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

JoVE Journal
Biology

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JoVE Journal Biology

An Efficient and Fast Method for Labeling and Analyzing Mouse Glomeruli

マウス糸球体の標識と分析のための効率的で迅速な方法

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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.

この研究は、CUBIC透明マウス腎臓からの糸球体を標識および分析するための、使いやすく完全でシンプルな一連の方法を提示します。糸球体数や体積などのデータは、フルオレセインイソチオシアネート(FITC)-デキストラン、ライトシート蛍光顕微鏡(LSFM)、または一般的な共焦点顕微鏡とImarisなどのソフトウェアを使用して簡単かつ確実に取得できます。

従来の方法を使用した糸球体のラベリングとカウントは、複雑で退屈で時間がかかり、かさばる機器が必要になる場合があります。私たちの方法は、腎臓全体の糸球体を分析するための即時かつ直感的に正確な標識法を提供します。私たちの技術は、腎臓全体の糸球体を分析するための迅速、簡単、正確、そして安価な方法です。

特別な機器を必要としないため、どの研究室でも実施できます。これは、糸球体を標識するための画期的で迅速でシンプルな方法であり、腎臓全体のすべての糸球体を直感的に観察することができます。糸球体の量、体積、分布などの情報を提供することで、研究者が腎臓の研究に大きなブレークスルーをもたらすのを支援する大きな可能性を秘めています。

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