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
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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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