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

Abstract

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.

A Simple and Rapid Staining Technique for Sex Determination of Trichinella Larvae Parasites by Confocal Laser Scanning Microscopy

“…Currently, the synthesis of new high-emitting and low-toxic luminescent dyes (in particular, benzanthrone derivatives) remains an important task for the search of new efficient probes and labels. Recently reported derivatives of 3-acetamidobenzanthrone are highly luminescent compounds and used as sensors for various substances (Wang et al, 2004; Staneva & Betcheva, 2007; Staneva et al, 2019), examination of adult and larvae of trematodes from aquatic animals (Kirilova et al, 2018 a ), and visualization of callus embryo (Kirilova et al, 2019).…”

Comparison of the Benzanthrone Luminophores: They Are Not Equal for Rapid Examination of Parafasciolopsis fasciolaemorpha (Trematoda: Digenea)

“…In literature, there are described various Trematoda stainings methods such as Gower's carmine [24], Ehrlich's acid hematoxylin and Celestin blue-b [13] actin-antibodies and fluorescently labeled phalloidin staining [46][47][48]. Recently we reported that benzanthrone dyes are a useful tool for imaging freshwater trematodes [3], for callus embryo determination [4] and for sex determination of Trichinella larva [49]. Therefore, we continue our research on the synthesis of new benzanthrone markers that bind to actin elements to determine internal organs and systems of trematodes.…”

“…These substances luminescence intensely, have high quantum yields, and their spectral properties are sensitive to the local microenvironment's polarity enabling sensing applications. The spectral properties of the developed fluorescent dyes were investigated in detail [3,4]. As a result, two promising compounds were selected for the current research of the possibilities of studying parasites.…”

“…Studied benzanthrone dyes, using AFA fixative, showed good visualization of internal organ systems and body wall of parasites. Additionally, developed luminescent dye 3-N-(2-pyrrolidinoacetamido) benzanthrone was applicable for callus embryo detection [4]. We can explain the obtained results by specific intermolecular interaction between the applied dye molecules and the stained tissues.…”

3-[N-(4-Methoxybenzyl)amino]benzo[de]anthracen-7-one

“…In connection with our current interest on the synthesis of luminescent compounds for bio-visualization and sensing purposes [5][6][7][8], we report a two-step synthesis of novel substituted amino derivatives of benzanthrone through a condensation reaction between 3-aminobenzanthrone and excess of anisaldehyde to form the azomethine 2, followed by reduction with sodium borohydride in DMF with methanol as a catalyst (at ambient temperature), as depicted in Scheme 1. We decided to carry out the synthesis of imine 2 without solvent because we found that the reaction of initial amine 1 in excess of aldehyde led to the formation of Schiff base 2 in a high yield (80%).…”