Intravital fluorescence imaging of mouse brain using implantable semiconductor devices and epi-illumination of biological tissue.

The application of the fluorescence imaging method to living animals, together with the use of genetically engineered animals and synthesized photo-responsive compounds, is a powerful method for investigating brain functions. Here, we report a fluorescence imaging method for the brain surface and deep brain tissue that uses compact and mass-producible semiconductor imaging devices based on complementary metal-oxide semiconductor (CMOS) technology. An image sensor chip was designed to be inserted into brain tissue, and its size was 1500 × 450 μm. Sample illumination is also a key issue for intravital fluorescence imaging. Hence, for the uniform illumination of the imaging area, we propose a new method involving the epi-illumination of living biological tissues, and we performed investigations using optical simulations and experimental evaluation.

[1]  T. Yatagai,et al.  Direct Fabrication of Surface Relief Holographic Diffusers in Azobenzene Polymer Films , 2005 .

[2]  D T Delpy,et al.  The use of the Henyey–Greenstein phase function in Monte Carlo simulations in biomedical optics , 2006, Physics in medicine and biology.

[3]  J. Ohta,et al.  A CMOS image sensor with low fixed pattern noise suitable for lensless observation system of digital enzyme-linked immunosorbent assay (ELISA) , 2013, 2013 IEEE International Meeting for Future of Electron Devices, Kansai.

[4]  J. Tulip,et al.  Monte Carlo modelling of angular radiance in tissue phantoms and human prostate: PDT light dosimetry. , 1997, Physics in medicine and biology.

[5]  A. Gamal,et al.  Miniaturized integration of a fluorescence microscope , 2011, Nature Methods.

[6]  K. Deisseroth,et al.  Circuit-breakers: optical technologies for probing neural signals and systems , 2007, Nature Reviews Neuroscience.

[7]  W. Webb,et al.  Nonlinear magic: multiphoton microscopy in the biosciences , 2003, Nature Biotechnology.

[8]  Jun Ohta,et al.  Multimodal Complementary Metal–Oxide–Semiconductor Sensor Device for Imaging of Fluorescence and Electrical Potential in Deep Brain of Mouse , 2010 .

[9]  W. Whang,et al.  Optical diffusers based on silicone emulsions , 2009 .

[10]  Teruo Okano,et al.  Vascularization in 3D tissue using cell sheet technology. , 2013, Regenerative medicine.

[12]  G. Ellis‐Davies,et al.  In vivo two‐photon uncaging of glutamate revealing the structure–function relationships of dendritic spines in the neocortex of adult mice , 2011, The Journal of physiology.

[13]  GeunHyung Kim,et al.  A PMMA composite as an optical diffuser in a liquid crystal display backlighting unit (BLU) , 2005 .

[14]  Laurie D. Burns,et al.  High-speed, miniaturized fluorescence microscopy in freely moving mice , 2008, Nature Methods.

[15]  Sooyoung Chung,et al.  Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex , 2005, Nature.

[16]  Wafik El-Deiry,et al.  Noninvasive vascular imaging in fluorescent tumors using multispectral unmixing. , 2008, BioTechniques.

[17]  K. Eliceiri,et al.  Goniometric measurements of thick tissue using Monte Carlo simulations to obtain the single scattering anisotropy coefficient , 2012, Biomedical optics express.

[18]  Jun Ohta,et al.  Complementary Metal Oxide Semiconductor Based Multimodal Sensor for In vivo Brain Function Imaging with a Function for Simultaneous Cell Stimulation , 2010 .

[19]  Euiheon Chung,et al.  In vivo wide-area cellular imaging by side-view endomicroscopy , 2010, Nature Methods.

[20]  Seok Hyun Yun,et al.  Light-guiding hydrogels for cell-based sensing and optogenetic synthesis in vivo , 2013, Nature Photonics.

[21]  Yasushi Miyashita,et al.  Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons , 2001, Nature Neuroscience.

[22]  Hiroaki Takehara,et al.  Lab-on-a-brain: Implantable micro-optical fluidic devices for neural cell analysis in vivo , 2014, Scientific Reports.

[23]  Damian J. Wallace,et al.  Single-spike detection in vitro and in vivo with a genetic Ca2+ sensor , 2008, Nature Methods.

[24]  Masahiro Nunoshita,et al.  On-chip biofluorescence imaging inside a brain tissue phantom using a CMOS image sensor for in vivo brain imaging verification , 2006 .

[25]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[26]  L. C. Henyey,et al.  Diffuse radiation in the Galaxy , 1940 .

[27]  Yaniv Ziv,et al.  Time-lapse imaging of disease progression in deep brain areas using fluorescence microendoscopy , 2011, Nature Medicine.

[28]  N. Katunuma,et al.  Reverse zymography using fluorogenic substrates for protease inhibitor detection , 2005, Electrophoresis.

[29]  C. Hsu,et al.  Fabrication of microlens array diffuser films with controllable haze distribution by combination of breath figures and replica molding methods. , 2008, Optics express.

[30]  Feng Zhang,et al.  An optical neural interface: in vivo control of rodent motor cortex with integrated fiberoptic and optogenetic technology , 2007, Journal of neural engineering.

[31]  K. Svoboda,et al.  Sparse optical microstimulation in barrel cortex drives learned behaviour in freely moving mice , 2008, Nature.

[32]  Karl Deisseroth,et al.  Optogenetics in Neural Systems , 2011, Neuron.

[33]  E. Cocker,et al.  Fiber-optic fluorescence imaging , 2005, Nature Methods.

[34]  Jun Ohta,et al.  One-chip sensing device (biomedical photonic LSI) enabled to assess hippocampal steep and gradual up-regulated proteolytic activities , 2008, Journal of Neuroscience Methods.

[35]  Jun Ohta,et al.  An implantable CMOS device for blood-flow imaging during experiments on freely moving rats , 2014 .

[36]  W. Denk,et al.  Deep tissue two-photon microscopy , 2005, Nature Methods.

[37]  Igor L. Medintz,et al.  Quantum Dots in Bioanalysis: A Review of Applications across Various Platforms for Fluorescence Spectroscopy and Imaging , 2013, Applied spectroscopy.

[38]  Ravindran Girija Aswathy,et al.  Near-infrared quantum dots for deep tissue imaging , 2010, Analytical and bioanalytical chemistry.

[39]  H Moseley,et al.  Monte Carlo simulations for optimal light delivery in photodynamic therapy of non-melanoma skin cancer , 2012, Physics in medicine and biology.

[40]  A. Borst,et al.  A genetically encoded calcium indicator for chronic in vivo two-photon imaging , 2008, Nature Methods.

[41]  Jun Ohta,et al.  Implantable CMOS Biomedical Devices , 2009, Sensors.