Raman microscopic imaging has emerged as an important research tool in life sciences because it does not require sample preparation and is non-destructive,non-invasive,and insensitive to aqueous solutions.It enables the characterization of biochemical components and sample distributions at a micrometer or nanometer scale.As the research on complex biological samples increases,Raman microscopic imaging holds potential for the dynamic stereoscopic observation of molecular composition and distribution in biological samples.This paper systematically examined recent advancements in 3D Raman microscopic imaging,including technical approaches,improvement strategies,and experimental results of various 3D imaging methods based on spontaneous Raman scattering,coherent Raman scattering,surface-enhanced Raman scattering,and Raman tags.Further,the progress of the applications of different imaging techniques in cell biology and developmental biology was summarized.Finally,the challenges and development prospects of different 3D Raman microscopic imaging techniques in the application of biomedical optical microscopic imaging technology were outlined.
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