isothiocyanate (FITC)-Dextran, light sheet fluorescence microscopy (LSFM), or common confocal microscopy and software such as Imaris.
FAQs
Mechanical disruption breaks the kidney tissue into separable components without relying on a single purification step. Filtration or sieving then sorts material according to particle size, while differential centrifugation further enriches fractions based on how components separate during spinning. Together, these operations reduce tubular fragments and debris while retaining a fraction enriched in glomeruli for downstream study.
Glomeruli and surrounding renal structures differ in both organization and size. Those differences allow sieving or filtration to distinguish larger, more intact glomerular structures from smaller tubular fragments and debris. Differential centrifugation adds another separation principle, helping improve enrichment when mechanical disruption produces a mixed tissue suspension.
An isolated preparation can support focused examination of podocytes, endothelial cells, and basement membranes, the components associated with the glomerular filtration barrier. Retaining these structures makes the sample useful for microscopic, biochemical, and molecular analyses. Their examination can connect cellular or molecular changes with filtration-barrier organization and function.
A typical workflow begins by mechanically disrupting renal tissue, followed by filtration or sieving to separate structures by size. Differential centrifugation is then used to enrich the glomerular fraction and remove smaller tubular material and debris. The resulting preparation can be directed to microscopy, biochemical measurements, or molecular analysis, depending on the research question.
The separation strategy must produce enough tissue disruption to release glomeruli from surrounding renal tissue while still supporting enrichment of intact structures. Filtration, sieving, and centrifugation each contribute differently to removing unwanted material. The resulting balance influences whether the preparation is suitable for examining glomerular architecture, barrier components, or molecular features.
Researchers use isolated glomeruli when they need focused access to the filtration units rather than mixed kidney tissue. The preparation supports studies of kidney development, filtration function, disease mechanisms, and responses to potential treatments. It can also provide material for microscopic, biochemical, and molecular analyses centered on the glomerular filtration barrier.