On-chip single-cell-based gene expression analysis using gold nano-particles

Hyonchol Kim, Atsushi Kira, Hironobu Kohno, Kazunori Matsumura, Kazuki Orita, Koudai Oikawa, Tomoyuki Kaneko, Kazunori Okano, Kenji Yasuda

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

The properties of individual cells are thought to depend on the time course change of the number and the spatial distribution of regulatory molecules in cells. To understand the strict meaning of those roles of some particular sets of regulatory proteins, we need to measure both the phenotypical characteristics of cells and the quantitative amount of mRNAs in each of those individual cells. For that purpose, we have developed a novel method to measure the quantitative amount of mRNA expression in individual cells keeping their spatial distributions in the cell without any amplification process like PCR. In this method, a set of different sizes of gold nano-particles attached with different probe-DNA respectively were used as a set of probes to detect different mRNAs existing in a cell. At first, the optimum condition of the immobilization of probe-DNA onto the gold nano-particle surface was examined. Next, the selectivity of the probe-DNA immobilized onto the nano-particle was tested using complementary oligonucleotide molecules. We confirmed the several different kinds of gold nano-particle probes were hybridized with target oligonucleotides having complementary sequences with almost 100% selectively. For the counting and distinguishing each of the gold nano-particles, we used two different methods and compared: one is Atomic Force Microscopy and the other is Scanning Electron Microscopy. Quantitative detection of mRNAs in individual cells keeping their spatial distributions was then examined using the gold nano-particle-based detection system. In this meeting, we will present the results and will discuss about the potential and problems of this method for the single-cell-based quantitative expression analysis.

Original languageEnglish
Title of host publicationDigest of Papers - Microprocesses and Nanotechnology 2005: 2005 International Microprocesses and Nanotechnology Conference
Pages44
Number of pages1
Volume2005
Publication statusPublished - 2005
Externally publishedYes
Event2005 International Microprocesses and Nanotechnology Conference - Tokyo
Duration: 2005 Oct 252005 Oct 28

Other

Other2005 International Microprocesses and Nanotechnology Conference
CityTokyo
Period05/10/2505/10/28

Fingerprint

Gene expression
Gold
Spatial distribution
DNA
Oligonucleotides
Molecules
Amplification
Atomic force microscopy
Proteins
Scanning electron microscopy
Messenger RNA

ASJC Scopus subject areas

  • Engineering(all)

Cite this

Kim, H., Kira, A., Kohno, H., Matsumura, K., Orita, K., Oikawa, K., ... Yasuda, K. (2005). On-chip single-cell-based gene expression analysis using gold nano-particles. In Digest of Papers - Microprocesses and Nanotechnology 2005: 2005 International Microprocesses and Nanotechnology Conference (Vol. 2005, pp. 44). [1595205]

On-chip single-cell-based gene expression analysis using gold nano-particles. / Kim, Hyonchol; Kira, Atsushi; Kohno, Hironobu; Matsumura, Kazunori; Orita, Kazuki; Oikawa, Koudai; Kaneko, Tomoyuki; Okano, Kazunori; Yasuda, Kenji.

Digest of Papers - Microprocesses and Nanotechnology 2005: 2005 International Microprocesses and Nanotechnology Conference. Vol. 2005 2005. p. 44 1595205.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Kim, H, Kira, A, Kohno, H, Matsumura, K, Orita, K, Oikawa, K, Kaneko, T, Okano, K & Yasuda, K 2005, On-chip single-cell-based gene expression analysis using gold nano-particles. in Digest of Papers - Microprocesses and Nanotechnology 2005: 2005 International Microprocesses and Nanotechnology Conference. vol. 2005, 1595205, pp. 44, 2005 International Microprocesses and Nanotechnology Conference, Tokyo, 05/10/25.
Kim H, Kira A, Kohno H, Matsumura K, Orita K, Oikawa K et al. On-chip single-cell-based gene expression analysis using gold nano-particles. In Digest of Papers - Microprocesses and Nanotechnology 2005: 2005 International Microprocesses and Nanotechnology Conference. Vol. 2005. 2005. p. 44. 1595205
Kim, Hyonchol ; Kira, Atsushi ; Kohno, Hironobu ; Matsumura, Kazunori ; Orita, Kazuki ; Oikawa, Koudai ; Kaneko, Tomoyuki ; Okano, Kazunori ; Yasuda, Kenji. / On-chip single-cell-based gene expression analysis using gold nano-particles. Digest of Papers - Microprocesses and Nanotechnology 2005: 2005 International Microprocesses and Nanotechnology Conference. Vol. 2005 2005. pp. 44
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