A1 Journal article (refereed)
Iterative immunostaining combined with expansion microscopy and image processing reveals nanoscopic network organization of nuclear lamina (2023)

Mäntylä, E., Montonen, T., Azzari, L., Mattola, S., Hannula, M., Vihinen-Ranta, M., Hyttinen, J., Vippola, M., Foi, A., Nymark, S., & Ihalainen, T. O. (2023). Iterative immunostaining combined with expansion microscopy and image processing reveals nanoscopic network organization of nuclear lamina. Molecular Biology of the Cell, 34(9). https://doi.org/10.1091/mbc.e22-09-0448

JYU authors or editors

Mattola, Salla
Vihinen-Ranta, Maija

Publication details

All authors or editors: Mäntylä, Elina; Montonen, Toni; Azzari, Lucio; Mattola, Salla; Hannula, Markus; Vihinen-Ranta, Maija; Hyttinen, Jari; Vippola, Minnamari; Foi, Alessandro; Nymark, Soile; et al.

Journal or series: Molecular Biology of the Cell

ISSN: 1059-1524

eISSN: 1939-4586

Publication year: 2023

Publication date: 21/06/2023

Volume: 34

Issue number: 9

Publisher: American Society for Cell Biology (ASCB)

Publication country: United States

Publication language: English

DOI: https://doi.org/10.1091/mbc.e22-09-0448

Publication open access: Openly available

Publication channel open access: Partially open access channel

Publication is parallel published (JYX): https://jyx.jyu.fi/handle/123456789/88580

Additional information: Brief Report

Abstract

Investigation of nuclear lamina architecture relies on super-resolved microscopy. However, epitope accessibility, labeling density, and detection precision of individual molecules pose challenges within the molecularly crowded nucleus. We developed iterative indirect immunofluorescence (IT–IF) staining approach combined with expansion microscopy (ExM) and structured illumination microscopy to improve super-resolution microscopy of subnuclear nanostructures like lamins. We prove that ExM is applicable in analyzing highly compacted nuclear multiprotein complexes such as viral capsids and provide technical improvements to ExM method including 3D-printed gel casting equipment. We show that in comparison to conventional immunostaining, IT-IF results in a higher signal-to-background –ratio and a mean fluorescence intensity by improving the labeling density. Moreover, we present a signal processing pipeline for noise estimation, denoising, and deblurring to aid in quantitative image analyses and provide this platform for the microscopy imaging community. Finally, we show the potential of signal-resolved IT–IF in quantitative super-resolution ExM imaging of nuclear lamina and reveal nanoscopic details of the lamin network organization - a prerequisite for studying intranuclear structural co-regulation of cell function and fate.

Keywords: cell nucleus; lamins; structure (properties); optical microscopy

Fields of science: