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A miniaturized device for bioluminescence analysis of caspase-3/7 activity in a single apoptotic cell
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SYSNO ASEP 0437556 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název A miniaturized device for bioluminescence analysis of caspase-3/7 activity in a single apoptotic cell Tvůrce(i) Adamová, Eva (UZFG-Y)
Lišková, M. (CZ)
Matalová, Eva (UZFG-Y) RID
Klepárník, K. (CZ)Zdroj.dok. Analytical and Bioanalytical Chemistry. - : Springer - ISSN 1618-2642
Roč. 406, č. 22 (2014), s. 5389-5394Poč.str. 6 s. Forma vydání Tištěná - P Jazyk dok. eng - angličtina Země vyd. DE - Německo Klíč. slova apoptosis ; bioluminiscence ; caspase3/7 ; single-cell analysis Vědní obor RIV CE - Biochemie CEP GAP304/11/1418 GA ČR - Grantová agentura ČR Institucionální podpora UZFG-Y - RVO:67985904 UT WOS 000340877200017 EID SCOPUS 84907600555 DOI 10.1007/s00216-014-7949-7 Anotace Caspases are key enzymes activated during the apoptotic machinery. Apoptosis as a way of programmed cell death becomes deregulated in some pathologies including cancer transformations, neurodegenerative, or autoimmune diseases. Most of the methods available for the detection of apoptosis and caspases provide qualitative information only or quantification data as an average from cell populations or cell lysates. Several reports point to the importance of more accurate single-cell analyses in biomedical studies due to heterogeneity at tissue as well as cell level. To meet these requirements, we developed a miniaturized device enabling detection and quantification of active caspase-3/7 in individual cells at a femtogram level (10(-15) g). The active caspase-3/7 detection protocol is based on the bioluminescence chemistry commercially available as a Caspase-Glo (TM) 3/7 reagent developed by Promega. As a model, we used human stem cells treated by camptothecin to induce apoptosis. Individual apoptotic cells were captured from a culture medium under a microscope and transferred by a micromanipulation system into a detection capillary containing 2 mu l of the reagent. Cells without activation by camptothecin served as negative controls. The detection limit of active caspase-3/7 achieved in the miniaturized system was determined as 0.20 and limit of quantification as 0.65 of the amount found in a single apoptotic human stem cell. Such a sensitive method could have a wide application potential in laboratory medicine and related clinically oriented research. Pracoviště Ústav živočišné fyziologie a genetiky Kontakt Jana Zásmětová, knihovna@iapg.cas.cz, Tel.: 315 639 554 Rok sběru 2015
Počet záznamů: 1