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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: 2010-09-30
BRIGHAM YOUNG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0010]In another aspect of the present invention, a cellular constraint device can include a support structure having an initial position and an extended position. The support structure is moveable from the initial position to the extended position, and when in the extended position, the support structure is operable to constrain a cell. In yet another aspect, the support structure when in the initial position is operable to receive the cell. In one specific aspect, the support structure is operable to constrain the cell by contacting the cell above a midline of the cell. In another specific aspect, the support structure is operable constrain the cell and to prevent lateral displacement of the cell.
[0022]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
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Effect test

example 1

DNA Visualization and Imaging Methods

[0055]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.

[0056]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 DAPI-DNA complex fluoresces in the blue portion of the visible spectrum, the...

example 2

DNA Attraction Experiment

[0057]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

[0058]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

Devices and methods for constraining or holding a cell are provided. In one aspect, for example, a cellular constraint device is provided. Such a device can include a support surface and at least one constraining arm movably coupled to the support surface. The constraining arm has a first position in which the constraining arm is substantially parallel and substantially adjacent to the support surface. Additionally, at least a portion of the constraining arm is movable away from the support surface to a second position where the constraining arm is operable to constrain a single cell.

Description

PRIORITY DATA[0001]This application is a continuation of U.S. patent application Ser. No. 12 / 668,369, filed on Jan. 8, 2010, which is a U.S. nationalization of PCT Application No. PCT / U.S.08 / 69550 filed on Jul. 9, 2008, which claims the benefit of U.S. Provisional Patent Application Ser. No. 60 / 958,624, filed on Jul. 9, 2007, all of which are incorporated herein by reference in their entireties.GOVERNMENT INTEREST[0002]This invention was made with government support under National Science Foundation Grant No. 0428532. The United States government has certain rights to this invention.FIELD OF THE INVENTION[0003]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[0004]Microinjection of foreign materials is often problematic, particularly if such microinjection is being performed on a biological structure such as a living cell. Various ...

Claims

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Application Information

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