Capillary zone electrophoresis‐based cytotoxicity analysis of Caco‐2 cells

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.

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“…Besides, borate buffer has been used in the electrophoresis for cells. 30 Therefore, borate buffer is chosen as the background electrolyte.…”

Section: ■ Results and Discussionmentioning

confidence: 80%

“…But in borate buffer, RAW 264.7 macrophage can live for at least 30 min, and its current is low enough. Besides, borate buffer has been used in the electrophoresis for cells . Therefore, borate buffer is chosen as the background electrolyte.…”

Section: Resultsmentioning

confidence: 99%

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

et al. 2015

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“…EP of cells is commonly conducted under a uniform direct current (DC) electric field and has long been used for cell researches to have insights into the cellular membrane property and the interactions of cells with molecules or other cells . Cellular membranes are known to consist of numerous chargeable proteins, lipids, teichoic acids, glycocalyx, and lipopolisaccharides, which endow the whole cells with characteristic and measurable electrophoretic mobility (μ EP ) and can thus be determined by electrophoresis .…”

Section: Introductionmentioning

confidence: 99%

Simultaneous determination of electrophoretic and dielectrophoretic mobilities of human red blood cells

Xiu-zhen

Chen

2015

Electrophoresis

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Electrophoresis and dielectrophoresis of cells can reveal many distinct cellular properties but are often conducted separately. Herein a simultaneous strategy was proposed, and a simple method was established by making cells migrate through a cross channel under a micro video for real-time observation. The experiment can be performed within 0.044-1 s. In combination with digital calculation based on electromagnetic theory, the method was validated to be applicable to the determination of electrophoretic and dielectrophoretic mobilities, μEP and μDEP , of human blood erythrocytes, giving μEP = -(0.87 ± 0.16)× 10(-4) cm(2) ·V(-1) · s(-1) and μDEP = -(4.5 ± 1.3) × 10(-8) cm(4) ·V(-2) ·s(-1) by vector decomposition, or μEP = -(0.89 ± 0.14) × 10(-4) cm(2) ·V(-1) · s(-1) and μDEP = -(4.6 ±1.2) × 10(-8) cm(4) ·V(-2) · s(-1) by least squares fitting, all agreeing with published data. Hydrodynamic and EOFs were eliminated for better measurement. It was found that the location of cells had a serious impact on the measurement precision, and the upstream of the cross channel along the electric field was chosen for precise measurement. The method is also extendable to the study of other cells and particles.

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Analyses of anticancer drugs by capillary electrophoresis: a review

Ali

Haque

Wani

et al. 2013

Biomedical Chromatography

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Capillary electrophoresis is a fast, inexpensive and low detection limit technique for the analysis of anticancer drugs. It has been used to analyze various anticancer drugs in biological samples, pharmaceutical preparations and environmental matrices. It has also been used to detect various cancer biomarkers in cancer patients. The present article describes the state-of-the art of capillary electrophoresis for the analyses of anticancer drugs. Various drugs discussed belong to several groups such as antimitotic agents, nucleoside analogs, antibiotics, topoisomerase inhibitors and DNA intercalating agents. In addition, efforts have also been made to discuss sample preparation, applications of capillary electrophoresis in genomic research, optimization and future perspectives.

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Capillary zone electrophoresis‐based cytotoxicity analysis of Caco‐2 cells

Cited by 5 publications

References 33 publications

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

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

Simultaneous determination of electrophoretic and dielectrophoretic mobilities of human red blood cells

Analyses of anticancer drugs by capillary electrophoresis: a review

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