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Protocol for analyzing the microstructure of nuclear fiber layers by using structured illumination microscopy coupled with Arivis 4D

Year: 2025
Type: article
Abstract: Here, we present a protocol for quantitatively assessing the microstructure of nuclear fiber layers by immunolabeling of a nuclear skeleton protein lamin A through structured illumination microscopy (SIM) coupled with an innovative algorithm in Arivis 4D. We describe steps of sample preprocessing for SIM and imaging of the inner-nucleus skeleton network of lamin A. We then detail procedures for creating a novel algorithm in Arivis 4D software to quantitatively assess the microparameters of the nuclear skeleton in a high-resolution image. • Steps for immunolabeling of a nuclear skeleton protein lamin A • Instructions for structured illumination microscopy (SIM) and sample preprocessing • Algorithm creation for 3D image reconstruction of nuclear skeleton network Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for quantitatively assessing the microstructure of nuclear fiber layers by immunolabeling of a nuclear skeleton protein lamin A through structured illumination microscopy (SIM) coupled with an innovative algorithm in Arivis 4D. We describe steps of sample preprocessing for SIM and imaging of the inner-nucleus skeleton network of lamin A. We then detail procedures for creating a novel algorithm in Arivis 4D software to quantitatively assess the microparameters of the nuclear skeleton in a high-resolution image.
DOI: 10.1016/j.xpro.2025.104308
Language: English
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W7117760882
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Cites: 10
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