To uncover the pathological mechanism of neurodegenerative diseases, simple and cost-effective analytical methods are highly desirable to obtain biochemistry and morphology information on Tau protein aggregation. Here, we demonstrate the fluorescence-guided computational mid-infrared photothermal microscope, termed Fluorescence-guided Bond-selective Intensity Diffraction Tomography (FBS-IDT). FBS-IDT enables hyperspectral 3D bond-selective refractive index map recovery on Tau fibrils in cells. Depth-resolved mid-infrared fingerprint spectra extraction and related secondary protein structure analysis are demonstrated. 3D reconstruction of β sheet in Tau fibril is realized mainly based on the mid-infrared photothermal effects.
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