"MS-Patch-Clamp" or the Possibility of Mass Spectrometry Hybridization with Patch-Clamp Setups for Single Cell Metabolomics and Channelomics

Abstract

In this projecting work we propose a mass spectrometric patch-clamp equipment with the capillary performing both a local potential registration at the cell membrane and the analyte suction simultaneously. This paper provides a current literature analysis comparing the possibilities of the novel approach proposed with the known methods, such as scanning patch-clamp, scanning ion conductance microscopy, patch clamp based on scanning probe microscopy technology, quantitative subcellular secondary ion mass spectrometry or "ion microscopy", live single-cell mass spectrometry, in situ cell-by-cell imaging, single-cell video-mass spectrometry, etc. We also consider the ways to improve the informativeness of these methods and particularly emphasize the trend at the increasing of the analysis complexity. We propose here the way to improve the efficiency of the cell trapping to the capillary during MS-path-clamp, as well as to provide laser surface ionization using laser trapping and tweezing of cells with the laser beam transmitted through the capillary as a waveguide. It is also possible to combine the above system with the microcolumn separation system or capillary electrophoresis as an optional direction of further development of the complex of analytical techniques emerging from the MS variation of patch-clamp.

Single‐Cell Mass Spectrometry Approaches to Explore Cellular Heterogeneity

“…To achieve multifaceted analysis capturing additional cellular properties and obtain more dynamic information, single-cell MS can be coupled with other analytical tools to explore cell behavior and specific subpopulations.T hese techniques are based on non-destructive approaches that allow subsequent MS analysis.T hey include patch-clamp experiments, [135] fluorescence microscopy, [7,8] and Raman spectroscopy. [9] Theavailability of high-throughput cell sorting and singlecell RNA-seq combined with the emerging single-cell proteomics and metabolomics methods raises the prospect of single-cell systems biology.Since the studied cells come from awell-defined subpopulation, the underlying biological pathways are better defined and the corresponding bioinformatics can be simplified.…”

Simultaneous <i>in situ</i> Detection of the Optical Fluorescence, Fluorescence Recovery Kinetics After Photobleaching & Membrane Ion Flux on the Electrophysiological Lab-on-a-Chip

“…The above concept gave rise to the experiments on the patch clamp of the synthetic ionic channels and artificial cells, as well as to the hybridization of the real-time patch clamp with the mass spectrometric monitoring (the so-called MS-patch-clamp) [24], which are the continuation of the work initiated by the authors in the early 2000-th.…”

Isotopic Mass-Dependent MS-Patch-Clamp and Isotopic Mass-Independent ESR-Patch-Clamp

“…This conclusion is not applicable for multi-isotope mass-spectrometry, including multi-isotope mass-spectrometric imaging [90][91][92][93][94][95][96][97][98] using liquid pumping and adjustable capillary systems similar to those used in the standard patch-clamp configuration [99][100]. Earlier we have already shown the possibility of the MS-patch-clamp techniques [101,102], but those theoretical considerations fail to provide reliable results without strong biochemical argumentation in the frame of the biophysics of the isotope-labeled agents, such as receptors, antagonists, blockers, transmitters, etc. From the above considerations one can conclude that selective amplification of the isotope uptake by the electrical diffusion potentials is a direct consequent of the ion channel-mediated flux occurring in the membrane vesicles [103] and the isotope flows and flux ratios in biological membranes, which are interrelated [104] as in the ion exchange membranes [105][106].…”