Among various optical methods, fluorescence imaging has been the most widely exploited thanks to its superior sensitivity and specificity, but the resolvable colors are restricted to 2-5 colors because of the intrinsically broad and featureless spectra. Recently, this fluorescent “color barrier” was broken and super-multiplex optical imaging became possible taking advantage of well-designed Raman probes. However, the acquisition of the super-multiplex images is still relatively slow which impedes wider applications. Here, we demonstrate fast super-multiplex organelle imaging with high-speed color switching and acquisition, which accelerates the imaging speed by 2 orders of magnitude. We applied it in imaging cytometry, tracing mitosis and fast organelle motions in live cells. We anticipate that high-speed supermultiplex optical imaging can expand to a much wider field of biological researches.
Glucose is consumed as an energy source by virtually all living organisms, from bacteria to humans. Its uptake activity closely reflects the cellular metabolic status in various pathophysiological transformations, such as diabetes and cancer. Extensive efforts such as positron emission tomography, magnetic resonance imaging and fluorescence microscopy have been made to specifically image glucose uptake activity but all with technical limitations. Here, we report a new platform to visualize glucose uptake activity in live cells and tissues with subcellular resolution and minimal perturbation. A novel glucose analogue with a small alkyne tag (carbon-carbon triple bond) is developed to mimic natural glucose for cellular uptake, which can be imaged with high sensitivity and specificity by targeting the strong and characteristic alkyne vibration on stimulated Raman scattering (SRS) microscope to generate a quantitative three dimensional concentration map. Cancer cells with differing metabolic characteristics can be distinguished. Heterogeneous uptake patterns are observed in tumor xenograft tissues, neuronal culture and mouse brain tissues with clear cell-cell variations. Therefore, by offering the distinct advantage of optical resolution but without the undesirable influence of bulky fluorophores, our method of coupling SRS with alkyne labeled glucose will be an attractive tool to study energy demands of living systems at the single cell level.
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