Carcinogenic damage to deoxyribonucleic acid is induced by near-infrared laser pulses in multiphoton microscopy via combination of two- and three-photon absorption
Carcinogenic damage to deoxyribonucleic acid is induced by near-infrared laser pulses in multiphoton microscopy via combination of two- and three-photon absorption
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多光子显微镜中的近红外激光脉冲通过两光子和三光子吸收的组合诱导脱氧核糖核酸的致癌损伤
DOI：
10.1117/1.jbo.17.11.116024
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发表时间：
2012
影响因子：
3.5
通讯作者：
H. Gerritsen
中科院分区：
文献类型：
--
作者：
O. Nadiarnykh;G. Thomas;Johan van Voskuilen;H. Sterenborg;H. Gerritsen
抽象的。应用于体内的非线性光学成像模式（多光子激发荧光，二次和三次谐波产生）越来越有希望用于临床诊断和监测癌症和其他疾病，因为它们可以在近红外（IR）波长下以高衍射极限分辨率探测组织。然而，双光子过程所需的飞秒激光脉冲的高峰强度导致细胞脱氧核糖核酸（DNA）中形成类似于暴露于太阳紫外（UV）光的损伤的环丁烷-嘧啶-二聚体（CPD）。后续突变的不准确修复增加了致癌的风险。在这项研究中，我们调查CPD的损害，结果在中国仓鼠卵巢细胞在体外成像与双光子激发自体荧光。CPD水平通过免疫荧光染色定量。我们进一步评估CPD损坏的程度，相对于不同的波长，在焦平面的脉冲宽度，和像素的停留时间相比，更明显的损害，从紫外线源。虽然预计CPD损伤是由三光子吸收引起的，但我们的结果表明，CPD是由竞争的双光子和三光子吸收过程引起的，其中前者进入UVA吸收带。这一发现是独立证实的激光功率，波长和脉冲宽度的损伤的非线性依赖关系。
Abstract. Nonlinear optical imaging modalities (multiphoton excited fluorescence, second and third harmonic generation) applied in vivo are increasingly promising for clinical diagnostics and the monitoring of cancer and other disorders, as they can probe tissue with high diffraction-limited resolution at near-infrared (IR) wavelengths. However, high peak intensity of femtosecond laser pulses required for two-photon processes causes formation of cyclobutane-pyrimidine-dimers (CPDs) in cellular deoxyribonucleic acid (DNA) similar to damage from exposure to solar ultraviolet (UV) light. Inaccurate repair of subsequent mutations increases the risk of carcinogenesis. In this study, we investigate CPD damage that results in Chinese hamster ovary cells in vitro from imaging them with two-photon excited autofluorescence. The CPD levels are quantified by immunofluorescent staining. We further evaluate the extent of CPD damage with respect to varied wavelength, pulse width at focal plane, and pixel dwell time as compared with more pronounced damage from UV sources. While CPD damage has been expected to result from three-photon absorption, our results reveal that CPDs are induced by competing two- and three-photon absorption processes, where the former accesses UVA absorption band. This finding is independently confirmed by nonlinear dependencies of damage on laser power, wavelength, and pulse width.
DOI：
10.1073/pnas.0502311102
发表时间：
2005-07-19
影响因子：
11.1
作者：
Besaratinia, A;Synold, TW;Pfeifer, GP
通讯作者：
Pfeifer, GP