Methods for Manipulating DNA Molecules in a Micrometer Scale using Optical Techniques

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Methods for Manipulating DNA Molecules in a
Micrometer Scale using Optical Techniques

• July 13, 2004
• Summarized by Ji-Yoon Park

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Abstract

• Optical method
• Control the position reaction of DNA molecules
  in a micrometer scale
• Two single laser beams for optical manipulation
  temperature control
• Independent control

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Introduction

• Optical computing
• Using inherent property of light
• Fast propagation, parallelism, and a large
  bandwidth
• Example
• Vertical cavity surface-emitting lasers (VCSELs)
  
• Diffractive optical elements (DOEs)
• DNA Light high-performance information system
• In this paper
• Methods for controlling the position reaction
  of DNA in micrometer scale

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Optically Assisted DNA Computing
Figure 1. The basic concept of optically assisted
DNA computing
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Manipulation of DNA

• Optical manipulation
• A method for non-invasively manipulating an
  object with a radiation pressure force induced
  between light and the object
• DNA cluster
• DNA strands connected to a microscopic bead

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Manipulation of DNA using Optical Techniques
Figure 2. A method for controlling the
temperature of a solution in a
micrometer scale
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Experimental Section (1/2)

• Three step operation
• Attaching the DNA cluster to the bead
• Translation of the bead
• Detaching the DNA cluster from the bead

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Experimental Section (2/2)

• Anti-tag sequence immobilization to bead
  (diameter 6 µm) 5-biotin-CATAG TACAA ATCTT
  ACCTC ATTTC AGTTA CTGAT CCACG-3
• Alexa Fluor 647 attach the tag sequence
• 5-Alexa 647- CGTGG ATCAG TAACT GAAAT
  GAGGT AAGAT TTGTACTATG-3
• Hybridization with anti-tag sequence and tag
  sequence
• Extraction of DNA cluster
• Substrate coating with titanylphthalocyanine
  (0.15 µm) gold deposition
• Thiol-modified anti-tag sequence binding to
  substrate

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Experimental Setup
Figure 3. Experimental setup
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Figure 4. Experimental result of detachment of
information DNA from a substrate
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Dependence of the Fluorescence Intensity
Figure 5. Dependence of the fluorescent intensity
on irradiation cycle when the laser beam
for temperature control is used.
(i) fluorescent molecules are attached
to tag DNA (ii) Bead on the anti-tag
DNA with fluorescent molecules
Irradiation cycle 5 mW for 15 sec stop for 4
sec
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Fluorescence Images
Figure 6. Experimental result of translation and
detachment of DNA. (a) at initial
state (b) after translation
(c) after detachment 3 mW for 1 min
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Fluorescence Intensity of Microwell Array

• anti-tag DNA attachment
• Tag DNA binding
• Irradiation (4 mW for 1 min)
• Fluorescence intensity

Figure 7. Fluorescent intensity of a DNA cluster
(a) before and (b) after irradiating
with a laser beam
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Conclusions

• Develop methods for manipulating DNA molecules in
  a micrometer scale with optical techniques
• The position and reaction of DNA are controlled
  independently using two laser beams with
  different wavelengths