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Spectrally-resolved fluorescence lifetime imaging microscope using tunable bandpass filters

Spectrally-resolved fluorescence lifetime imaging microscope using tunable bandpass filters. Hyeong -Jun Jeong , Jin-Woo Ahn , Dukho Do and Dae -Gab Gweon. 2.8 ns. C.W: 490nm. 500nm. 510nm. 2.0 ns.

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Spectrally-resolved fluorescence lifetime imaging microscope using tunable bandpass filters

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  1. Spectrally-resolved fluorescence lifetime imaging microscope using tunable bandpass filters Hyeong-Jun Jeong, Jin-Woo Ahn, DukhoDo and Dae-Gab Gweon 2.8 ns C.W: 490nm 500nm 510nm 2.0 ns A simple structure of spectral fluorescence lifetime imaging microscope (SLIM) isdesigned with the use of tunable bandpass filter, a kind of Fabry-perot filter that transmission wavelength is varying according to incident angle of light. Using two angle-tuned bandpass filter(ATBF) in series, spectral bandwidth can be adjustable down to 4 nm. We obtained spectrally resolved FLIM images and confirmed that it provides high accuracy to lifetime distribution and high contrast images. 520nm 530nm 540nm

  2. Pulse laser (780 nm, 80MHz, 100fs, 140 mW) Stop <Spectral bandwidth adjustment with the use of dual ATBFs> <Schematic diagram of Spectral FLIM system> Angle-tuned bandpass filter Collecting lens Wedeveloped a simple structure of spectral fluorescence lifetime imaging microscope (SLIM) with the use of tunable bandpass filter, a kind of Fabry-perot filter that transmission wavelength is varying according to incident angle of light. Feasibility tests of this angle-tuned bandpass filter (ATBF) are performed and it shows high transmission and constant spectral bandwidth (20 nm) with respect to angle of incidence (AOI). Furthermore, using two ATBFs in series, spectral bandwidth can be adjustable down to 4 nm. In this paper, dual ATBFs are implemented to the detection part of FLIM system so that we obtained spectrally resolved FLIM images. We compare these SLIM images with an original FLIM image and confirm that the former case provides high accuracy to analyze lifetime distribution as well as high contrast of images. The proposed SLIM microscope with good wavelength selectivity has many opportunities to utilize to other applications such as FLIM-FRET and autofluorescence imaging. This paper was accepted by the journal of Review of Scientific Instrument (RSI), Volume 83 (9), on August 25, 2012. Start Dichroic mirror PMT TCSPC Data storage & processing Objective Sample X,Y scanner Relay optics Pixel, line, frame sync

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