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

J 2017

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

LEDVINA, Vojtěch, Eva JANEČKOVÁ, Eva MATALOVÁ a Karel KLEPÁRNÍK

Základní údaje

Originální název

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

Autoři

LEDVINA, Vojtěch (203 Česká republika, domácí), Eva JANEČKOVÁ (203 Česká republika, domácí), Eva MATALOVÁ (203 Česká republika) a Karel KLEPÁRNÍK (203 Česká republika, garant)

Vydání

Analytical and Bioanalytical chemistry, HEIDELBERG, SPRINGER HEIDELBERG, 2017, 1618-2642

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Odkazy

Full Text

Impakt faktor

Impact factor: 3.307

Kód RIV

RIV/00216224:14310/17:00108746

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1007/s00216-016-9998-6

UT WoS

000391357700027

Klíčová slova anglicky

Single-cell analysis; Bioluminescence; Apoptosis; Caspase-3/7

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 17. 12. 2019 09:09, Mgr. Marie Šípková, DiS.

Anotace

V originále

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.

Citovat

LEDVINA, Vojtěch, Eva JANEČKOVÁ, Eva MATALOVÁ a Karel KLEPÁRNÍK. Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells. Analytical and Bioanalytical chemistry. HEIDELBERG: SPRINGER HEIDELBERG, 2017, roč. 409, č. 1, s. 269-274. ISSN 1618-2642. Dostupné z: https://dx.doi.org/10.1007/s00216-016-9998-6.

@article{1387810,
   author = {Ledvina, Vojtěch and Janečková, Eva and Matalová, Eva and Klepárník, Karel},
   article_location = {HEIDELBERG},
   article_number = {1},
   doi = {http://dx.doi.org/10.1007/s00216-016-9998-6},
   keywords = {Single-cell analysis; Bioluminescence; Apoptosis; Caspase-3/7},
   language = {eng},
   issn = {1618-2642},
   journal = {Analytical and Bioanalytical chemistry},
   title = {Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells},
   url = {https://link.springer.com/content/pdf/10.1007%2Fs00216-016-9998-6.pdf},
   volume = {409},
   year = {2017}
}

TY  - JOUR
ID  - 1387810
AU  - Ledvina, Vojtěch - Janečková, Eva - Matalová, Eva - Klepárník, Karel
PY  - 2017
TI  - Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells
JF  - Analytical and Bioanalytical chemistry
VL  - 409
IS  - 1
SP  - 269-274
EP  - 269-274
PB  - SPRINGER HEIDELBERG
SN  - 16182642
KW  - Single-cell analysis
KW  - Bioluminescence
KW  - Apoptosis
KW  - Caspase-3/7
UR  - https://link.springer.com/content/pdf/10.1007%2Fs00216-016-9998-6.pdf
L2  - https://link.springer.com/content/pdf/10.1007%2Fs00216-016-9998-6.pdf
N2  - 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.
ER  -

LEDVINA, Vojtěch, Eva JANEČKOVÁ, Eva MATALOVÁ a Karel KLEPÁRNÍK. Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells. \textit{Analytical and Bioanalytical chemistry}. HEIDELBERG: SPRINGER HEIDELBERG, 2017, roč.~409, č.~1, s.~269-274. ISSN~1618-2642. Dostupné z: https://dx.doi.org/10.1007/s00216-016-9998-6.

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