title: Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood.
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.
authors.affiliation: Department of Chemistry, Michigan State University, 578 S Shaw Ln., East Lansing, MI 48824, USA.
authors.forename: Ilyas
authors.initials: I
authors.lastname: Saytashev
authors.affiliation: Department of Chemistry, Michigan State University, 578 S Shaw Ln., East Lansing, MI 48824, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Glenn
authors.affiliation: Department of Chemistry, Michigan State University, 578 S Shaw Ln., East Lansing, MI 48824, USA.
authors.forename: Gabrielle A
authors.initials: GA
authors.lastname: Murashova
authors.affiliation: 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.
authors.forename: Sam
authors.initials: S
authors.lastname: Osseiran
authors.affiliation: Department of Chemistry, Michigan State University, 578 S Shaw Ln., East Lansing, MI 48824, USA.
authors.forename: Dana
authors.initials: D
authors.lastname: Spence
authors.affiliation: Wellman Center for Photomedicine, Harvard Medical School, Massachusetts General Hospital, 149 13th Street, Charlestown, MA 02129, USA.
authors.forename: Conor L
authors.initials: CL
authors.lastname: Evans
authors.affiliation: 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.
authors.forename: Marcos
authors.initials: M
authors.lastname: Dantus
keywords: (180.0180) Microscopy; (180.4315) Nonlinear microscopy; (180.5810) Scanning microscopy