SLANINOVÁ, Iva, Jiří ŠINKORA and Eva TÁBORSKÁ. Benzophenanthridine alkaloids as a new supravital DNA dyes. In Sborník XIV. Cytoskeletálního klubu. Praha: Československá biologická společnost. p. 20-20. 2006.
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Basic 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
Original language English
Type of outcome Proceedings paper
Field of Study Genetics and molecular biology
Country of publisher Czech Republic
Confidentiality degree is not subject to a state or trade secret
Organization unit Faculty of Medicine
Keywords in English benzophenanthridine alkaloids; DNA dye; flow cytometry; fluorescence microscopy; multicolor analysis; cell cycle
Tags Benzophenanthridine Alkaloids, cell cycle, DNA dye, flow cytometry, fluorescence microscopy, multicolor analysis
Tags International impact
Changed by Changed by: prof. MUDr. Iva Slaninová, Ph.D., učo 2105. Changed: 12/12/2006 11:40.
Abstract
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.
Abstract (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 projectName: Rostlinné zdroje benzofenanthridinových alkaloidů a účinek těchto alkaloidů na normální a nádorové buňky
Investor: Czech Science Foundation, Plant sources of benzophenanthridine alkaloids and effects of these alkaloid on normal and cancer cells
MSM0021622415, plan (intention)Name: Molekulární podstata buněčných a tkáňových regulací
Investor: Ministry of Education, Youth and Sports of the CR, Molecular basis of cell and tissue regulations
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