Single-protein molecular interactions on polymer-modified glass substrates for nanoarray chip application using dual-color TIRFM

Daekwang Kim, Gu Lee Hee, Hyungil Jung, Ho Kang Seong

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The immobilization of proteins and their molecular interactions on various polymer-modified glass substrates [i.e. 3-aminopropyltriethoxysilane (APTS), 3-glycidoxypropyltrimethoxysilane (GPTS), poly (ethylene glycol) diacrylate (PEG-DA), chitosan (CHI), glutaraldehyde (GA), 3-(trichlorosilyl)propyl methacrylate (TPM), 3'-mercaptopropyltrimethoxysilane (MPTMS), glycidyl methacrylate (GMA) and poly-l-lysine (PL).] for potential applications in a nanoarray protein chip at the single-molecule level was evaluated using prism-type dual-color total internal reflection fluorescence microscopy (dual-color TIRFM). A dual-color TIRF microscope, which contained two individual laser beams and a single high-sensitivity camera, was used for the rapid and simultaneous dual-color detection of the interactions and colocalization of different proteins labeled with different fluorescent dyes such as Alexa Fluor® 488, Qdot® 525 and Alexa Fluor® 633. Most of the polymer-modified glass substrates showed good stability and a relative high signal-to-noise (S/N) ratio over a 40-day period after making the substrates. The GPTS/CHI/GA-modified glass substrate showed a 13.5-56.3% higher relative S/N ratio than the other substrates. 1% Top-Block in 10 mM phosphate buffered saline (pH 7.4) showed a 99.2% increase in the blocking effect of non-specific adsorption. These results show that dual-color TIRFM is a powerful methodology for detecting proteins at the single-molecule level with potential applications in nanoarray chips or nano-biosensors.

Original languageEnglish
Pages (from-to)783-790
Number of pages8
JournalBulletin of the Korean Chemical Society
Volume28
Issue number5
DOIs
Publication statusPublished - 2007 May 20

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Molecular interactions
Fluorescence microscopy
Polymers
Color
Glass
Substrates
Chitosan
Glutaral
Proteins
Signal to noise ratio
Molecules
Methacrylates
Prisms
Fluorescent Dyes
Biosensors
Lysine
Laser beams
Microscopes
Cameras
Phosphates

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

Cite this

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title = "Single-protein molecular interactions on polymer-modified glass substrates for nanoarray chip application using dual-color TIRFM",
abstract = "The immobilization of proteins and their molecular interactions on various polymer-modified glass substrates [i.e. 3-aminopropyltriethoxysilane (APTS), 3-glycidoxypropyltrimethoxysilane (GPTS), poly (ethylene glycol) diacrylate (PEG-DA), chitosan (CHI), glutaraldehyde (GA), 3-(trichlorosilyl)propyl methacrylate (TPM), 3'-mercaptopropyltrimethoxysilane (MPTMS), glycidyl methacrylate (GMA) and poly-l-lysine (PL).] for potential applications in a nanoarray protein chip at the single-molecule level was evaluated using prism-type dual-color total internal reflection fluorescence microscopy (dual-color TIRFM). A dual-color TIRF microscope, which contained two individual laser beams and a single high-sensitivity camera, was used for the rapid and simultaneous dual-color detection of the interactions and colocalization of different proteins labeled with different fluorescent dyes such as Alexa Fluor{\circledR} 488, Qdot{\circledR} 525 and Alexa Fluor{\circledR} 633. Most of the polymer-modified glass substrates showed good stability and a relative high signal-to-noise (S/N) ratio over a 40-day period after making the substrates. The GPTS/CHI/GA-modified glass substrate showed a 13.5-56.3{\%} higher relative S/N ratio than the other substrates. 1{\%} Top-Block in 10 mM phosphate buffered saline (pH 7.4) showed a 99.2{\%} increase in the blocking effect of non-specific adsorption. These results show that dual-color TIRFM is a powerful methodology for detecting proteins at the single-molecule level with potential applications in nanoarray chips or nano-biosensors.",
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Single-protein molecular interactions on polymer-modified glass substrates for nanoarray chip application using dual-color TIRFM. / Kim, Daekwang; Hee, Gu Lee; Jung, Hyungil; Seong, Ho Kang.

In: Bulletin of the Korean Chemical Society, Vol. 28, No. 5, 20.05.2007, p. 783-790.

Research output: Contribution to journalArticle

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