Novel dye for detection of callus embryo by confocal laser scanning fluorescence microscopy

• Published in: Luminescence (Chichester, England) Vol. 34; no. 3; pp. 353 - 359
• Main Authors: Kirilova, Elena, Mickevica, Ilona, Mezaraupe, Ligita, Puckins, Aleksandrs, Rubenina, Ilze, Osipovs, Sergejs, Kokina, Inese, Bulanovs, Andrejs, Kirjusina, Muza, Gavarane, Inese
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01.05.2019
• Subjects: Alfalfa; Animal embryos; Benz(a)Anthracenes - chemistry; benzanthrone dye; Callus; Cell culture; confocal laser scanning fluorescence microscopy; Detection; Dyes; embryo; Embryos; Flax; Flax - chemistry; Flax - embryology; Fluorescence; Fluorescence microscopy; Fluorescent dyes; Fluorescent Dyes - chemistry; Fluorescent indicators; Lasers; Medicago sativa - chemistry; Medicago sativa - embryology; Microscopy; Microscopy, Confocal - methods; Organic solvents; Red clover; Specificity; Synthesis; Tissue Culture Techniques; Tissues; Trifolium - chemistry; Trifolium - embryology; Visualization; benzanthrone dye; embryo; callus; confocal laser scanning fluorescence microscopy
• ISSN: 1522-7235; 1522-7243
• DOI: 10.1002/bio.3616

In the present study a new luminescent dye 3‐N‐(2‐pyrrolidinylacetamido)benzanthrone (AZR) was synthesized. Spectroscopic measurements of the novel benzanthrone 3‐aminoderivative were performed in seven organic solvents showing strong fluorescence. The capability of the prepared dye for visualization has been tested on flax, red clover and alfalfa to determinate the embryo in plant callus tissue cultures. Callus cells were stained with AZR and further analysed utilizing confocal laser scanning fluorescence microscopy. Performed experiments show high visualization effectiveness of newly synthesized fluorescent dye AZR that is efficient in fast and relatively inexpensive diagnostics of callus embryos that are problematic due to in vitro culture specificity.