Detailed Information on Publication Record
2006
Benzophenanthridine alkaloids as a new supravital DNA dyes.
SLANINOVÁ, Iva, Jiří ŠINKORA and Eva TÁBORSKÁBasic information
Original name
Benzophenanthridine alkaloids as a new supravital DNA dyes.
Name in Czech
Benzofenantridinové alkaloidy - nové supravitální DNA barviva.
Authors
SLANINOVÁ, Iva, Jiří ŠINKORA and Eva TÁBORSKÁ
Edition
Praha, Sborník XIV. Cytoskeletálního klubu, p. 20-20, 1 pp. 2006
Publisher
Československá biologická společnost
Other information
Language
English
Type of outcome
Stať ve sborníku
Field of Study
Genetics and molecular biology
Country of publisher
Czech Republic
Confidentiality degree
není předmětem státního či obchodního tajemství
Organization unit
Faculty of Medicine
Keywords in English
benzophenanthridine alkaloids; DNA dye; flow cytometry; fluorescence microscopy; multicolor analysis; cell cycle
Tags
Tags
International impact
Změněno: 12/12/2006 11:40, prof. MUDr. Iva Slaninová, Ph.D.
V originále
We have proved that macarpine (MA) and chelirubine (CHR) can be used as supravital fluorescent DNA probes both in fluorescence microscopy and flow cytometry. MA and CHR are quaternary benzo[c]phenanthridine alkaloids (QBA), that are naturally occurring products with noteworthy antiproliferative activity and fluorescence characteristics. QBA are commonly isolated from plants of Fumariaceae, Papaveraceae, Ranunculaceae and Rutaceae families. When added to intact cell suspensions, water stock solutions of the alkaloids (MA, CHR) stain the cells upon brief (in seconds) incubation at low final concentration (0,01-0,001 mg/ml). We have also evaluated the convenience of these minor QBA as supravital nucleic acid probes for time effective multiparameter immunophenotyping of peripheral blood and bone marrow leukocytes. Heparinized peripheral blood or bone marrow aspirate cells were stained on the surface with a combination of fluorochrome-conjuagted monoclonal antibodies directed against surface markers. Then, an appropriate amount of individual QBA was added and, upon dilution, the samples were acquired on a standard (FACSCalibur) or polychromatic (CyAn) flow cytometer. Our results show that spectral characteristics of individual QBA allow for simultaneous surface immunophenotyping and semiquantitative DNA determination. MA and CHR are excitable by common argon lasers (488 nm) emmiting at the range 575 - 750 nm (i.e. fluorescence channels 2-5). Fluorescence channels 1 and 5 – 7 are available for surface immunophenotyping on FACSCalibur and CyAn, respectively.
In Czech
Bylo zjištěno, že benzophenantridinové alkaloidy makarpin, celirubin a sanguinarin jsou využitelné pro značení DNA u živých nepermeabilizovaných buněk. Makarpinu je navích schopen vázat DNA stochiometricky a tím je možné rozlišit fáze buněčného cyklu. Využití makarpinu je ve fluorescenčni mikroskopii pro studium struktury jádra, v prutokové cytometrii - odlišení jaderných buněk i pri mnohobarevném značení v kombinaci se značením jiných znaků např. protilátkami značenými FITC a studium fází buněčného cyklu.
Links
| GA525/04/0017, research and development project |
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| MSM0021622415, plan (intention) |
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