Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells
Language English Country Germany Media print-electronic
Document type Journal Article
PubMed
27757513
DOI
10.1007/s00216-016-9998-6
PII: 10.1007/s00216-016-9998-6
Knihovny.cz E-resources
- Keywords
- Apoptosis, Bioluminescence, Caspase-3/7, Single-cell analysis,
- MeSH
- Single-Cell Analysis instrumentation methods MeSH
- Apoptosis * MeSH
- Equipment Design MeSH
- Enzyme Assays instrumentation methods MeSH
- Caspase 3 analysis metabolism MeSH
- Caspase 7 analysis metabolism MeSH
- Cells, Cultured MeSH
- Luminescent Measurements instrumentation methods MeSH
- Mice MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Caspase 3 MeSH
- Caspase 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.
References provided by Crossref.org
Caspase-8 Deficient Osteoblastic Cells Display Alterations in Non-Apoptotic Pathways
Osteogenic impact of pro-apoptotic caspase inhibitors in MC3T3-E1 cells
