We demonstrated the potential of applying infrared spectroscopy for neuroscience by increasing both the spatial resolution and the specificity of chemical structure analysis in tissues and cells, thus overcoming the current limitations of conventional fluorescence microscopy and infrared microspectroscopy. Specifically, we used a novel instrumental setup that combines optical photothermal infrared spectroscopy (OPTIR) with an epifluorescent imaging module (EF) to allow IR spectroscopic analysis of fluorescently microglia cells surrounding amyloid plaques in comparison to microglial cells in amyloid-free tissue. We have demonstrated the capability of EF-OPTIR microscopy to locate specific cells and amyloid proteins and assess amyloid structures with sub-micron spatial resolution.
We have developed a new multimodal microscope enabling “Fluorescence-Guided” Optical Photothermal Infrared (O-PTIR) spectroscopy. O-PTIR has proven itself a breakthrough in IR spectroscopy, providing IR spectroscopic analysis with spatial resolution ~20x better than traditional IR in a non-contact optical configuration. We have now coupled O-PTIR with fluorescence microscopy, a long established, cornerstone technique in life science research. The new fluorescence guided O-PTIR instrument allows: (1) a sample to be analyzed via fluorescence microscopy to map sample regions that are labeled one or more fluorophores; and (2) use fluorescence images to direct infrared spectroscopic analysis in the vicinity of the fluorescently labeled regions. We have used fluorescence guided O-PTIR to perform spectroscopic analysis of the protein secondary structure of fluorescently labeled amyloid plaques in mouse brain tissue, with the goal of providing insights in neurodegenerative disease research.
The recent advent of Optical Photothermal IR (O-PTIR), has enabled for the first time, submicron infrared microscopy in far-field reflection mode with the combination of Raman for simultaneous, correlative IR+Raman microscopy. These unique and exciting synergistic capabilities are now spawning interest in life science application [1-2]. A broad range of life science applications, otherwise impossible with traditional FTIR/QCL microscopy, will be presented, ranging from live cell imaging in water, to ultra-high resolution images of breast tissue calcifications, amyloid aggregates in neurons (neurites and dendritic spines), individual collagen fibrils with polarized IR and individual isotopically labelled bacterial cells and more.
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