Compact Image Slicing Spectrometer (ISS) for hyperspectral fluorescence microscopy.

An image slicing spectrometer (ISS) for microscopy applications is presented. Its principle is based on the redirecting of image zones by specially organized thin mirrors within a custom fabricated component termed an image slicer. The demonstrated prototype can simultaneously acquire a 140 nm spectral range within its 2D field of view from a single image. The spectral resolution of the system is 5.6 nm. The FOV and spatial resolution of the ISS depend on the selected microscope objective and for the results presented is 45 x 45 microm(2) and 0.45 microm respectively. This proof-of-concept system can be easily improved in the future for higher (both spectral and spatial) resolution imaging. The system requires no scanning and minimal post data processing. In addition, the reflective nature of the image slicer and use of prisms for spectral dispersion make the system light efficient. Both of the above features are highly valuable for real time fluorescent-spectral imaging in biological and diagnostic applications.

[1]  Linda T. Nieman,et al.  In vivo hyperspectral confocal fluorescence imaging to determine pigment localization and distribution in cyanobacterial cells , 2008, Proceedings of the National Academy of Sciences.

[2]  G. Bearman,et al.  Resolution of multiple green fluorescent protein color variants and dyes using two-photon microscopy and imaging spectroscopy. , 2001, Journal of biomedical optics.

[3]  H. E. Bennett,et al.  Infrared Reflectance of Evaporated Aluminum Films , 1962 .

[4]  W. Preuss,et al.  Precision machining of integral field units , 2006 .

[5]  Roland Bacon,et al.  Design of an Integral Field Unit for MUSE, and Results from Prototyping , 2006 .

[6]  Der Yi Hsu,et al.  Wide-range tunable Fabry-Perot array filter for wavelength-division multiplexing applications. , 2005, Applied optics.

[7]  Cornelis M. Dubbeldam,et al.  Prototyping of diamond machined optics for the KMOS and JWST NIRSpec integral field units , 2006, SPIE Astronomical Telescopes + Instrumentation.

[8]  David A. Landgrebe,et al.  Information Extraction Principles and Methods for Multispectral and Hyperspectral Image Data , 1999 .

[9]  Dario Cabib,et al.  Fourier transform multipixel spectroscopy for quantitative cytology , 1996 .

[10]  Hideaki Matsuoka,et al.  Single-cell viability assessment with a novel spectro-imaging system. , 2002, Journal of biotechnology.

[11]  Eric Prieto,et al.  Original image slicer designed for integral field spectroscopy with the near-infrared spectrograph for the James Webb Space Telescope , 2006 .

[12]  Roland Bacon,et al.  Slicing the universe at affordable cost: the quest for the MUSE image slicer , 2004, SPIE Optical Systems Design.

[13]  M E Gehm,et al.  Single-shot compressive spectral imaging with a dual-disperser architecture. , 2007, Optics express.

[14]  Ashwin A. Wagadarikar,et al.  Single disperser design for coded aperture snapshot spectral imaging. , 2008, Applied optics.

[15]  Jerilyn A. Timlin,et al.  Advanced imaging of multiple mRNAs in brain tissue using a custom hyperspectral imager and multivariate curve resolution , 2006, Journal of Neuroscience Methods.

[16]  J. Allington-Smith Basic principles of integral field spectroscopy , 2006 .

[17]  Xiaobai Sun,et al.  Video rate spectral imaging using a coded aperture snapshot spectral imager. , 2009, Optics express.

[18]  Vasilis Ntziachristos,et al.  Looking and listening to light: the evolution of whole-body photonic imaging , 2005, Nature Biotechnology.

[19]  R. Pepperkok,et al.  Spectral imaging and its applications in live cell microscopy , 2003, FEBS letters.

[20]  A S Belmont,et al.  Visualizing chromosome dynamics with GFP. , 2001, Trends in cell biology.

[21]  Scott A Mathews,et al.  Design and fabrication of a low-cost, multispectral imaging system. , 2008, Applied optics.

[22]  N. Thatte,et al.  3D: The next generation near-infrared imaging spectrometer , 1996 .

[23]  Thomas Ried,et al.  From Silencing to Gene Expression Real-Time Analysis in Single Cells , 2004, Cell.

[24]  Yasushi Hiraoka,et al.  Multispectral imaging fluorescence microscopy for living cells. , 2002, Cell structure and function.

[25]  M. Descour,et al.  Large-image-format computed tomography imaging spectrometer for fluorescence microscopy. , 2001, Optics express.

[26]  David M. Haaland,et al.  Multivariate curve resolution for hyperspectral image analysis: applications to microarray technology , 2003, SPIE BiOS.

[27]  M. Descour,et al.  Computed tomography-based spectral imaging for fluorescence microscopy. , 2001, Biophysical journal.