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Saytashev I, Glenn R, Murashova GA, et al. Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood. Biomed Opt Express. 2016;7(9):3449-3460doi: 10.1364/BOE.7.003449.
Saytashev, I., Glenn, R., Murashova, G. A., Osseiran, S., Spence, D., Evans, C. L., & Dantus, M. (2016). Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood. Biomedical optics express, 7(9), 3449-3460. https://doi.org/10.1364/BOE.7.003449
Saytashev, Ilyas, et al. "Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood." Biomedical optics express vol. 7,9 (2016): 3449-3460. doi: https://doi.org/10.1364/BOE.7.003449
Saytashev I, Glenn R, Murashova GA, Osseiran S, Spence D, Evans CL, Dantus M. Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood. Biomed Opt Express. 2016 Aug 12;7(9):3449-3460. doi: 10.1364/BOE.7.003449. eCollection 2016 Sep 01. PMID: 27699111; PMCID: PMC5030023.
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Biomed Opt Express. 2016 Aug 12;7(9):3449-3460. doi: 10.1364/BOE.7.003449. eCollection 2016 Sep 01.

Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood.

Biomedical optics express

Ilyas Saytashev, Rachel Glenn, Gabrielle A Murashova, Sam Osseiran, Dana Spence, Conor L Evans, Marcos Dantus

Affiliations

  1. Department of Chemistry, Michigan State University, 578 S Shaw Ln., East Lansing, MI 48824, USA.
  2. Harvard-MIT Division of Health Sciences and Technology, 77 Massachusetts Avenue E25-519, Cambridge, MA 02139, USA; Wellman Center for Photomedicine, Harvard Medical School, Massachusetts General Hospital, 149 13th Street, Charlestown, MA 02129, USA.
  3. Wellman Center for Photomedicine, Harvard Medical School, Massachusetts General Hospital, 149 13th Street, Charlestown, MA 02129, USA.
  4. Department of Chemistry, Michigan State University, 578 S Shaw Ln., East Lansing, MI 48824, USA; Department of Physics and Astronomy, Michigan State University, 567 Wilson Rd., East Lansing, MI 48824, USA.

PMID: 27699111 PMCID: PMC5030023 DOI: 10.1364/BOE.7.003449

Abstract

Red blood cells (RBC) in two-photon excited fluorescence (TPEF) microscopy usually appear as dark disks because of their low fluorescent signal. Here we use 15fs 800nm pulses for TPEF, 45fs 1060nm pulses for three-photon excited fluorescence, and third harmonic generation (THG) imaging. We find sufficient fluorescent signal that we attribute to hemoglobin fluorescence after comparing time and wavelength resolved spectra of other expected RBC endogenous fluorophores: NADH, FAD, biliverdin, and bilirubin. We find that both TPEF and THG microscopy can be used to examine erythrocyte morphology non-invasively without breaching a blood storage bag.

Keywords: (180.0180) Microscopy; (180.4315) Nonlinear microscopy; (180.5810) Scanning microscopy

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