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No Need for Labels: The Autofluorescence of Leishmania tarentolae Mitochondria and the Necessity of Negative Controls

BACKGROUND: Fluorescence microscopy is a powerful tool to study the morphology and function of subcellular compartments or to determine the localization of proteins. The method is also regularly used for the analysis of parasitic protists including kinetoplastida. RESULTS: Here, we report a signific...

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Detalles Bibliográficos
Autores principales: Eckers, Elisabeth, Deponte, Marcel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3471854/
https://www.ncbi.nlm.nih.gov/pubmed/23077654
http://dx.doi.org/10.1371/journal.pone.0047641
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author Eckers, Elisabeth
Deponte, Marcel
author_facet Eckers, Elisabeth
Deponte, Marcel
author_sort Eckers, Elisabeth
collection PubMed
description BACKGROUND: Fluorescence microscopy is a powerful tool to study the morphology and function of subcellular compartments or to determine the localization of proteins. The method is also regularly used for the analysis of parasitic protists including kinetoplastida. RESULTS: Here, we report a significant autofluorescence of Leishmania tarentolae mitochondria. The autofluorescence, presumably caused by flavoproteins, was detectable using a variety of cell fixation protocols and had a maximum emission at approximately 538 nm. Stable signals were obtained with xenon lamps as a light source and filter sets that are commonly used for the detection of green fluorescent protein. CONCLUSIONS: On the one hand, we present a methodological approach to examine mitochondrial morphology or to study the colocalization of mitochondrial proteins without additional staining or labeling procedures. On the other hand, under certain experimental conditions, mitochondrial autofluorescence can result in false positive signals, demonstrating the necessity to analyze unlabeled cells as negative controls.
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spelling pubmed-34718542012-10-17 No Need for Labels: The Autofluorescence of Leishmania tarentolae Mitochondria and the Necessity of Negative Controls Eckers, Elisabeth Deponte, Marcel PLoS One Research Article BACKGROUND: Fluorescence microscopy is a powerful tool to study the morphology and function of subcellular compartments or to determine the localization of proteins. The method is also regularly used for the analysis of parasitic protists including kinetoplastida. RESULTS: Here, we report a significant autofluorescence of Leishmania tarentolae mitochondria. The autofluorescence, presumably caused by flavoproteins, was detectable using a variety of cell fixation protocols and had a maximum emission at approximately 538 nm. Stable signals were obtained with xenon lamps as a light source and filter sets that are commonly used for the detection of green fluorescent protein. CONCLUSIONS: On the one hand, we present a methodological approach to examine mitochondrial morphology or to study the colocalization of mitochondrial proteins without additional staining or labeling procedures. On the other hand, under certain experimental conditions, mitochondrial autofluorescence can result in false positive signals, demonstrating the necessity to analyze unlabeled cells as negative controls. Public Library of Science 2012-10-15 /pmc/articles/PMC3471854/ /pubmed/23077654 http://dx.doi.org/10.1371/journal.pone.0047641 Text en © 2012 Eckers, Deponte http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Eckers, Elisabeth
Deponte, Marcel
No Need for Labels: The Autofluorescence of Leishmania tarentolae Mitochondria and the Necessity of Negative Controls
title No Need for Labels: The Autofluorescence of Leishmania tarentolae Mitochondria and the Necessity of Negative Controls
title_full No Need for Labels: The Autofluorescence of Leishmania tarentolae Mitochondria and the Necessity of Negative Controls
title_fullStr No Need for Labels: The Autofluorescence of Leishmania tarentolae Mitochondria and the Necessity of Negative Controls
title_full_unstemmed No Need for Labels: The Autofluorescence of Leishmania tarentolae Mitochondria and the Necessity of Negative Controls
title_short No Need for Labels: The Autofluorescence of Leishmania tarentolae Mitochondria and the Necessity of Negative Controls
title_sort no need for labels: the autofluorescence of leishmania tarentolae mitochondria and the necessity of negative controls
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3471854/
https://www.ncbi.nlm.nih.gov/pubmed/23077654
http://dx.doi.org/10.1371/journal.pone.0047641
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