Super-Resolution Microscopy: A Technique to Revolutionize Research and Diagnosis of Glomerulopathies

Abstract

For decades, knowledge about glomerular (patho)physiology has been tightly linked with advances in microscopic imaging technology. For example, the invention of electron microscopy was required to hypothesize about the mode of glomerular filtration barrier function. Super-resolution techniques, defined as fluorescence microscopy approaches that surpass the optical resolution limit of around 200 nm, have been made available to the scientific community. Several of these different techniques currently are in use in glomerular research. Using three-dimensional structured illumination microscopy, the exact morphology of the podocyte filtration slit can be morphometrically analyzed and quantitatively compared across samples originating from animal models or human biopsies. Several quantitative image analysis approaches and their potential influence on glomerular research and diagnostics are discussed.

Ultrastructure expansion microscopy (U-ExM) of mouse and human kidneys for analysis of subcellular structures

“…Several approaches to expand and immunolabel kidneys have been developed using the ExM, MAP, ExPath and Magnify methods (Bucur & Zhao, 2018; Chozinski et al, 2018; Klimas et al, 2023; Ku et al, 2016; Kylies et al, 2023; Siegerist et al, 2022; Unnersjö-Jess et al, 2021; Unnersjö-Jess et al, 2018; Wunderlich et al, 2021; Zhao et al, 2017). However, there are several advantages to the U-ExM approach: a) First, U-ExM allows for post-expansion staining of the sample.…”

“…Importantly, it eliminates the need for access to costly super-resolution microscopes to achieve sub diffraction-resolution fluorescence imaging of cellular structures. Although expansion microscopy was only recently established as a new imaging modality, several protocols have now been developed for expanding cells and tissues from a variety of organisms including bacteria, parasites, insects and vertebrates (Atchou et al, 2023; Bai et al, 2023; Bandeira et al, 2023; Campbell et al, 2021; Chang et al, 2024; Chang et al, 2017; Chen et al, 2015; Cheng et al, 2023; Ching et al, 2022; Chozinski et al, 2016; Chozinski et al, 2018; Damstra et al, 2022; Damstra et al, 2023; Dos Santos Pacheco & Soldati-Favre, 2021; Fan et al, 2021; Gallagher & Zhao, 2021; Gambarotto et al, 2021; Gambarotto et al, 2019; Gaudreau-Lapierre et al, 2021; Hamel & Guichard, 2021; Jurriens et al, 2021; Klimas et al, 2023; Kraft et al, 2023; Ku et al, 2016; Kunz et al, 2019; Kunz et al, 2021; Liffner et al, 2023; Lin et al, 2022; Louvel et al, 2023; Mäntylä et al, 2023; Middelhauve et al, 2023; Moye et al, 2023; Murakami et al, 2018; Park et al, 2019; Park et al, 2021; Parveen et al, 2023; Perelsman et al, 2022; Pernal et al, 2020; Pesce et al, 2019; Ponjavić et al, 2021; Rodriguez-Gatica et al, 2022; Sahabandu et al, 2019; Siegerist et al, 2022; Steib et al, 2022; Tillberg & Chen, 2019; Tillberg et al, 2016; Unnersjö-Jess et al, 2021; Unnersjö-Jess et al, 2018; Wainman, 2021; Wang et al, 2018; Wassie et al, 2019; Wen et al, 2023; Wilmerding et al, 2023; Woo et al, 2020; Yu et al, 2022; Zhao et al, 2017; Zhu et al, 2021; Zhuang & Shi, 2024).…”

Super-resolution histology of paraffin-embedded samples via photonic chip-based microscopy

“…STED, despite being a reconstruction-free method, is a point-scanning technique with low throughput for scanning the centimeter-scale tissue areas commonly used in histology. Furthermore, the light scattering aberrations experienced by the depletion laser ultimately compromises the lateral resolution achievable with STED in FFPE samples 24 . The FF-SRM comprises a set of statistical methods capable of resolving fine structures out of conventional wide-field image stacks, without the need for special equipment or complex sample preparation.…”