Saytashev Ilyas, Glenn Rachel, Murashova Gabrielle A, Osseiran Sam, Spence Dana, Evans Conor L, Dantus Marcos
Department of Chemistry, Michigan State University, 578 S Shaw Ln., East Lansing, MI 48824, USA.
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.
Biomed Opt Express. 2016 Aug 12;7(9):3449-3460. doi: 10.1364/BOE.7.003449. eCollection 2016 Sep 1.
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.
在双光子激发荧光（TPEF）显微镜下，红细胞（RBC）由于其低荧光信号通常呈现为暗盘状。在这里，我们使用15飞秒800纳米脉冲进行TPEF，45飞秒1060纳米脉冲进行三光子激发荧光以及进行三次谐波产生（THG）成像。在比较了其他预期的红细胞内源性荧光团（烟酰胺腺嘌呤二核苷酸（NADH）、黄素腺嘌呤二核苷酸（FAD）、胆绿素和胆红素）的时间和波长分辨光谱后，我们发现了足够的荧光信号，我们将其归因于血红蛋白荧光。我们发现，TPEF和THG显微镜均可用于非侵入性检查红细胞形态，而无需刺破血袋。
