An Efficient and Fast Method for Labeling and Analyzing Mouse Glomeruli

Lin Bai; Yaping Wu; Wenshu Dai; Qiuxiao Shi; Lei Wu; Jie Zhang; Lily Zheng

doi:10.3791/65973

Une méthode efficace et rapide pour l’étiquetage et l’analyse des glomérules de souris

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An Efficient and Fast Method for Labeling and Analyzing Mouse Glomeruli

Une méthode efficace et rapide pour l’étiquetage et l’analyse des glomérules de souris

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09:50 min

February 9, 2024

DOI: 10.3791/65973-v

Lin Bai^1,2, Yaping Wu², Wenshu Dai¹, Qiuxiao Shi², Lei Wu², Jie Zhang^1,2, Lily Zheng³

¹Key Laboratory of Transplant Engineering and Immunology, NHFPC, West China Hospital,Sichuan University, ²Core Facility of West China Hospital,Sichuan University, ³Regenerative Medicine Research Center, West China Hospital,Sichuan University

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Overview

This study presents a groundbreaking method for labeling and analyzing glomeruli in CUBIC-cleared mouse kidneys, aiming to simplify the analysis of kidney structures. Utilizing fluorescein isothiocyanate (FITC)-Dextran and various microscopy techniques, researchers can effortlessly quantify glomerulus number and volume across the entire kidney.

Key Study Components

Research Area

Kidney structure analysis
Glomeruli labeling techniques
Light sheet and confocal microscopy

Background

Traditional glomeruli analysis methods are often complex and time-consuming.
Simplifying the process can facilitate significant research breakthroughs.
Current methodologies may require specialized equipment that is not universally available.

Methods Used

Fluorescein isothiocyanate (FITC)-Dextran labeling
Mouse kidneys as the biological model
Light sheet fluorescence microscopy (LSFM) and confocal microscopy

Main Results

A reliable and quick method for glomeruli analysis was established.
Quantitative data on glomeruli number, volume, and distribution can be obtained.
The methodology offers potential applications in kidney disease research.

Conclusions

The study demonstrates a rapid and intuitive approach for glomeruli analysis.
It holds significant relevance for advancing kidney biology research.

Frequently Asked Questions

What is the significance of analyzing glomeruli?

Analyzing glomeruli is crucial for understanding kidney function and disease mechanisms.

Can this method be used in other types of tissues?

While this method is tailored for kidney tissues, adaptations may allow its use in other transparent tissues.

What microscopic techniques are utilized in this study?

Light sheet fluorescence microscopy and confocal microscopy are employed for detailed imaging.

Is specialized equipment required for this method?

No, this method is designed to be accessible and does not necessitate specialized equipment.

How does this method improve upon traditional glomeruli analysis?

It simplifies the labeling and counting process, making it faster and more intuitive.

What types of data can be obtained from this analysis?

Researchers can obtain data on glomeruli quantity, volume, and distribution.

What future research directions does this study suggest?

Further exploration into labeling biotargets in transparent tissues is planned to meet evolving research needs.

Cette étude présente un ensemble de méthodes simples et faciles à utiliser pour marquer et analyser les glomérules des reins de souris nettoyés par CUBIC. Des données telles que le nombre et le volume du glomérule peuvent être obtenues facilement et de manière fiable à l’aide de l’isothiocyanate de fluorescéine (FITC)-Dextran, de la microscopie à fluorescence à feuille de lumière (LSFM) ou de la microscopie confocale commune et de logiciels tels qu’Imaris.

L’étiquetage et le comptage des glomérules à l’aide de méthodes conventionnelles peuvent être compliqués, fastidieux, longs et des instruments encombrants sont nécessaires. Notre méthode offre une méthode de marquage instantanée et intuitive pour l’analyse des glomérules dans l’ensemble du rein. Notre technique est une méthode rapide, facile, précise et peu coûteuse pour analyser les glomérules dans l’ensemble du rein.

Il ne nécessite aucun équipement spécial, il peut donc être réalisé dans n’importe quel laboratoire. Il s’agit d’une méthode révolutionnaire, rapide et simple pour marquer les glomérules, permettant une observation intuitive de tous les glomérules dans l’ensemble du rein. Il a un grand potentiel pour aider les chercheurs à faire des percées significatives dans l’étude du rein en fournissant des informations telles que la quantité, le volume, la distribution des glomérules, etc.

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