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Methods and Devices for Charged Molecule Manipulation

a charge molecule and charge technology, applied in the field of micromanipulation of charged molecules, can solve the problem of often problematic microinjection of foreign materials

Inactive Publication Date: 2010-12-23
BRIGHAM YOUNG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0020]As used herein, the term “about” is used to provide flexibility to a numerical range endpoint by providing that a given value may be “a little above” or “a little below” the endpoint without affecting the desired result.

Problems solved by technology

Microinjection of foreign materials is often problematic, particularly if such microinjection is being performed on a biological structure such as a living cell.

Method used

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  • Methods and Devices for Charged Molecule Manipulation
  • Methods and Devices for Charged Molecule Manipulation
  • Methods and Devices for Charged Molecule Manipulation

Examples

Experimental program
Comparison scheme
Effect test

example 1

DNA Visualization and Imaging Methods

[0053]4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) is used to visualize the DNA in the following example. DAPI exhibits low toxicity and its strong fluorescence under ultra-violet illumination. When dissolved in water and not bound to DNA, DAPI has an excitation maximum of 355 nm (ultra-violet light) and an emission maximum of 453 nm (blue light). When DAPI is bound to DNA its excitation maximum changes to 388 nm and its emission maximum shifts to 454 nm, and the intensity of the emitted light increases roughly twenty-fold compared to free DAPI. The increase in emission intensity when DAPI binds with DNA makes it possible to distinguish between unbound DAPI and DAPI-stained DNA.

[0054]DAPI-stained DNA is visualized using a Zeiss Axioskop Fluorescence Microscope with UV illumination and a purpose-built blue light filter for imaging DAPI stained samples. Because the DAN-DNA complex fluoresces in the blue portion of the visible spectrum, the ...

example 2

DNA Attraction Experiment

[0055]The DNA attraction and repulsion experiments are performed both in distilled water and in 0.9% saline solution. In both cases, the experiments follow identical protocols, with

the exception of the media into which the device is submerged. A MEMS device as has been described herein is covered in approximately 2 mm of either distilled water or 0.9% saline solution. A 1-2 μL drop of 306 ng / μL DAPI stained DNA is placed in the solution near the device using a calibrated pipette. The needle structure of the MEMS device is connected to the positive terminal of a voltage source providing 1.5 V DC. The substrate of the MEMS device is connected to the negative terminal of the voltage source. Images can be taken to verify the DAPI-stained DNA on the surface of the needle structure. Approximate concentrations of the DNA can be calculated using the linear model of Equation I.

example 3

DNA Repulsion Experiment

[0056]DNA is attracted to the tip of a MEMS needle structure as is described in Example 2, by connecting the needle structure to the positive terminal of a 1.5 V DC voltage source and connecting the MEMS device substrate to the negative terminal of the voltage source. Following attraction of DNA, the polarity of the electrical charge is then reversed so that the positive terminal is connected to the MEMS device substrate and the negative terminal is connected to the needle structure. Images can be taken from the time the polarity is reversed to verify DNA release and repulsion. Additionally, the time between connecting the MEMS needle structure to the negative terminal and when DNA is clearly repelled from the needle structure can be calculated, and approximate concentrations can be calculated using the linear model given in Equation I.

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Abstract

Systems and methods for manipulating molecular material are provided. In one aspect, for example, a method for manipulating molecular material may include positioning an uncharged needle structure in electrical proximity with a charged molecular material at a first locus in a liquid environment, charging the needle structure such that at least a portion of the charged molecular material becomes associated with the needle structure, moving the needle structure and the first locus relative to one another, and discharging the needle structure to disassociate the charged molecular material at a second locus.

Description

FIELD OF THE INVENTION[0001]The present invention relates to the micromanipulation of charged molecules. Accordingly, this invention involves the fields of biotechnology, chemistry, and micromanipulation.BACKGROUND OF THE INVENTION[0002]Microinjection of foreign materials is often problematic, particularly if such microinjection is being performed on a biological structure such as a living cell. Various transfection techniques include the microinjection of foreign genetic material such as DNA into the nucleus of a cell to facilitate the expression of foreign DNA. For example, when a fertilized oocyte (egg) is transfected, cells arising from that oocyte will carry the foreign genetic material. Thus in one application organisms can be produced that exhibit additional, enhanced, or repressed genetic traits. As one example, researchers have used microinjections to create strains of mice that carry a foreign genetic construct causing macrophages to auto-fluoresce and undergo cell death w...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N13/00C12M1/42
CPCC12N15/89C12M35/02G01N33/48
Inventor ATEN, QUENTIN T.HOWELL, LARRY L.JENSEN, BRIAN D.BURNETT, SANDRA
Owner BRIGHAM YOUNG UNIV