Publication details

Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells

Authors

LEDVINA Vojtěch JANEČKOVÁ Eva MATALOVÁ Eva KLEPÁRNÍK Karel

Year of publication 2017
Type Article in Periodical
Magazine / Source Analytical and Bioanalytical chemistry
MU Faculty or unit

Faculty of Science

Citation
web Full Text
Doi http://dx.doi.org/10.1007/s00216-016-9998-6
Keywords Single-cell analysis; Bioluminescence; Apoptosis; Caspase-3/7
Description 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(A (R)) 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.

You are running an old browser version. We recommend updating your browser to its latest version.

More info