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Immunocytochemical Visualization of Proteins from Cyanobacterial Cells with High Autofluorescence of Phycoerythrin and Phycourobilin

The result's identifiers

  • Result code in IS VaVaI

    <a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F23%3A00582456" target="_blank" >RIV/61388971:_____/23:00582456 - isvavai.cz</a>

  • Result on the web

    <a href="https://www.jove.com/t/65168/immunocytochemical-visualization-proteins-from-cyanobacterial-cells" target="_blank" >https://www.jove.com/t/65168/immunocytochemical-visualization-proteins-from-cyanobacterial-cells</a>

  • DOI - Digital Object Identifier

    <a href="http://dx.doi.org/10.3791/65168" target="_blank" >10.3791/65168</a>

Alternative languages

  • Result language

    angličtina

  • Original language name

    Immunocytochemical Visualization of Proteins from Cyanobacterial Cells with High Autofluorescence of Phycoerythrin and Phycourobilin

  • Original language description

    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.

  • Czech name

  • Czech description

Classification

  • Type

    J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database

  • CEP classification

  • OECD FORD branch

    10606 - Microbiology

Result continuities

  • Project

    <a href="/en/project/GA20-17627S" target="_blank" >GA20-17627S: C and N metabolisms and their impact on ecological significance of unicellular diazotrophic cyanobacteria</a><br>

  • Continuities

    I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Others

  • Publication year

    2023

  • Confidentiality

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

Data specific for result type

  • Name of the periodical

    Jove-Journal of Visualized Experiments

  • ISSN

    1940-087X

  • e-ISSN

  • Volume of the periodical

    2023

  • Issue of the periodical within the volume

    199

  • Country of publishing house

    US - UNITED STATES

  • Number of pages

    16

  • Pages from-to

    e65168

  • UT code for WoS article

    001159858200019

  • EID of the result in the Scopus database

    2-s2.0-85172771682