2006 Fiscal Year Final Research Report Summary
Two-Color Picosecond Time-resolved Infrared Super-resolution Microscopy and its Application to Cell
| Project/Area Number |
16072207
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Science and Engineering
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
FUJII Masaaki Tokyo Institute of Technology, Chemical Resources Laboratory, Professor (60181319)
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| Co-Investigator(Kenkyū-buntansha) |
SAKAI Makoto Tokyo Institute of Technology, Chemical Resources Laboratory, Associate Professor (60298172)
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| Project Period (FY) |
2004 – 2006
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| Keywords | Two-Color / Picosecond / Infrared Super-resolution / Fluorescence / Microscope / Cell / Vibrational Relaxation |
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| Research Abstract |
The IR microscopy based on the transient fluorescence detected IR spectroscopy is capable of breaking the diffraction limit. Briefly, a tunable IR light is co-linearly introduced with a visible light of which the wavelength is fixed to slightly longer than the visible absorption band. If the frequency of the IR light does not resonant to the vibrational level, no fluorescence will appear because the visible light does not match the absorption band. When the IR frequency is resonant to the vibrational level, the vibrationally excited molecule generated by the IR excitation can absorb the visible light, and gives the transient fluorescence. Here, the transient fluorescence due to IR excitation appears only the spatial region where both IR and visible lights are overlapped. This overlapped region can be smaller than the diffraction limit of IR light and its size is improved to the diffraction limit of visible light.
In this work, we succeeded in observation of the transient fluorescence image that is smaller than the diffraction limit in rhodamine 6G/chloroform-d1 solution. We also reported the application of an IR super-resolution microscopy to a rhodamine-6G doped fluorescence bead as a microscopic target. The transient fluorescence image of a 15-μm-diameter bead is clearly observed, and the observed spatial resolution of TFD-IR image is 6.8 μm FWHM. Since the theoretical diffraction limit for the IR light is about 16.6 μm FWHM, it is clear that the spatial resolution of TFD-IR image is smaller than the IR diffraction limit, and the super-resolution is achieved for IR microscopy. In addition, the spatial resolution is determined only by visible light, not IR light. By using this infrared super-resolution microscopy, we visualized the structure and vibrational relaxation dynamics in Arabidopsis root cells.
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