The Claude Research GroupTitle: Microsoft PowerPoint - HCM-IFTS Author: Claude Li Created Date:...
Transcript of The Claude Research GroupTitle: Microsoft PowerPoint - HCM-IFTS Author: Claude Li Created Date:...
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Hyperspectral Microscope based on
Imaging Fourier transform Spectrometry
Dr. Li Jianping, Claude
June 2017
The Claude Research Group
Copyright: Claude
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Outline
Spectral Imaging
Fourier transform Imaging Spectroscopy
A general-purpose FTIS instrument
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• 2D-Imaging
• 1D-Spectroscopy
Spectral Imaging (Imaging Spectroscopy)
Acquisition timeTime Resolution
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Why Spectral Imaging?
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Applications of Spectral Imaging
• Astronomy
• Remote sensing
• Meteorology
• Oceanography
• Mining geology
• Mineralogy
• Agriculture
• Surveillance
• ……
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Applications in Medicine, Biology and Chemistry
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Modalities of Spectral Imaging
Imaging Spectroscopy
• Microscopy
• Photography
• Telescopy
• Reflection
• Absorption
• Transmission
• Scattering
• Raman
• Fluorescence
• Electroluminescence
• Chemoluminescence
• Bioluminescence
• ……
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Methods for Spectral Imaging
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Fourier Transform Spectrometer (FTS)
0
0
( ) ( ) ( ) ( ) cos 2I x I x I x B xd
2 exp 2B I x j x dx
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Fourier Transform Imaging Spectrometer (FTIS)
( , ) ( ) cos(2 cos ')p pI x B x d d
2 , exp 2c pB I x j x dx
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Throughput (Jacquinot) Advantage
• FTSs can have more than 60 times higher energy-
gathering capability than grating spectrometers for the
same resolving power and similar instrument size.
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Multiplex (Fellgett) Advantage
• FTS simultaneously observes all the spectral information from the
entire range of a given spectrum during a scan period.
FT
Mixed
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Wavenumber (Connes) Advantage
• The wavenumber scale of an FTIR is derived from a He-Ne laser fringe that
acts as internal references for sampling positions in each scan. Therefore,
the wavenumber calibration of FTS is much more accurate and has much
better long-term stability than the calibration of dispersive instruments.
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High and Variable Spectral Resolution
Instrumental lineshape:
11/ 2 (cm )L
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Wide Spectral Range
• Nyquist limit wavelength
• System spectral transmission response
▫ Detector spectral response
▫ Optics spectral characteristics
min 2 x
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A UV-Vis-NIR FTIS
Setup of the UV-Vis-NIR FTIS based on the beam-folding position-tracking technique
Jianping. Li, Robert. Chan, and Xuzhu. Wang, Tests of a practical visible-NIR imaging Fourier transform spectrometer for biological and chemical fluorescence emission measurements. Opt. Express, 2009. 17(23).
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Prototyping and testing (Gen1)
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Testing Summary
Mode Determinants/Typical value
Spectral Resolution
Spectral Range
ImageResolution
Spatial Resolution
Temporal
Hyp
ersp
ectr
al-
imag
ing
Determinant Max. OPDΔδx + Camera QE
Camera speed + PZT speed
Diffraction-limitedCamera specs
Camera speedImage resolutionSpectral resolutionSNR requirement
Typical valueMax.~10cm-1
Tunable~360-900nm
Max.300x300pixels
500nm50x, NA=0.55
20s200x200pixels, 512frames
On
ly-i
mag
ing Determinant Camera QE Camera Pixel #
Diffraction-limitedCamera specs
Camera Speed
Typical value 200-1100nm1004x1002pixels (Cascade 1k, Photometrics)
500nm50x, NA=0.55
10fps@ full frame
Jianping. Li, Robert. Chan, and Xuzhu. Wang, Tests of a practical visible-NIR imaging Fourier transform spectrometer for biological and chemical fluorescence emission measurements. Opt. Express, 2009. 17(23).
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Instrumental Reconfigurability
Transmission Reflection
EPI-Fluorescence TIR-Fluorescence
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Prototype Gen2
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Absorption
0
( )( ) log[ ( )] log[ ]
( )I
A TI
Beer’s law:
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Reflection
Mixed SCCBs with red, green and blue color and common glass beads
JP. Li and Robert Chan. Towards a UV-Vis-NIR Hyperspectral Imaging Platform for Fast Multiplex Reflection Spectroscopy, Opt. Lett., 2010. 35(20): p. 3330-3332.
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502nm, 550nm and 630nm
JP. Li and Robert Chan. Towards a UV-Vis-NIR Hyperspectral Imaging Platform for Fast Multiplex Reflection Spectroscopy, Opt. Lett., 2010. 35(20): p. 3330-3332.
512 frames, 100×100pixels, binning =8. total acquisition time is only about 10s.
SCCB decoding speed of about 100beads/s.
Reflection: SCCBs
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Fluorescence
Li, J., R.K.Y. Chan, and X. Wang, Tests of a practical visible-NIR imaging Fourier transform spectrometer for biological and chemical fluorescence emission measurements. Opt. Express, 2009. 17(23).
50x, 100x100 pixels, binning=8, 5ms exposure time, 1k frames, total acquisition time ~40s
Dye-loaded fluorescent polystyrene nano-beads (φ200nm)
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TIRF
Jianping Li and Robert Chan, “Two-Mode Total Internal Reflection Fluorescence Hyperspectral Microscopy”, Focus on Microscopy, March, 2010, Shanghai.
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Towards a general-purpose Hyperspectral Microscopy System (Gen3)
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Design Considerations
Integration with standard microscopes1
Equip with faster and larger-chipped camera2
Equip with GPU based parallel FFT processing3
User-friendly software GUI development4
Versatile SI platform for analytical sciences5
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The real HSM system
Jianping Li and Yi Xiao, “GPU accelerated parallel FFT processing for Fourier transform hyperspectral imaging”, Applied Optics (IF: 1.784), 54(13): p. D91-D98, 2015.
SoftwareHardware
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Optical Physical
Performance and specs
• Spectral resolution (max):~0.4nm
• Spatial resolution (max):~0.5m
• Spectral range:~360-1000nm
• Pixel number: 2560x2160
• Instrument size: • 30×50×28mm (L×W×H)
• Objective lenses:• 5x,10x,20x,50x,80x BDF-RT-PL• 10x, 20x, 40x Pha
• Host computer: • Intel i7-3820CPU• NVIDIA GeForce GTX650 GPU• 64GB RAM• 2TB HDD
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ApplicationsBright-field (transmission)
Bright-field (phase-contrast)
Bright-field (reflection)
Dark-field (scattering)
Electroluminescence
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Technological Comparison
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FeaturesExcellent spatial & spectral resolving
High & adjustable spectral resolution and wide spectral range
High & adjustable spatial multiplex and resolution
No spatial resolution degradation
Good accuracy
High measurement throughput
High optical throughput
Rapid image acquisition
Parallel FFT processing
Operational
convenience
Multi-modality imaging
Flexible hardware adaptability
“WYSIWYG” preview function
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Acknowledgement• Dr. Robert Chan,
▫ Dr. XZ Wang
▫ Dr. JX Fu,
▫ Mr. R Li
▫ Mr. ZN Xu @HKBU
• Dr. XW Zhao and Prof. ZZ Gu @SEU
• Prof. JF Wang @CUHK
• Prof. B Ren @XMU