Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood.
Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood.
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DOI：
10.1364/boe.7.003449
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发表时间：
2016-09
影响因子：
3.4
通讯作者：
I. Saytashev;R. Glenn;Gabrielle A. Murashova;Sam Osseiran;D. Spence;C. Evans;M. Dantus
中科院分区：
文献类型：
--
作者：
I. Saytashev;R. Glenn;Gabrielle A. Murashova;Sam Osseiran;D. Spence;C. Evans;M. Dantus
在双光子激发荧光(TPEF)显微镜中，红细胞(RBC)由于其微弱的荧光信号而呈现为暗盘状。在这里，我们使用15fs800 nm脉冲用于TPEF，45fs1060 nm脉冲用于三光子激发荧光和三次谐波(THG)成像。在比较了其他预期的红细胞内源性荧光团：NADH、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的荧光。我们发现，TPEF和THG显微镜都可以用来非侵入性地检查红细胞的形态，而不会破坏储血袋。
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.