Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells
Jazyk angličtina Země Německo Médium print-electronic
Typ dokumentu časopisecké články
PubMed
27757513
DOI
10.1007/s00216-016-9998-6
PII: 10.1007/s00216-016-9998-6
Knihovny.cz E-zdroje
- Klíčová slova
- Apoptosis, Bioluminescence, Caspase-3/7, Single-cell analysis,
- MeSH
- analýza jednotlivých buněk přístrojové vybavení metody MeSH
- apoptóza * MeSH
- design vybavení MeSH
- enzymatické testy přístrojové vybavení metody MeSH
- kaspasa 3 analýza metabolismus MeSH
- kaspasa 7 analýza metabolismus MeSH
- kultivované buňky MeSH
- luminiscenční měření přístrojové vybavení metody MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kaspasa 3 MeSH
- kaspasa 7 MeSH
Analysing the chemical content of individual cells has already been proven to reveal unique information on various biological processes. Single-cell analysis provides more accurate and reliable results for biology and medicine than analyses of extracts from cell populations, where a natural heterogeneity is averaged. To meet the requirements in the research of important biologically active molecules, such as caspases, we have developed a miniaturized device for simultaneous analyses of individual cells. A stainless steel body with a carousel holder enables high-sensitivity parallel detections in eight microvials. The holder is mounted in front of a photomultiplier tube with cooled photocathode working in photon counting mode. The detection of active caspase-3/7, central effector caspases in apoptosis, in single cells is based on the bioluminescence chemistry commercially available as Caspase-Glo® 3/7 reagent developed by Promega. Individual cells were captured from a culture medium under microscope and transferred by micromanipulator into detection microvial filled with the reagent. As a result of testing, the limits of detection and quantification were determined to be 0.27/0.86 of active caspase-3/7 content in an average apoptotic cell and 0.46/2.92 for non-apoptotic cells. Application potential of this technology in laboratory diagnostics and related medical research is discussed. Graphical abstract Miniaturized device for simultaneous analyses of individual cells.
Citace poskytuje Crossref.org
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