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Unicell detector based on nano fiber probe and its probe manufacturing method

A nano-fiber probe and nano-probe technology, which is applied in the direction of nanotechnology for sensing, nanotechnology, nanotechnology, etc., can solve the problem that real-time monitoring cannot be realized, the size of a single cell is small, and it is not enough to find the difference at the level of a single cell issues such as improved spatiotemporal resolution and detectable target range, high sensitivity, and reduced cell sample volume

Active Publication Date: 2014-11-05
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the size of a single cell is very small, and the chemical components contained in it are also ultra-trace. However, the current analysis method Western blot needs to break millions of cells to achieve the detection of intracellular biochemical indicators. Therefore, traditional analysis Method is insufficient to detect differences at the level of individual cells
Not only that, the traditional methods are mostly fixed-point detection, which cannot realize real-time monitoring

Method used

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  • Unicell detector based on nano fiber probe and its probe manufacturing method
  • Unicell detector based on nano fiber probe and its probe manufacturing method
  • Unicell detector based on nano fiber probe and its probe manufacturing method

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Embodiment 1

[0039] The present invention develops a lactate dehydrogenase-based fiber optic nanosensor to realize lactate analysis of single cancer cells. Firstly, a polymer brush is modified on the nanoprobe to increase the immobilization amount on the nano-tip of the lactate dehydrogenase nanofiber sensor. The fluorescent by-product NADH produced by the dehydrogenase while promoting the conversion of lactate can be used for Determine the lactic acid concentration. The nanofiber sensor can achieve high spatial resolution, low background interference, and wide dynamic range (0.06-3mM) suitable for the physiological range of single-cell lactate secretion. The sensor successfully detected the diffusion curve of extracellular lactate and could clearly distinguish the lactate secretion levels of different cells such as two breast cancer cells, as Figure 4 as shown, Figure 4 The left figure in the center is the standard curve for lactate detection of nanoprobes modified with polymer brushe...

Embodiment 2

[0041] The single-cell detector of the present invention can not only be used for the research of animal cells, but also suitable for the single-cell detection of fungi and bacteria due to its nano-level probes. different subcellular locations. like Figure 5 As shown, at different hyphal sites, the measured fluorescence intensities are quite different, spanning several orders of magnitude, Figure 5 For the detection of fluorescence at the subcellular level in a single fungal cell, the fluorescence intensity of the subcellular sites (1-6) in the hyphae varies greatly. This preliminary result suggests that the fluorescent signals at subcellular sites are most likely related to key sites of neolamina metabolism. More in-depth single-cell analysis has great potential in revealing key sites and key genes in mycotoxin synthesis in cells, single-cell heterogeneity of enzymes, and the mechanism of mycotoxin cell-to-cell spread infection.

Embodiment 3

[0043] The probe combines nanoring electrodes and fiber optic sensors to simultaneously detect electrical and optical signals with high spatial resolution in real time. Figure 6 It is used for synchronous photoelectric detection of calcium ion level changes and dopamine release in PC12 cells. (a) Typical current response derived from dopamine release and (c) calcium ion fluorescence signal; (b, d) are the corresponding statistical analysis results (N = 10). a: A 24 hours after NGF withdrawal, b: Differentiation in culture medium containing 100ng / mL NGF for 3 days, c: Cells after drug withdrawal were incubated in culture medium supplemented with 150μM L-DOPA for 60 minutes . Arrows indicate the time points of high potassium solution addition. Error bars represent standard deviation; ** and * indicate p Figure 6 As shown in a and c, in the degenerated PC12 cells, not only the calcium influx was significantly reduced, but also the dopamine secretion was much reduced compared ...

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Abstract

The invention relates to an unicell detector based on a nano fiber probe, which comprises a nano probe, a light source unit, a micro operation system, an electricity detection unit, a cellular localization system and a photon detection unit, an innermost layer of the nano probe is a fiber layer, the outer wall of the fiber layer is wrapped with a nano ring electrode layer, and the outer wall of the nano ring electrode layer is wrapped with an insulating layer. A manufacturing method of the unicell detector based on the nano fiber probe comprises the steps of drawing, sputtering, making the insulating layer and cutting focused ion beam. The unicell detector has high sensitivity and can realize the unicell grade detection, compared with a traditional detection means by crushing millions of cells, the required cell sample amount is greatly reduced, and the success rate of disease detection at early stage can be increased. The unicell detector can perform in-vivo cell detection and can avoid the false appearance generation during a cell processing process. The space-time resolution and detectable target object scope can be greatly enhanced, and a biochemistry mechanism in the unicell enables real-time detection and analysis.

Description

technical field [0001] The invention relates to the application of electrical and optical technologies in the biological field, in particular to a single-cell detector and a method for making probes thereof. Background technique [0002] In recent years, with the gradual deepening of research on cell physiology, scientists have discovered that even with the same genetic material and the same surrounding environment, these cells will still develop in different directions, and sometimes produce different functions. Individual cells appear to be the same, but their chemical composition and stress response are very different, and changes in individual cells are also related to major diseases such as cancer and neurological diseases. Single-celled biological processes in plants and animals are also far from fully understood. Single-cell analysis is therefore an important means of understanding and deciphering many critical life processes. However, the size of a single cell is v...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/14B82Y15/00
Inventor 李长明郑昕婷
Owner SOUTHWEST UNIV
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