Počet záznamů: 1
Correlative light-environmental scanning electron microscopy of plasma membrane efflux carriers of plant hormone auxin
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SYSNO ASEP 0546239 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Correlative light-environmental scanning electron microscopy of plasma membrane efflux carriers of plant hormone auxin Tvůrce(i) Stelate, A. (CZ)
Tihlaříková, Eva (UPT-D) RID, ORCID, SAI
Schwarzerová, K. (CZ)
Neděla, Vilém (UPT-D) RID, ORCID, SAI
Petrášek, J. (CZ)Celkový počet autorů 5 Číslo článku 1407 Zdroj.dok. Biomolecules. - : MDPI
Roč. 11, č. 10 (2021)Poč.str. 13 s. Forma vydání Online - E Jazyk dok. eng - angličtina Země vyd. CH - Švýcarsko Klíč. slova correlative microscopy ; plasma membrane ; nanodomains ; auxin carriers Vědní obor RIV EF - Botanika Obor OECD Plant sciences, botany CEP GA19-03909S GA ČR - Grantová agentura ČR Způsob publikování Open access Institucionální podpora UPT-D - RVO:68081731 UT WOS 000712762200001 EID SCOPUS 85115616208 DOI 10.3390/biom11101407 Anotace Fluorescence light microscopy provided convincing evidence for the domain organization of plant plasma membrane (PM) proteins. Both peripheral and integral PM proteins show an inhomo-geneous distribution within the PM. However, the size of PM nanodomains and protein clusters is too small to accurately determine their dimensions and nano-organization using routine confocal fluorescence microscopy and super-resolution methods. To overcome this limitation, we have developed a novel correlative light electron microscopy method (CLEM) using total internal reflection fluorescence microscopy (TIRFM) and advanced environmental scanning electron microscopy (A-ESEM). Using this technique, we determined the number of auxin efflux carriers from the PINFORMED (PIN) family (NtPIN3b-GFP) within PM nanodomains of tobacco cell PM ghosts. Protoplasts were attached to coverslips and immunostained with anti-GFP primary antibody and secondary antibody conjugated to fluorochrome and gold nanoparticles. After imaging the nanodomains within the PM with TIRFM, the samples were imaged with A-ESEM without further processing, and quantification of the average number of molecules within the nanodomain was performed. Without requiring any post-fixation and coating procedures, this method allows to study details of the organization of auxin carriers and other plant PM proteins. Pracoviště Ústav přístrojové techniky Kontakt Martina Šillerová, sillerova@ISIBrno.Cz, Tel.: 541 514 178 Rok sběru 2022 Elektronická adresa https://www.mdpi.com/2218-273X/11/10/1407
Počet záznamů: 1