Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells.
Anal Bioanal Chem
; 409(1): 269-274, 2017 Jan.
Article
en En
| MEDLINE
| ID: mdl-27757513
ABSTRACT
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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Apoptosis
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Caspasa 3
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Caspasa 7
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Pruebas de Enzimas
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Análisis de la Célula Individual
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Mediciones Luminiscentes
Límite:
Animals
Idioma:
En
Revista:
Anal Bioanal Chem
Año:
2017
Tipo del documento:
Article
País de afiliación:
República Checa