Capillary Electrophoresis Strategy to Monitor the Released and Remaining Nitric Oxide from the Same Single Cell Using a Specially Designed Water-Soluble Fluorescent Probe

Abstract

Gasotransmitters including nitric oxide (NO), carbon monoxide (CO), and hydrogen sulfide (H2S) have attracted more and more attention in the past decades due to their unique signaling and functions. However, as a fundamental issue in the investigations of gasotransmitters, the cell membrane permeability and release behavior of them is controversial in reports because of the lack of an efficient approach to determine gasotransmitters released out of and remaining in the same cells simultaneously. To solve such problem, taking NO as representative, a robust and facile strategy has been reported based on a completely water-soluble fluorescent probe and a commercially available capillary electrophoresis system. A specially designed boron-dipyrromethene (BODIPY)-based fluorescent probe with two sulfonate groups, disodium 2,6-disulfonate-1,3,5,7-tetramethyl-8-(3',4'-diaminophenyl) difluoroboradiaza-s-indance (TMDSDAB), has been developed. As a turn-on fluorescent probe, TMDSDAB can react with NO promptly in aqueous media, and 540-fold enhancement of fluorescence is obtained. Using TMDSDAB, the trapping and quantification of NO released out of and remaining in the same single cell was achieved by capillary electrophoresis with laser-induced fluorescence detection. The limit of detection is 0.5 nM for NO. The proposed method has been applied to estimate the release behavior of single macrophages, and the results indicated that the cell membrane should be a barrier to NO diffusion.

Design and Application of Sensors for Chemical Cytometry

“…Fluorescence detection is typically performed on-column by interrogating the capillary with a focused light source, generally a laser, and detecting emitted light with a photomultiplier tube (PMT) or photodiode. 18, 21, 26, 45 ED coupled to CE offers a multitude of advantages for single cell analysis due to its high sensitivity, low LOD, and low cost. 23, 25, 49, 50 MS coupled to CE has led to impressive advances in elucidation of the proteome and metabolome of single cells.…”

“…created the water soluble diamino BODIPY probe 3 by adding sulfonate groups so that both extracellular and intracellular NO ( 4 ) of a trapped cell can be measured before and after lysis. 79…”

Recent Advances in Plasmonic Nanostructures Applied for Label‐free Single‐cell Analysis

“…[2]. Up to date, numerous analytical techniques have been developed for single‐cell analysis, such as capillary electrophoresis [3], mass spectrometry [4], fluorescence [5], DNA sequencing [6], western blotting [7], and electrochemical detection [8]. Although these popular methods have played important roles in heterogeneity analysis at single‐cell level, the procedures of cell lysis and fluorophore labeling complicate the analysis process, increasing the possibility of false signals caused by photobleaching.…”

Water-soluble Hantzsch ester as switch-on fluorescent probe for efficiently detecting nitric oxide

“…Up to date, the approaches involved in the NO detection cover electron paramagnetic resonance spectroscopy [11], colorimetric method [12], electrochemical method [13], chemiluminescence [14], and fluorescence [15][16][17]. Among those developed analytical methods, fluorescence showed tremendous advantages when it comes to high sensitivity and selectivity [18][19][20]. Additionally, fluorescence can also be used in both solutions and solid phase, and even gas phase.…”