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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F17%3A00108746" target="_blank" >RIV/00216224:14310/17:00108746 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/68081715:_____/17:00463697 RIV/67985904:_____/17:00463697 RIV/62157124:16170/17:43875773

Výsledek na webu

<a href="https://link.springer.com/content/pdf/10.1007%2Fs00216-016-9998-6.pdf" target="_blank" >https://link.springer.com/content/pdf/10.1007%2Fs00216-016-9998-6.pdf</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s00216-016-9998-6" target="_blank" >10.1007/s00216-016-9998-6</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Analytical and Bioanalytical chemistry

ISSN

1618-2642

e-ISSN

1618-2650

Svazek periodika

409

Číslo periodika v rámci svazku

1

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

6

Strana od-do

269-274

Kód UT WoS článku

000391357700027

EID výsledku v databázi Scopus

2-s2.0-84991710989