Highly-sensitive fluorescence detection and imaging with microfabricated total internal reflection (TIR)-based devices

Nam Cao Hoai Le, Dzung Viet Dao, Ryuji Yokokawa, Thien Duy Nguyen, John C. Wells, Susumu Sugiyama

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

This paper presents the development and application of several total internal reflection (TIR)-based devices for highly-sensitive and high-resolution fluorescence imaging. Using micro electro mechanical systems (MEMS) fabrication technology, miniaturized single-wavelength and dual-wavelength TIR-based devices have been designed and fabricated. A low-cost and simple fabrication process utilizing TMAH wet etching, deep reactive ion etching (DRIE) and polymer casting has enabled us to integrate several optical components into one single poly(dimethylsiloxane) (PDMS) chip, thus alignment and assembly are eliminated. The slide-format and monolithic chip can be used with both upright and inverted fluorescent microscope with interchangeable sample delivery platforms, i. e. glass slides, flow-cells, microchannels etc... In different configurations, the devices were applied in measurement of the average velocity field of fluorescent beads in pressure-driven flow in a microchannel and then in detecting of single DNA molecules. Another ongoing effort is focused on using the device for cell imaging which will be briefly discussed. These applications are the first step toward integration the TIR-based chips into a detection functionality of a μ-TAS. Our proposed devices are smaller, simpler and less expensive comparing to current objective-type and prism-type total internal reflection fluorescent microscopy (TIRFM) systems. At this moment, our devices could provide quick and low-cost evanescent excitations with one or two wavelengths in upright or inverted fluorescent microscopes using normal objective lenses. In the future, they might be integrated as a detection module in a μ-TAS where highly-sensitivity and high-resolution fluorescence imaging capability is necessary. Finally, some discussion on the limitation and future developments for our devices will be also provided.

Original languageEnglish
Pages (from-to)45-59
Number of pages15
JournalJournal of Micro-Nano Mechatronics
Volume7
Issue number1-3
DOIs
StatePublished - Dec 2012
Externally publishedYes

Funding

Acknowledgment N. C. H. Le gratefully acknowledges the Ministry of Education, Culture, Sports, Science and Technology, Government of Japan (MEXT) of Japan for his doctoral scholarship. This study was partially supported by the Kyoto NanoTech Cluster and Research for Promoting Technological Seeds grant from Japan Science and Technology Agency (JST). The authors thank Prof. M. Kasahara with Department of Bioscience and Biotechnology, Ritsumeikan University for helping the preparation of DNA fragments.

FundersFunder number
Kyoto NanoTech Cluster and Research for Promoting Technological Seeds
Ministry of Education, Culture, Sports, Science and Technology, Government of Japan
Ministry of Education, Culture, Sports, Science and Technology
Japan Science and Technology Agency

    Keywords

    • Cell imaging
    • Fluorescent imaging
    • Micro electro mechanical systems (MEMS)
    • Micro-total analysis systems (micro-TAS)
    • Nano-particle image velocimetry (nano-PIV)
    • Single molecule detection
    • Total internal reflection (TIR)-based chip

    Fingerprint

    Dive into the research topics of 'Highly-sensitive fluorescence detection and imaging with microfabricated total internal reflection (TIR)-based devices'. Together they form a unique fingerprint.

    Cite this