Serial intravital 2-photon microscopy and analysis of the kidney using upright microscopes

Donato Sardella; Anders M. Kristensen; Luca Bordoni; Hanne Kidmose; Ali Shahrokhtash; Duncan S. Sutherland; Sebastian Frische; Ina Maria Schiessl

doi:10.3389/fphys.2023.1176409

Serial intravital 2-photon microscopy and analysis of the kidney using upright microscopes

Donato Sardella^*, Anders M. Kristensen, Luca Bordoni, Hanne Kidmose, Ali Shahrokhtash, Duncan S. Sutherland, Sebastian Frische, Ina Maria Schiessl^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

4 Citations (Scopus)

Abstract

Serial intravital 2-photon microscopy of the kidney and other abdominal organs is a powerful technique to assess tissue function and structure simultaneously and over time. Thus, serial intravital microscopy can capture dynamic tissue changes during health and disease and holds great potential to characterize (patho-) physiological processes with subcellular resolution. However, successful image acquisition and analysis require significant expertise and impose multiple potential challenges. Abdominal organs are rhythmically displaced by breathing movements which hamper high-resolution imaging. Traditionally, kidney intravital imaging is performed on inverted microscopes where breathing movements are partly compensated by the weight of the animal pressing down. Here, we present a custom and easy-to-implement setup for intravital imaging of the kidney and other abdominal organs on upright microscopes. Furthermore, we provide image processing protocols and a new plugin for the free image analysis software FIJI to process multichannel fluorescence microscopy data. The proposed image processing pipelines cover multiple image denoising algorithms, sample drift correction using 2D registration, and alignment of serial imaging data collected over several weeks using landmark-based 3D registration. The provided tools aim to lower the barrier of entry to intravital microscopy of the kidney and are readily applicable by biomedical practitioners.

Original language	English
Article number	1176409
Journal	Frontiers in Physiology
Volume	14
Number of pages	17
ISSN	1664-042X
DOIs	https://doi.org/10.3389/fphys.2023.1176409
Publication status	Published - Apr 2023

Keywords

3D printing
animal holder
bioimage analysis
deep learning
image denoising
image registration
intravital microscopy (IVM)
multiphoton microcopy

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10.3389/fphys.2023.1176409Licence: CC BY

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title = "Serial intravital 2-photon microscopy and analysis of the kidney using upright microscopes",

abstract = "Serial intravital 2-photon microscopy of the kidney and other abdominal organs is a powerful technique to assess tissue function and structure simultaneously and over time. Thus, serial intravital microscopy can capture dynamic tissue changes during health and disease and holds great potential to characterize (patho-) physiological processes with subcellular resolution. However, successful image acquisition and analysis require significant expertise and impose multiple potential challenges. Abdominal organs are rhythmically displaced by breathing movements which hamper high-resolution imaging. Traditionally, kidney intravital imaging is performed on inverted microscopes where breathing movements are partly compensated by the weight of the animal pressing down. Here, we present a custom and easy-to-implement setup for intravital imaging of the kidney and other abdominal organs on upright microscopes. Furthermore, we provide image processing protocols and a new plugin for the free image analysis software FIJI to process multichannel fluorescence microscopy data. The proposed image processing pipelines cover multiple image denoising algorithms, sample drift correction using 2D registration, and alignment of serial imaging data collected over several weeks using landmark-based 3D registration. The provided tools aim to lower the barrier of entry to intravital microscopy of the kidney and are readily applicable by biomedical practitioners.",

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author = "Donato Sardella and Kristensen, {Anders M.} and Luca Bordoni and Hanne Kidmose and Ali Shahrokhtash and Sutherland, {Duncan S.} and Sebastian Frische and Schiessl, {Ina Maria}",

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doi = "10.3389/fphys.2023.1176409",

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AU - Kristensen, Anders M.

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AU - Kidmose, Hanne

AU - Shahrokhtash, Ali

AU - Sutherland, Duncan S.

AU - Frische, Sebastian

AU - Schiessl, Ina Maria

PY - 2023/4

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N2 - Serial intravital 2-photon microscopy of the kidney and other abdominal organs is a powerful technique to assess tissue function and structure simultaneously and over time. Thus, serial intravital microscopy can capture dynamic tissue changes during health and disease and holds great potential to characterize (patho-) physiological processes with subcellular resolution. However, successful image acquisition and analysis require significant expertise and impose multiple potential challenges. Abdominal organs are rhythmically displaced by breathing movements which hamper high-resolution imaging. Traditionally, kidney intravital imaging is performed on inverted microscopes where breathing movements are partly compensated by the weight of the animal pressing down. Here, we present a custom and easy-to-implement setup for intravital imaging of the kidney and other abdominal organs on upright microscopes. Furthermore, we provide image processing protocols and a new plugin for the free image analysis software FIJI to process multichannel fluorescence microscopy data. The proposed image processing pipelines cover multiple image denoising algorithms, sample drift correction using 2D registration, and alignment of serial imaging data collected over several weeks using landmark-based 3D registration. The provided tools aim to lower the barrier of entry to intravital microscopy of the kidney and are readily applicable by biomedical practitioners.

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KW - multiphoton microcopy

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Serial intravital 2-photon microscopy and analysis of the kidney using upright microscopes

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