Počet záznamů: 1
Immunocytochemical Visualization of Proteins from Cyanobacterial Cells with High Autofluorescence of Phycoerythrin and Phycourobilin
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SYSNO ASEP 0582456 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Immunocytochemical Visualization of Proteins from Cyanobacterial Cells with High Autofluorescence of Phycoerythrin and Phycourobilin Tvůrce(i) Masuda, Takako (MBU-M) ORCID
Majerová, Dominika (MBU-M)
Piwosz, Kasia (MBU-M) ORCID
Tsurumaki, Tatsuhiro (MBU-M)
Fujita, Y. (JP)
Prášil, Ondřej (MBU-M) RID, ORCIDČíslo článku e65168 Zdroj.dok. Jove-Journal of Visualized Experiments - ISSN 1940-087X
Roč. 2023, č. 199 (2023)Poč.str. 16 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova Chlorophyl ; phycoerythrin ; cyanobacterium ; phycourobilin Obor OECD Microbiology CEP GA20-17627S GA ČR - Grantová agentura ČR Způsob publikování Omezený přístup Institucionální podpora MBU-M - RVO:61388971 UT WOS 001159858200019 EID SCOPUS 85172771682 DOI 10.3791/65168 Anotace Presented is a protocol for visualizing and quantifying a specific protein in cells at the cellular level for the marine cyanobacterium Crocosphaera watsonii, a crucial primary producer and nitrogen fixer in oligotrophic oceans. One of the challenges for marine autotrophic N2 fixers (diazotrophs) is distinguishing probe-derived fluorescence signals from autofluorescence. C. watsonii was selected to represent chlorophyll-, phycoerythrin-and phycourobilin-containing cyanobacteria. The protocol allows for simple and semi-quantitative visualization of proteins in C. watsonii at a single-cell level, enabling investigation of protein production under different environmental conditions to evaluate the metabolic activities of the target cyanobacteria. Furthermore, the fixation and permeabilization methods are optimized to enhance the fluorescence signals from target proteins to distinguish them from autofluorescence, especially from phycoerythrin and phycourobilin. The enhanced signal can be visualized using confocal or widefield fluorescence microscopy. Additionally, fluorescence intensity was semi-quantified using Fiji software. This single-cell analysis workflow allows the evaluation of cell-to-cell variations of specific protein content. The protocol can be performed in any life science laboratory as it requires only standard equipment and can also be easily adapted to other phycoerythrin-containing cyanobacterial cells. © 2023 JoVE Journal of Visualized Experiments. Pracoviště Mikrobiologický ústav Kontakt Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Rok sběru 2024 Elektronická adresa https://www.jove.com/t/65168/immunocytochemical-visualization-proteins-from-cyanobacterial-cells
Počet záznamů: 1