Video-rate Raman-based metabolic imaging by Airy light-sheet illumination and photon-sparse detection

Dunn, Lochlann; Luo, Haokun; Subedi, Nava R.; Kasu, Ramachandran; McDonald, Armando G.; Christodoulides, Demetrios N.; Vasdekis, Andreas E.

doi:10.1073/pnas.2210037120

Video-rate Raman-based metabolic imaging by Airy light-sheet illumination and photon-sparse detection

Journal Article · Wed Feb 22 00:00:00 EST 2023 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.2210037120· OSTI ID:1958232

Dunn, Lochlann ^[1]; Luo, Haokun ^[2]; Subedi, Nava R. ^[1]; ^[1]; ^[3]; Christodoulides, Demetrios N. ^[2]; Vasdekis, Andreas E. ^[1]

Department of Physics, University of Idaho, Moscow, ID 83844-0903
The College of Optics and Photonics, University of Central Florida, Orlando, FL 32816-2700
Department of Forest, Rangeland, and Fire Sciences, University of Idaho, Moscow, ID 83844-1132

Despite its massive potential, Raman imaging represents just a modest fraction of all research and clinical microscopy to date. This is due to the ultralow Raman scattering cross-sections of most biomolecules that impose low-light or photon-sparse conditions. Bioimaging under such conditions is suboptimal, as it either results in ultralow frame rates or requires increased levels of irradiance. Here, we overcome this tradeoff by introducing Raman imaging that operates at both video rates and 1,000-fold lower irradiance than state-of-the-art methods. To accomplish this, we deployed a judicially designed Airy light-sheet microscope to efficiently image large specimen regions. Further, we implemented subphoton per pixel image acquisition and reconstruction to confront issues arising from photon sparsity at just millisecond integrations. We demonstrate the versatility of our approach by imaging a variety of samples, including the three-dimensional (3D) metabolic activity of single microbial cells and the underlying cell-to-cell variability. To image such small-scale targets, we again harnessed photon sparsity to increase magnification without a field-of-view penalty, thus, overcoming another key limitation in modern light-sheet microscopy.

View Journal Article

Research Organization:: Univ. of Idaho, Moscow, ID (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0019249; SC0022282

OSTI ID:: 1958232

Alternate ID(s):: OSTI ID: 1962095

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 9 Vol. 120; ISSN 0027-8424

Publisher:: Proceedings of the National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: English

References (60)

Phototoxicity in live fluorescence microscopy, and how to avoid it Icha, Jaroslav; Weber, Michael; Waters, Jennifer C. BioEssays, Vol. 39, Issue 8 https://doi.org/10.1002/bies.201700003	journal	July 2017
Microbial metabolic noise Vasdekis, Andreas E.; Singh, Abhyudai WIREs Mechanisms of Disease, Vol. 13, Issue 3 https://doi.org/10.1002/wsbm.1512	journal	November 2020
Progress in quantitative single-molecule localization microscopy Deschout, H.; Shivanandan, A.; Annibale, P. Histochemistry and Cell Biology, Vol. 142, Issue 1 https://doi.org/10.1007/s00418-014-1217-y	journal	April 2014
Nonresonant Confocal Raman Imaging of DNA and Protein Distribution in Apoptotic Cells Uzunbajakava, N.; Lenferink, A.; Kraan, Y. Biophysical Journal, Vol. 84, Issue 6 https://doi.org/10.1016/S0006-3495(03)75124-8	journal	June 2003
Raman-probes for monitoring metabolites and nutrient fate in Yarrowia lipolytica using deuterated glucose Kukal, Gurkeerat; Vasdekis, Andreas E.; McDonald, Armando G. Biocatalysis and Agricultural Biotechnology, Vol. 39 https://doi.org/10.1016/j.bcab.2021.102241	journal	January 2022
Cellular Metabolism and Disease: What Do Metabolic Outliers Teach Us? DeBerardinis, Ralph J.; Thompson, Craig B. Cell, Vol. 148, Issue 6 https://doi.org/10.1016/j.cell.2012.02.032	journal	March 2012
Fast interscale wavelet denoising of Poisson-corrupted images Luisier, Florian; Vonesch, Cédric; Blu, Thierry Signal Processing, Vol. 90, Issue 2 https://doi.org/10.1016/j.sigpro.2009.07.009	journal	February 2010
Cellular Imaging Using Stimulated Raman Scattering Microscopy Hill, Andrew H.; Fu, Dan Analytical Chemistry, Vol. 91, Issue 15 https://doi.org/10.1021/acs.analchem.9b02095	journal	June 2019
Probing Methionine Uptake in Live Cells by Deuterium Labeling and Stimulated Raman Scattering Spratt, Spencer J.; Oguchi, Kenichi; Miura, Keisuke The Journal of Physical Chemistry B, Vol. 126, Issue 8 https://doi.org/10.1021/acs.jpcb.1c08343	journal	February 2022
Photon-Sparse, Poisson Light-Sheet Microscopy Sanchez, Tristin; Subedi, Nava R.; Thompson, Cole ACS Photonics, Vol. 8, Issue 10 https://doi.org/10.1021/acsphotonics.1c01142	journal	September 2021
Two-Photon Fluorescence Microscopy at Extremely Low Excitation Intensity: The Power of Quantum Correlations Varnavski, Oleg; Goodson, Theodore Journal of the American Chemical Society, Vol. 142, Issue 30 https://doi.org/10.1021/jacs.0c01153	journal	July 2020
Studying single living cells and chromosomes by confocal Raman microspectroscopy Puppels, G. J.; de Mul, F. F. M.; Otto, C. Nature, Vol. 347, Issue 6290 https://doi.org/10.1038/347301a0	journal	September 1990
Scaling metabolism from organisms to ecosystems Enquist, Brian J.; Economo, Evan P.; Huxman, Travis E. Nature, Vol. 423, Issue 6940 https://doi.org/10.1038/nature01671	journal	June 2003
Multi-isotope imaging mass spectrometry quantifies stem cell division and metabolism Steinhauser, Matthew L.; Bailey, Andrew P.; Senyo, Samuel E. Nature, Vol. 481, Issue 7382 https://doi.org/10.1038/nature10734	journal	January 2012
The elusive Heisenberg limit in quantum-enhanced metrology Demkowicz-Dobrzański, Rafał; Kołodyński, Jan; Guţă, Mădălin Nature Communications, Vol. 3, Issue 1 https://doi.org/10.1038/ncomms2067	journal	January 2012
An entanglement-enhanced microscope Ono, Takafumi; Okamoto, Ryo; Takeuchi, Shigeki Nature Communications, Vol. 4, Issue 1 https://doi.org/10.1038/ncomms3426	journal	September 2013
Imaging with a small number of photons Morris, Peter A.; Aspden, Reuben S.; Bell, Jessica E. C. Nature Communications, Vol. 6, Issue 1 https://doi.org/10.1038/ncomms6913	journal	January 2015
NIH Image to ImageJ: 25 years of image analysis Schneider, Caroline A.; Rasband, Wayne S.; Eliceiri, Kevin W. Nature Methods, Vol. 9, Issue 7 https://doi.org/10.1038/nmeth.2089	journal	June 2012
Light-sheet microscopy using an Airy beam Vettenburg, Tom; Dalgarno, Heather I. C.; Nylk, Jonathan Nature Methods, Vol. 11, Issue 5 https://doi.org/10.1038/nmeth.2922	journal	April 2014
Eliciting the impacts of cellular noise on metabolic trade-offs by quantitative mass imaging Vasdekis, A. E.; Alanazi, H.; Silverman, A. M. Nature Communications, Vol. 10, Issue 1 https://doi.org/10.1038/s41467-019-08717-w	journal	February 2019
Super-resolution enhancement by quantum image scanning microscopy Tenne, Ron; Rossman, Uri; Rephael, Batel Nature Photonics, Vol. 13, Issue 2 https://doi.org/10.1038/s41566-018-0324-z	journal	December 2018
Quantum-enhanced nonlinear microscopy Casacio, Catxere A.; Madsen, Lars S.; Terrasson, Alex Nature, Vol. 594, Issue 7862 https://doi.org/10.1038/s41586-021-03528-w	journal	June 2021
Label-free metabolic imaging by mid-infrared optoacoustic microscopy in living cells Pleitez, Miguel A.; Khan, Asrar Ali; Soldà, Alice Nature Biotechnology, Vol. 38, Issue 3 https://doi.org/10.1038/s41587-019-0359-9	journal	December 2019
Highly-multiplexed volumetric mapping with Raman dye imaging and tissue clearing Shi, Lixue; Wei, Mian; Miao, Yupeng Nature Biotechnology, Vol. 40, Issue 3 https://doi.org/10.1038/s41587-021-01041-z	journal	October 2021
Biological imaging of chemical bonds by stimulated Raman scattering microscopy Hu, Fanghao; Shi, Lixue; Min, Wei Nature Methods, Vol. 16, Issue 9 https://doi.org/10.1038/s41592-019-0538-0	journal	August 2019
Mid-infrared metabolic imaging with vibrational probes Shi, Lixue; Liu, Xinwen; Shi, Lingyan Nature Methods, Vol. 17, Issue 8 https://doi.org/10.1038/s41592-020-0883-z	journal	June 2020
Advances in measuring cancer cell metabolism with subcellular resolution Ruiz-Rodado, Victor; Lita, Adrian; Larion, Mioara Nature Methods, Vol. 19, Issue 9 https://doi.org/10.1038/s41592-022-01572-6	journal	August 2022
Integrative quantitative-phase and airy light-sheet imaging Subedi, N. R.; Jung, P. S.; Bredeweg, E. L. Scientific Reports, Vol. 10, Issue 1 https://doi.org/10.1038/s41598-020-76730-x	journal	November 2020
The Potential of Metabolic Imaging Gialleonardo, Valentina Di; Wilson, David M.; Keshari, Kayvan R. Seminars in Nuclear Medicine, Vol. 46, Issue 1 https://doi.org/10.1053/j.semnuclmed.2015.09.004	journal	January 2016
Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy Evans, C. L.; Potma, E. O.; Puoris'haag, M. Proceedings of the National Academy of Sciences, Vol. 102, Issue 46 https://doi.org/10.1073/pnas.0508282102	journal	November 2005
Real-time metabolic imaging Golman, Klaes; in ‘t Zandt, René; Thaning, Mikkel Proceedings of the National Academy of Sciences, Vol. 103, Issue 30 https://doi.org/10.1073/pnas.0601319103	journal	July 2006
Monitoring of lipid storage in Caenorhabditis elegans using coherent anti-Stokes Raman scattering (CARS) microscopy Hellerer, T.; Axang, C.; Brackmann, C. Proceedings of the National Academy of Sciences, Vol. 104, Issue 37 https://doi.org/10.1073/pnas.0703594104	journal	September 2007
Label-free Raman observation of cytochrome c dynamics during apoptosis Okada, M.; Smith, N. I.; Palonpon, A. F. Proceedings of the National Academy of Sciences, Vol. 109, Issue 1 https://doi.org/10.1073/pnas.1107524108	journal	December 2011
Vibrational imaging of newly synthesized proteins in live cells by stimulated Raman scattering microscopy Wei, L.; Yu, Y.; Shen, Y. Proceedings of the National Academy of Sciences, Vol. 110, Issue 28 https://doi.org/10.1073/pnas.1303768110	journal	June 2013
Tracking heavy water (D ₂ O) incorporation for identifying and sorting active microbial cells Berry, David; Mader, Esther; Lee, Tae Kwon Proceedings of the National Academy of Sciences, Vol. 112, Issue 2 https://doi.org/10.1073/pnas.1420406112	journal	December 2014
Label-free DNA imaging in vivo with stimulated Raman scattering microscopy Lu, Fa-Ke; Basu, Srinjan; Igras, Vivien Proceedings of the National Academy of Sciences, Vol. 112, Issue 37 https://doi.org/10.1073/pnas.1515121112	journal	August 2015
Quantum imaging with sub-Poissonian light: challenges and perspectives in optical metrology Berchera, I. Ruo; Degiovanni, I. P. Metrologia, Vol. 56, Issue 2 https://doi.org/10.1088/1681-7575/aaf7b2	journal	January 2019
Quantum Optics: An Introduction Fox, Mark https://doi.org/10.1093/oso/9780198566724.001.0001	book	April 2006
Should we consider subcellular compartmentalization of metabolites, and if so, how do we measure them? Wellen, Kathryn E.; Snyder, Nathaniel W. Current Opinion in Clinical Nutrition & Metabolic Care, Vol. 22, Issue 5 https://doi.org/10.1097/MCO.0000000000000580	journal	September 2019
Observation of Accelerating Airy Beams Siviloglou, G. A.; Broky, J.; Dogariu, A. Physical Review Letters, Vol. 99, Issue 21 https://doi.org/10.1103/PhysRevLett.99.213901	journal	November 2007
Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy Huisken, J.; Swoger, J.; Del Bene, F. Science, Vol. 305, Issue 5686, p. 1007-1009 https://doi.org/10.1126/science.1100035	journal	August 2004
Challenges in Engineering Microbes for Biofuels Production Stephanopoulos, Gregory Science, Vol. 315, Issue 5813, p. 801-804 https://doi.org/10.1126/science.1139612	journal	February 2007
Manufacturing Molecules Through Metabolic Engineering Keasling, Jay D. Science, Vol. 330, Issue 6009, p. 1355-1358 https://doi.org/10.1126/science.1193990	journal	December 2010
Video-Rate Molecular Imaging in Vivo with Stimulated Raman Scattering Saar, B. G.; Freudiger, C. W.; Reichman, J. Science, Vol. 330, Issue 6009 https://doi.org/10.1126/science.1197236	journal	December 2010
First-Photon Imaging Kirmani, Ahmed; Venkatraman, Dheera; Shin, Dongeek Science, Vol. 343, Issue 6166 https://doi.org/10.1126/science.1246775	journal	November 2013
Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution Chen, Bi-Chang; Legant, Wesley R.; Wang, Kai Science, Vol. 346, Issue 6208 https://doi.org/10.1126/science.1257998	journal	October 2014
Coherent Anti-Stokes Raman Scattering Microscopy: Chemical Imaging for Biology and Medicine Evans, Conor L.; Xie, X. Sunney Annual Review of Analytical Chemistry, Vol. 1, Issue 1 https://doi.org/10.1146/annurev.anchem.1.031207.112754	journal	July 2008
Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy Minamikawa, Takeo; Murakami, Yoshinori; Matsumura, Naokazu Journal of Spectroscopy, Vol. 2017 https://doi.org/10.1155/2017/5725340	journal	January 2017
A molecular genetic toolbox for Yarrowia lipolytica Bredeweg, Erin L.; Pomraning, Kyle R.; Dai, Ziyu Biotechnology for Biofuels, Vol. 10, Issue 1 https://doi.org/10.1186/s13068-016-0687-7	journal	January 2017
Planar Airy beam light-sheet for two-photon microscopy Hosny, Neveen A.; Seyforth, James A.; Spickermann, Gunnar Biomedical Optics Express, Vol. 11, Issue 7 https://doi.org/10.1364/BOE.395547	journal	June 2020
Phototoxicity induced in living HeLa cells by focused femtosecond laser pulses: a data-driven approach Talone, B.; Bazzarelli, M.; Schirato, A. Biomedical Optics Express, Vol. 12, Issue 12 https://doi.org/10.1364/BOE.441225	journal	December 2021
Characterization of photodamage in coherent anti-Stokes Raman scattering microscopy Fu, Yan; Wang, Haifeng; Shi, Riyi Optics Express, Vol. 14, Issue 9 https://doi.org/10.1364/OE.14.003942	journal	January 2006
Light sheet-excited spontaneous Raman imaging of a living fish by optical sectioning in a wide field Raman microscope Oshima, Yusuke; Sato, Hidetoshi; Kajiura-Kobayashi, Hiroko Optics Express, Vol. 20, Issue 15 https://doi.org/10.1364/OE.20.016195	journal	January 2012
Photon-counting, energy-resolving and super-resolution phase contrast X-ray imaging using an integrating detector O’Connell, Dylan W.; Morgan, Kaye S.; Ruben, Gary Optics Express, Vol. 28, Issue 5 https://doi.org/10.1364/OE.384928	journal	January 2020
Airy light-sheet Raman imaging Subedi, N. R.; Yaraghi, S.; Jung, P. S. Optics Express, Vol. 29, Issue 20 https://doi.org/10.1364/OE.435293	journal	January 2021
Lock-in-detection-free line-scan stimulated Raman scattering microscopy for near video-rate Raman imaging Wang, Zi; Zheng, Wei; Huang, Zhiwei Optics Letters, Vol. 41, Issue 17 https://doi.org/10.1364/OL.41.003960	journal	January 2016
Spatial frequency modulated imaging in coherent anti-Stokes Raman microscopy Heuke, Sandro; Sivankutty, Siddharth; Scotte, Camille Optica, Vol. 7, Issue 5 https://doi.org/10.1364/OPTICA.386526	journal	May 2020
Advanced methods of microscope control using μManager software Edelstein, Arthur D.; Tsuchida, Mark A.; Amodaj, Nenad Journal of Biological Methods, Vol. 1, Issue 2 https://doi.org/10.14440/jbm.2014.36	journal	July 2014
Nonperturbative Chemical Imaging of Organelle Transport in Living Cells with Coherent Anti-Stokes Raman Scattering Microscopy Nan, Xiaolin; Potma, Eric O.; Xie, X. Sunney Biophysical Journal, Vol. 91, Issue 2 https://doi.org/10.1529/biophysj.105.074534	journal	July 2006
Transparent soil microcosms for live-cell imaging and non-destructive stable isotope probing of soil microorganisms Sharma, Kriti; Palatinszky, Márton; Nikolov, Georgi eLife, Vol. 9 https://doi.org/10.7554/eLife.56275	journal	November 2020

Similar Records

Related Subjects

60 APPLIED LIFE SCIENCES
light-sheet
metabolism
microscopy
single cell

Video-rate Raman-based metabolic imaging by Airy light-sheet illumination and photon-sparse detection

Citation Formats

References (60)

Similar Records

Related Subjects