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      Super-multiplexed optical imaging and barcoding with engineered polyynes

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          Abstract

          Optical multiplexing impacts widely in photonics, life science, biomedicine and engineering. Despite intensive efforts, current technology is limited by a longstanding “multiplexing ceiling” from existing optical materials. Here we engineered a novel class of polyyne-based materials for optical super-multiplexing. 20 distinct Raman frequencies are achieved as “ Carbon rain bow” through rational engineering of conjugation length, bond-selective isotope doping and end-capping substitution of polyynes. With further probe functionalization, we demonstrated unprecedented 10-color organelle imaging in single living cell with high specificity, sensitivity, and photo-stability. Moreover, optical data storage and identification are realized by combinatorial barcoding, yielding the largest number of distinct spectral barcodes to date. Therefore, these versatile polyynes hold great promises in live-cell imaging and sorting, high-throughput diagnostics and screening, and information technology.

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          Most cited references37

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          Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy.

          Label-free chemical contrast is highly desirable in biomedical imaging. Spontaneous Raman microscopy provides specific vibrational signatures of chemical bonds, but is often hindered by low sensitivity. Here we report a three-dimensional multiphoton vibrational imaging technique based on stimulated Raman scattering (SRS). The sensitivity of SRS imaging is significantly greater than that of spontaneous Raman microscopy, which is achieved by implementing high-frequency (megahertz) phase-sensitive detection. SRS microscopy has a major advantage over previous coherent Raman techniques in that it offers background-free and readily interpretable chemical contrast. We show a variety of biomedical applications, such as differentiating distributions of omega-3 fatty acids and saturated lipids in living cells, imaging of brain and skin tissues based on intrinsic lipid contrast, and monitoring drug delivery through the epidermis.
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            Nanoparticles with Raman spectroscopic fingerprints for DNA and RNA detection.

            Multiplexed detection of oligonucleotide targets has been performed with gold nanoparticle probes labeled with oligonucleotides and Raman-active dyes. The gold nanoparticles facilitate the formation of a silver coating that acts as a surface-enhanced Raman scattering promoter for the dye-labeled particles that have been captured by target molecules and an underlying chip in microarray format. The strategy provides the high-sensitivity and high-selectivity attributes of gray-scale scanometric detection but adds multiplexing and ratioing capabilities because a very large number of probes can be designed based on the concept of using a Raman tag as a narrow-band spectroscopic fingerprint. Six dissimilar DNA targets with six Raman-labeled nanoparticle probes were distinguished, as well as two RNA targets with single nucleotide polymorphisms. The current unoptimized detection limit of this method is 20 femtomolar.
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              Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules.

              Multicolor optical coding for biological assays has been achieved by embedding different-sized quantum dots (zinc sulfide-capped cadmium selenide nanocrystals) into polymeric microbeads at precisely controlled ratios. Their novel optical properties (e.g., size-tunable emission and simultaneous excitation) render these highly luminescent quantum dots (QDs) ideal fluorophores for wavelength-and-intensity multiplexing. The use of 10 intensity levels and 6 colors could theoretically code one million nucleic acid or protein sequences. Imaging and spectroscopic measurements indicate that the QD-tagged beads are highly uniform and reproducible, yielding bead identification accuracies as high as 99.99% under favorable conditions. DNA hybridization studies demonstrate that the coding and target signals can be simultaneously read at the single-bead level. This spectral coding technology is expected to open new opportunities in gene expression studies, high-throughput screening, and medical diagnostics.
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                Author and article information

                Journal
                101215604
                32338
                Nat Methods
                Nat. Methods
                Nature methods
                1548-7091
                1548-7105
                21 December 2017
                15 January 2018
                March 2018
                15 July 2018
                : 15
                : 3
                : 194-200
                Affiliations
                [1 ]Department of Chemistry, Columbia University, New York, NY 10027, USA
                [2 ]Kavli Institute for Brain Science, Columbia University, New York, NY 10027, USA
                Author notes
                [* ]Corresponding author. E-mail: wm2256@ 123456columbia.edu
                [†]

                These authors contribute equally to this work

                Article
                NIHMS927119
                10.1038/nmeth.4578
                5831481
                29334378
                5cf4c948-4809-429f-8486-3c9ddb63cfae

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                Life sciences
                Life sciences

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