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Method and apparatus for the rapid disruption of cells or viruses using micro beads and laser

a technology of applied in the field of rapid disruption of cells or viruses using micro beads and lasers, can solve the problems of harsh chemicals used to disrupt cells, labor-intensive and time-consuming chemical methods, and chemical lysis methods, and achieve the effects of efficient amplification, efficient amplification, and increased pcr efficiency

Inactive Publication Date: 2008-08-21
SAMSUNG ELECTRONICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a method and apparatus for disrupting cells or viruses using magnetic beads and a laser. The method involves mixing beads with a solution containing cells or viruses, vibrating the beads, and irradiating the beads to heat and disrupt the cells or viruses. The apparatus includes a member for receiving a cell lysis chamber, a vibrator for mixing the sample and magnetic beads, a laser generator for supplying a laser, and an anti-evaporation part to prevent the sample from evaporating. The cell lysis chip has a reaction chamber, an inlet hole, and an outlet hole, and is designed to allow a laser to pass through and has an inlet hole and an outlet hole. The chip bottom is attached to the chip body through a chip bonding part to close the lower portion of the reaction chamber, the inlet hole, and the outlet hole. The technical effects of this invention include improved efficiency and accuracy in cell or virus disruption, reduced risk of sample evaporation, and improved safety during the process.

Problems solved by technology

The chemical lysis method is disadvantageous in that harsh chemicals are used to disrupt the cells.
Since they can interfere with the subsequent PCR, it is necessary to purify the DNA prior to the PCR.
The chemical method is labor-intensive and time-consuming, requires expensive consumables and has often a low DNA yield.
Simple heating is disadvantageous in that it results in the denaturation of proteins, which can be attached to released DNA.
They can also interfere with DNA amplification.
A physical method uses a bulky and expensive pressure apparatus, which is not suitable for a Lab-on-a-Chip (LOC) application.
First, a distribution of ultrasonic energy is not uniform.
The nonuniform distribution of ultrasonic energy leads to inconsistent results.
Second, due to the energy divergence in the ultrasonic bath, it takes often several minutes to completely disrupt cells.
Lastly, the ultrasonic method generates unpleasant sounds.
This method causes only a loss of function of the cells by using a laser and nanoparticles and there is no description of a method of disrupting cells by vibrating a solution containing cells and particles.

Method used

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  • Method and apparatus for the rapid disruption of cells or viruses using micro beads and laser
  • Method and apparatus for the rapid disruption of cells or viruses using micro beads and laser
  • Method and apparatus for the rapid disruption of cells or viruses using micro beads and laser

Examples

Experimental program
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Effect test

preparation example 1

Cell Lysis System

[0151]As illustrated in FIG. 1, bacterial cells prepared as describe below (90 μl) and micro magnetic beads (30 μl, Dynabeads® M-270 Carboxylic Acid, DYNAL, Norway) were mixed in a vial located in a vial guide (AMITECH, Korea). 808 nm, 13.8 W high power laser beam (HLU25F100-808, LIMO, Germany) was applied to the mixture for disrupting cells for a specific time in individual experiments while stirring the vial by vortexing (see FIG. 1).

preparation example 2

Bacterial Strain and the Determination of Bacterial Cell Viability

[0152]E. coli strain BL21 and Streptococcus mutans (ATCC# 35668) were cultured at 37° C. with vigorous aeration in brain heart infusion (BHI) media to exponential phase (OD600=0.5˜1.0). The bacterial cells were harvested by centrifugation and washed twice with 3 ml of phosphate-buffered saline (PBS) solution. The cells were resuspended in PBS (cell density; 1×105 cells / μl). The number of viable cells was determined by the ability of single cells to form colonies. Aliquots of E. coli cells (1×103) after laser beam radiation were spread onto BHI plates. The plates were incubated at 37° C. overnight, and the number of colonies was counted.

[0153]A Staphylococcus epidermidis (ATCC#14990→12228) was cultured at 37° C. with vigorous aeration in Nutrient Agar (NA) media to exponential phase (OD600=0.5˜1.0). The bacterial cells were harvested by centrifugation and washed twice with 3 ml of phosphate-buffered saline (PBS) soluti...

preparation example 3

Extraction of Bacterial Genomic DNA

[0154]In order to compare the efficiency of DNA release by laser method with the efficiency of other known conventional methods, E. coli genomic DNA (from 0.9×105 cells equivalent to the number of cells used for each laser lysis) was prepared using the boiling method for 5 min at 95° C.

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Abstract

A method and apparatus for rapid disruption of cells or viruses using beads and a laser are provided. According to the method and apparatus for rapid disruption of cells or viruses using beads and a laser, cell lysis within 40 seconds is possible, the apparatus can be miniaturized using a laser diode, a DNA purification step can be directly performed after a disruption of cells or viruses, and a solution containing DNA can be transferred to a subsequent step after cell debris and beads to which inhibitors of a subsequent reaction are attached are removed with an electromagnet. In addition, by means of the cell lysis chip, an evaporation problem is solved, vibrations can be efficiently transferred to cells through magnetic beads, a microfluidics problem on a rough surface is solved by hydrophobically treating the inner surface of the chip, and the cell lysis chip can be applied to LOC.

Description

CROSS-REFERENCE TO RELATED PATENT APPLICATIONS[0001]This application is a continuation-in-part of U.S. application Ser. No. 11 / 253,541, filed Oct. 19, 2005, which claims priority to Korean Patent Application No. 10-2004-0083586, filed on Oct. 19, 2004, Korean Patent Application No. 10-2005-0038988, filed on May 10, 2005, and Korean Patent Application No. 10-2005-0078886, filed on Aug. 26, 2005, in the Korean Intellectual Property Office, the disclosures of which are incorporated herein in their entirety by reference.BACKGROUND OF THE INVENTION[0002]1. Field of the Invention[0003]The present invention relates to a method and apparatus for rapid disruption of cells or viruses using micro beads and a laser.[0004]2. Description of the Related Art[0005]Generally, a molecular diagnosis of a specific pathogenic bacteria is performed in four steps: 1) cell lysis, 2) DNA isolation, 3) DNA amplification and 4) DNA detection.[0006]An efficient extraction of DNA from a cell is required in many ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N13/00C12M1/00
CPCB01F11/0266B01F13/0059B01L3/502761C12N13/00C12M47/06C12N1/066B01L2300/0816B01F31/86B01F33/30
Inventor LEE, JEONG-GUNKWON, YOUNG-NAMKIM, YOUNG-ALEE, MYO-YONGYOO, SHIN-ICHO, YEON-JACHEONG, KWANG-HOYOO, CHANG-EUNYANG, SEUNG-YEON
Owner SAMSUNG ELECTRONICS CO LTD
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