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Recyclable hollow cantilever probe front end loading method and cleaning method

A technology of cantilever beam probes and probes, which is applied in the direction of liquid cleaning methods, cleaning methods and utensils, chemical instruments and methods, etc., can solve the problems of low liquid utilization rate and cumbersome cleaning steps, and reduce the cleaning workload , improve the utilization rate and reduce the effect of exchange time

Active Publication Date: 2022-02-25
SOUTH CHINA UNIV OF TECH
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Problems solved by technology

[0005] Aiming at the problems of low liquid utilization rate and cumbersome cleaning steps of the above-mentioned FluidFM hollow cantilever probe in the application scenario of high operating liquid cost and low consumption, the present invention provides a A recyclable method for loading and cleaning the front end of a hollow cantilever probe. By loading t

Method used

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  • Recyclable hollow cantilever probe front end loading method and cleaning method
  • Recyclable hollow cantilever probe front end loading method and cleaning method
  • Recyclable hollow cantilever probe front end loading method and cleaning method

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Embodiment 1

[0028] Such as figure 1 As shown, the present embodiment provides a recyclable method for loading and cleaning the front end of a hollow cantilever probe, comprising the following steps:

[0029] S1 disinfects and cleans the operating devices.

[0030] S2 Filter and degas the liquid to be operated; transfer the processed liquid to a transparent substrate; contact the calibrated hollow probe with the droplet; apply negative pressure pulses through the pressure controller to complete the front loading of the liquid .

[0031] S3 conducts research experiments on single-cell level manipulation.

[0032] S4 After the test is completed, the probe is immersed in the cleaning solution for a while, and cleaned with ultrapure water to complete the external cleaning; after that, the probe is immersed in sodium hypochlorite solution and ethanol aqueous solution in turn, and the loading and releasing operations are alternately performed during the period to complete the inside of the pro...

Embodiment 2

[0043] It is basically the same as Embodiment 1, except that this embodiment provides an embodiment of micro-droplet patterning after loading by the front end of the probe. Such as figure 2 As shown, the steps are as follows:

[0044] First, clean the transparent glass substrate 101, dry it with nitrogen, and place it in a clean fume hood; use a syringe filter with a pore size of 0.1 μm to filter the fluorescent droplets 102 (rose bengal 6G fluorescent dye aqueous solution), and pass The micropipette transfers about 50 μL of the filtered fluorescent droplet 102 to the transparent glass substrate 101 .

[0045] Then place the transparent glass substrate 101 with fluorescent droplets 102 on the observation window of FluidFM, adjust the position of the transparent glass substrate 101 so that the fluorescent droplets 102 are above the objective lens 100 of the inverted fluorescent microscope, and limit and fix the transparent glass substrate 101 ; Install the hollow cantilever ...

Embodiment 3

[0053] It is basically the same as that of Example 2, except that this example provides an example of carrying out cell-targeted labeling after the front end of the probe is loaded.

[0054] According to Example 2, the front-end loading of the fluorescent labeling solution was completed.

[0055] Place the cell culture dish prepared in advance on the observation window of the inverted fluorescence microscope, and add or reduce the PBS solution to the culture dish so that the liquid level in the dish is about 3 mm, which is convenient for subsequent probe operations.

[0056] Make the probe approach the upper surface of the culture dish step by step. After the tip of the probe is immersed in the culture dish solution, adjust the laser spot so that the position of the light spot is in the center of the laser receiver. Then look for the cells to be operated from the window, and make the probe gradually approach the cells to about 100 μm after confirming the operation of the cells...

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Abstract

The invention discloses a recyclable hollow cantilever probe front end loading method and cleaning method, and solves the problems of low liquid utilization rate and tedious cleaning steps of a hollow cantilever probe in an application scene of high operating liquid cost and low using amount. The front end loading method comprises the following steps: filtering and degassing liquid to be operated; transferring the liquid drops onto a transparent substrate; contacting the calibrated probe with the liquid drop; applying negative pressure pulse through the pressure controller to complete front-end loading of the liquid. The invention also provides the cleaning method after front-end loading. The cleaning method comprises the following steps: immersing the probe subjected to the test in a cleaning solution for a moment, washing with ultrapure water to complete external cleaning, then immersing the probe in a sodium hypochlorite solution and an ethanol water solution in sequence, alternately applying positive pressure and negative pressure during the period to realize internal cleaning of the microchannel, and finally, storing the cleaned probe in a wet, sealed and refrigerated environment so as to make the hollow cantilever probe quickly and continuously used next time.

Description

technical field [0001] The invention relates to the technical field of micro-nano operation, in particular to a recyclable method for loading and cleaning the front end of a hollow cantilever probe. Background technique [0002] Physiological studies at the single-cell level are fundamental to further understanding cellular processes in complex tissues and organs. Among them, the response of cells to external stimuli can provide more details and principles, and fluid force microscopy (Fluidic Force Microscopy, FluidFM) realizes the release of local neurotransmitters and drugs to cells through force feedback and pressure-controlled hollow cantilever probes , providing a powerful tool for single-cell level research. [0003] In 2009, the Meister team combined the hollow probe and commercial AFM to create FluidFM, and rapidly expanded the field of biomedical research (Meister A, Gabi M, Behr P, et al.FluidFM: Combining AtomicForce Microscopy and Nanofluidics in a Universal Liq...

Claims

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Application Information

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IPC IPC(8): B01L3/00B08B3/04
CPCB01L3/5027B08B3/04
Inventor 张宪民杨倬波冯柯崔超宇李海
Owner SOUTH CHINA UNIV OF TECH
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