Nanopore single-molecule fluorescence imaging device and method under alternating current mode

Abstract

The invention discloses a nano-pore single-molecule fluorescence imaging device and a nano-pore single-molecule fluorescence imaging method under an alternating current mode. The device comprises a nano-pore imaging pool, an alternating current signal providing system and a total internal reflection fluorescence imaging system; an alternating current potential is provided by an alternating currentsignal generating system, and is applied to the two sides of a nano-pore through current amplifying system connecting electrodes. Nano-pore fluorescence imaging is realized through the total internalreflection fluorescence imaging system; the upper side and the lower side of the nano-hole are respectively a low-refractive index medium and a high-refractive index medium, which can realize total internal reflection of exciting light on a nano-pore interface. According to the nano-pore single-molecule fluorescence imaging device and the nano-pore single-molecule fluorescence imaging method in the alternating current mode, the alternating current potential is applied to the two sides of the nano-pore, so that a perforating behaviour of a single molecule can be regulated and controlled; the application of the alternating current potential, the stability of the nano-pore is remarkably improved and the application range of a work voltage is expanded; through the total internal reflection fluorescence imaging system, real-time fluorescence imaging monitoring of the single-molecule signal of the single nano-pore is realized; the nano-pore single-molecule fluorescence imaging device and the nano-pore single-molecule fluorescence imaging method in the alternating current mode have the advantages of high spatial resolution, high time resolution, and high sensitivity.

Description

technical field [0001] The invention relates to the technical field of nanopore single-molecule analysis, in particular to a nanopore single-molecule fluorescence imaging device and method under alternating current mode. Background technique [0002] Nanochannels (pores) widely exist in biomembranes and usually serve the physiological function of transporting biomolecules and ions. They are the basis of material and energy exchange in organisms and play an important role in life processes. The selective transport of substances in biological nanochannels is closely related to a variety of life activities, such as hormone secretion, cardiac beating, formation of transmembrane proton gradients, nerve excitation and conduction, and the regulation of the central nervous system. The important role of nanochannels has attracted people's attention, and many studies have focused on this, and the in vitro artificial nanopore technology that simulates nanochannels in living organisms h...

AI Technical Summary

Problems solved by technology

Although the ionic current signal can reflect the information of the analyte passing through the nanopore to a certain extent, the current change signal of the nanopore measured by applying a constant potential is single, which cannot directly reflect the conformational change of the analyte during the perforation process and its relationship with the nanopore. Interactions and many other information

Not only that, the large working voltage range of the constant potential method will have an impact on the stability of the nanopore

There are unavoidable technical limitations of the nanopore measurement method in the potentiostatic mode

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