The main difference between non-damage micro-measurement technology and patch clamp technology

The birth of patch clamp technology in 1976 was an important event in the history of modern life science research. Two German scientists won the 1991 Nobel Prize in Physiology or Medicine for their achievements in ion channel research using patch clamp technology. The research of patch clamp technology on the opening and closing of ion channels has become an important bridge connecting biomolecules and biological functions, which has led to a large number of high-level research results.

However, with the wide application of patch clamp technology, many of its inherent problems are gradually exposed. First of all, the patch clamp technique measurement needs to be realized by the process of adsorbing cell membrane by microelectrode, which is extremely difficult to operate, requires long training by the experimenter, and also severely limits the range of the patch clamp technique to detect the sample. For measuring biological cells.

The study of ion channel by patch clamp has the incomparable advantages of other techniques, but the patch clamp technique records the current. For the study of ion transmembrane transport, only recording current may cause information loss. The study found that in addition to the ion channel (Ion Channel) of ions, there is also a transporter model. The mere study of ion channels does not reflect all the information of ion transport. On the one hand, the transport process of ions through the carrier tends to be slow, the current generated is very weak, and the patch clamp technique is difficult to record; more importantly, if the ions are transported "into one out" or "one in one out", It will cause overall electrical neutrality, no current, and cannot be recorded by patch clamp technology. At the same time, patch clamp technology can't do anything about the transport of neutral molecules.

In addition, the study of ion channel opening and closing by patch clamp technique to characterize the biological function process is too indirect for many fields, and the severe damage to the cell by the film-clamping process of the patch-clamp experiment may seriously affect the measured data. Problems such as authenticity have become the focus of the patch clamp technique.

With the development of life sciences, especially the study of biological functions and physiological mechanisms, it has gradually become the mainstream of life science research. The majority of science and technology workers urgently need a more comprehensive, direct and convenient ion molecular information representation technology.

In 1990, in the famous Marine Biological Laboratory (MBL) in the United States, non-invasive micro-testing technology came into being. The non-invasive micro-test technique uses an ion/molecular-selective microelectrode (this microelectrode is dedicated to non-damage micro-testing techniques, unlike patch clamps or microelectrodes used in other techniques) to measure near the sample rather than in a non-invasive or non-invasive manner. Obtain values ​​of ion molecule concentration, flow rate, and flow direction into and out of the sample surface. Due to its unique non-invasive measurement method, non-invasive micro-measurement technology can measure a wide range of samples, from biological whole body, organ, tissue, cell layer, single cell to organelle.

The ion molecular flow rate and flow direction information obtained by non-invasive micro-testing technology not only reflect the dynamic process of biophysical activity, but also the relative overall information. For example, the cell ion molecular flow rate information reflects several ion channels on the cell membrane. As a result of interaction with the ionophore or several molecular transport processes, the tissue ion molecular flow rate information reflects the interaction of several cells on the tissue, independent of the ion transport mode and whether the transport process is electrically neutral. This makes the ion molecular flow information obtained by non-invasive micro-testing technology a very comprehensive and direct means of characterizing biological functions and physiological mechanisms.

The experimental operation of the non-invasive micro-measurement technology is mainly micromanipulation, and does not involve complicated operation processes such as adsorption of cell membranes. It is very convenient and quick, and the experimenter can get started after simple training. In addition, non-damage micro-measurement technology has the advantages that long-term, multi-electrode, arbitrary path measurement and other technologies are incomparable.

The many advantages and wide applicability of non-invasive micro-test technology have made it more and more widely used since its birth, providing a new perspective and ideas for life science research. Scholars from many different fields have applied non-invasive micro-testing technology to carry out research work. More than 100 high-level research papers have been published in famous journals such as Nature, Nature Protocols, PNAS and Plant Cell. In addition to the field of life sciences, non-invasive micro-testing technology has also attracted the interest of science and technology workers in other disciplines such as materials science, and has also achieved fruitful results in the research of these disciplines.

Traditional physiologists with a focus on ion channel research, although patch-clamp technology is their primary research tool, non-invasive micro-testing techniques can also make their research deeper and fuller. For example, Australian scholar Shabala et al. published a research paper published in "Plant Physiology" in 2006, using Arabidopsis thaliana as a material to study the effect of extracellular Ca ²⁺ on NaCl-induced K ⁺ loss. The authors used non-invasive microassay to measure the K ⁺ flow in Arabidopsis roots and leaves, and found that the increase of extracellular Ca ²⁺ concentration can effectively inhibit NaCl-induced K ⁺ loss; patch clamp measurement of Arabidopsis root protoplasts It was found that extracellular Ca ²⁺ can affect the permeability of K ⁺ channels on the cell membrane. These work have made the research results of this paper very full, and clearly explained the plant-related physiological mechanisms from different levels of cells to organs.

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