Multicolour three dimensional structured illumination microscopy of immunolabeled plant microtubules and associated proteins

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F19%3A73596875" target="_blank" >RIV/61989592:15310/19:73596875 - isvavai.cz</a>

Result on the web

<a href="https://plantmethods.biomedcentral.com/track/pdf/10.1186/s13007-019-0406-z" target="_blank" >https://plantmethods.biomedcentral.com/track/pdf/10.1186/s13007-019-0406-z</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1186/s13007-019-0406-z" target="_blank" >10.1186/s13007-019-0406-z</a>

Result language

angličtina

Original language name

Multicolour three dimensional structured illumination microscopy of immunolabeled plant microtubules and associated proteins

Original language description

BackgroundIn the present work, we provide an account of structured illumination microscopy (SIM) imaging of fixed and immunolabeled plant probes. We take advantage of SIM, to superresolve intracellular structures at a considerable z-range and circumvent its low temporal resolution capacity during the study of living samples. Further, we validate the protocol for the imaging of fixed transgenic material expressing fluorescent protein-based markers of different subcellular structures.ResultsFocus is given on 3D imaging of bulky subcellular structures, such as mitotic and cytokinetic microtubule arrays as well as on the performance of SIM using multichannel imaging and the quantitative correlations that can be deduced. As a proof of concept, we provide a superresolution output on the organization of cortical microtubules in wild-type and mutant Arabidopsis cells, including aberrant preprophase microtubule bands and phragmoplasts in a cytoskeletal mutant devoid of the p60 subunit of the microtubule severing protein KATANIN and refined details of cytoskeletal aberrations in the mitogen activated protein kinase (MAPK) mutant mpk4. We further demonstrate, in a qualitative and quantitative manner, colocalizations between MPK6 and unknown dually phosphorylated and activated MAPK species and we follow the localization of the microtubule associated protein 65-3 (MAP65-3) in telophase and cytokinetic microtubular arrays.Conclusions3D SIM is a powerful, versatile and adaptable microscopy method for elucidating spatial relationships between subcellular compartments. Improved methods of sample preparation aiming to the compensation of refractive index mismatches, allow the use of 3D SIM in the documentation of complex plant cell structures, such as microtubule arrays and the elucidation of their interactions with microtubule associated proteins.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10601 - Cell biology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2019

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Plant Methods

ISSN

1746-4811

e-ISSN

—

Volume of the periodical

15

Issue of the periodical within the volume

MAR

Country of publishing house

GB - UNITED KINGDOM

Number of pages

17

Pages from-to

"22-1"-"22-17"

UT code for WoS article

000460703400001

EID of the result in the Scopus database

2-s2.0-85062709463