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 single cells are small in size, cannot realize real-time monitoring, and are not enough to detect differences at the level of individual cells problems such as high sensitivity, improved spatiotemporal resolution and detectable target range, and the effect of reducing the amount of cell samples

Active Publication Date: 2013-05-15
SOUTHWEST UNIVERSITY
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  • Abstract
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  • 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 an optical fiber nanosensor based on lactate dehydrogenase to realize lactate analysis of a single cancer cell. First modify a layer of polymer brush on the nanoprobe to increase the amount of fixation on the nanotip of the lactate dehydrogenase nanofiber sensor. This dehydrogenase promotes the conversion of lactic acid and produces the fluorescent by-product NADH, which can be used for Determine the lactic acid concentration. The nano fiber sensor can achieve high spatial resolution, low background interference, and a wide dynamic range (0.06-3mM) suitable for the physiological range of single cell lactate secretion. The sensor successfully detects the diffusion curve of extracellular lactic acid, and can clearly distinguish the lactic acid secretion levels of different cells such as two breast cancer cells, such as Figure 4 As shown, Figure 4 The middle left picture shows the standard curve of lactic acid detection of the nanoprobe modi...

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 single-cell detection of fungi and bacteria due to its nanometer-level probes. It only needs to precisely pierce the nanometer probe into the fungal cell hyphae. Different subcellular sites. Such as Figure 5 As shown, at different mycelial sites, the measured fluorescence intensity is very different, spanning several orders of magnitude, Figure 5 It is a subcellular level fluorescence detection in a single fungal cell. The fluorescence intensity of subcellular sites (1-6) in mycelium varies greatly. This preliminary result indicates that the fluorescence signal at the subcellular site is very likely to be related to the key part of the new layer of metabolism. More in-depth single-cell analysis has great potential in revealing the key sites and genes in the synthesis of mycotoxins in cells, the single-cell heterogeneity of enzymes, and the mech...

Embodiment 3

[0043] The probe combines a nano-ring electrode and an optical fiber sensor, which can simultaneously detect electrical and optical signals with high spatial resolution in real time. Image 6 For synchronous photodetection 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 : 24 hours after stopping nerve growth factor A, b : Differentiation for 3 days in culture liquid containing 100ng / mL NGF, c : After drug withdrawal, the cells were incubated for 60 minutes in a medium supplemented with 150μM L-DOPA. The arrow indicates the time point when the high potassium solution was added. Error bars represent standard deviation; ** and * represent p Image 6 As shown in a and c, in the degenerated PC12 cells, not only the calcium influx is significantly reduced, but also the dopamine secretion is much r...

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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 Applications(China)
IPC IPC(8): G01N15/14B82Y15/00
Inventor 李长明郑昕婷
Owner SOUTHWEST UNIVERSITY
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