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Supermultiplexed optical imaging and barcoding with engineered polyynes

Hu, Fanghao and Zeng, Chen and Long, Rong and Miao, Yupeng and Wei, Lu and Xu, Qizhi and Min, Wei (2018) Supermultiplexed optical imaging and barcoding with engineered polyynes. Nature Methods, 15 (3). pp. 194-200. ISSN 1548-7091. PMCID PMC5831481. https://resolver.caltech.edu/CaltechAUTHORS:20180608-125759681

[img] Image (JPEG) (Supplementary Figure 1 : UV-Vis absorption spectra of polyynes from 2-yne to 6-yne) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 2 : Frequency exploration of polyynes through conjugation elongation, end-capping variations and isotope doping) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 3 : Immuno-staining and SRS imaging of TOM20 (mitochondrial marker) and histone H2B (metaphase) in fixed HeLa cells with 4-yne conjugated antibodies) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 4 : 15-color imaging of live HeLa cells with super-multiplexed polyynes) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 5 : Live-cell SRS imaging of organelle-targeted polyynes and co-localization with fluorescent organelle markers in HeLa cells) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 6 : Live-cell SRS imaging of organelle-targeted polyynes in COS-7 cells) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 7 : Excellent photo-stability of polyynes in live cell imaging) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 8 : Minimal cytotoxicity of polyynes in live cells and phototoxicity of SRS lasers) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 9 : 10 representative spectral barcodes in polystyrene beads by spontaneous Raman microscope) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 10 : 2-D matrix cross-verification of spectral barcoding with polyynes) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 11 : Hyperspectral SRS imaging of encoded beads in live cells) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 12 : Fast organelle imaging in live cells with reduced laser power and shorter pixel dwell time) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 13 : Frequency encryption for identity security and anti-counterfeiting with polyynes) - Supplemental Material
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[img] PDF (Supplementary Figures 1–13, Supplementary Tables 1–3 and Supplementary Note 1) - Supplemental Material
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[img] PDF (Life Sciences Reporting Summary) - Supplemental Material
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Abstract

Optical multiplexing has a large impact in photonics, the life sciences and biomedicine. However, current technology is limited by a 'multiplexing ceiling' from existing optical materials. Here we engineered a class of polyyne-based materials for optical supermultiplexing. We achieved 20 distinct Raman frequencies, as 'Carbon rainbow', through rational engineering of conjugation length, bond-selective isotope doping and end-capping substitution of polyynes. With further probe functionalization, we demonstrated ten-color organelle imaging in individual living cells with high specificity, sensitivity and photostability. Moreover, we realized optical data storage and identification by combinatorial barcoding, yielding to our knowledge the largest number of distinct spectral barcodes to date. Therefore, these polyynes hold great promise in live-cell imaging and sorting as well as in high-throughput diagnostics and screening.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/nmeth.4578DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5831481/PubMed CentralArticle
https://rdcu.be/URofPublisherFree ReadCube access
ORCID:
AuthorORCID
Hu, Fanghao0000-0002-8659-4027
Wei, Lu0000-0001-9170-2283
Min, Wei0000-0003-2570-3557
Additional Information:© 2018 Macmillan Publishers Limited. Received: 23 August 2017. Accepted: 05 December 2017. Published: 15 January 2018. We are grateful for the discussion with L. Brus, Y. Shen and Z. Chen. W.M. acknowledges support from NIH Director's New Innovator Award (1DP2EB016573), R01 (EB020892), the US Army Research Office (W911NF-12-1-0594), and the Camille and Henry Dreyfus Foundation. Author Contributions: F.H. performed the spectroscopy, microscopy and biological studies and analyzed the data with the help of Y.M., L.W. and Q.X.; C.Z. performed the chemical synthesis together with R.L.; F.H. and W.M. conceived the concept; F.H., C.Z. and W.M. designed the experiments and wrote the manuscript with input from all authors. Competing interests: Columbia University has filed a patent application (US 62/540,953) based on this study. Code availability. The MATLAB code is available from the corresponding author upon request. Life Sciences Reporting Summary. Further information on experimental design is available in the Life Sciences Reporting Summary. Data availability. The data that support the findings of this study are provided in Supplementary Figures 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, Supplementary Tables 1–3 and Supplementary Note 1 and are available from the corresponding author upon request.
Funders:
Funding AgencyGrant Number
NIH1DP2EB016573
NIHR01 EB020892
Army Research Office (ARO)W911NF-12-1-0594
Camille and Henry Dreyfus FoundationUNSPECIFIED
Issue or Number:3
PubMed Central ID:PMC5831481
Record Number:CaltechAUTHORS:20180608-125759681
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180608-125759681
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:86924
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:08 Jun 2018 20:40
Last Modified:23 Oct 2019 20:56

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