A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
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-tekijät tai -toimittajat

Mattola, Salla
Vihinen-Ranta, Maija

Julkaisun tiedot

Julkaisun kaikki tekijät tai toimittajat: Mäntylä, Elina; Montonen, Toni; Azzari, Lucio; Mattola, Salla; Hannula, Markus; Vihinen-Ranta, Maija; Hyttinen, Jari; Vippola, Minnamari; Foi, Alessandro; Nymark, Soile; et al.

Lehti tai sarja: Molecular Biology of the Cell

ISSN: 1059-1524

eISSN: 1939-4586

Julkaisuvuosi: 2023

Ilmestymispäivä: 21.06.2023

Volyymi: 34

Lehden numero: 9

Kustantaja: American Society for Cell Biology (ASCB)

Julkaisumaa: Yhdysvallat (USA)

Julkaisun kieli: englanti

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

Julkaisun avoin saatavuus: Avoimesti saatavilla

Julkaisukanavan avoin saatavuus: Osittain avoin julkaisukanava

Julkaisu on rinnakkaistallennettu (JYX): https://jyx.jyu.fi/handle/123456789/88580

Lisätietoja: Brief Report

Tiivistelmä

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.

YSO-asiasanat: tuma; lamiinit; rakenne (ominaisuudet); valomikroskopia

Liittyvät organisaatiot

Hankkeet, joissa julkaisu on tehty

OKM-raportointi: Kyllä

Raportointivuosi: 2023

Alustava JUFO-taso: 2