Autofluorescence spectroscopy of optically trapped cells.
Autofluorescence spectroscopy of optically trapped cells.
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光学捕获细胞的自发荧光光谱。
DOI：
10.1111/j.1751-1097.1995.tb09143.x
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发表时间：
1995
影响因子：
3.3
通讯作者：
Tromberg,BJ
中科院分区：
文献类型：
--
作者：
König,K;Liu,Y;Sonek,GJ;Berns,MW;Tromberg,BJ
在单光束梯度力光阱（“光镊”）中监测细胞自发荧光光谱，以探测近红外和UVA（320-400 nm）微照射的生理效应。在捕获前，中国仓鼠卵巢细胞显示出弱的UVA激发的自发荧光，在455 nm处具有最大值，这是β-烟酰胺腺嘌呤二核苷酸（磷酸盐）发射的特征。在捕获过程中，即使功率密度高达70 MW/cm 2和辐射暴露量为100 GJ/cm 2，1064 nm NIR微束对荧光强度和光谱特性也没有强烈的影响。与1064 nm陷阱相反，760 nm捕获光束在5分钟内引起两倍自发荧光增加（约20 GJ/cm 2）。在1064 nm捕获期间暴露于365 nm UVA（1 W/cm 2）显著改变了细胞自发荧光，在10 min内导致与初始水平相比增加5倍和6 nm红移。我们的结论是，1064 nm的微束可以应用在一个较长的时间内，而不会产生自发荧光的变化特征的细胞氧化还原状态的改变。然而，760 nm效应可能通过双光子吸收机制发生，其以类似于UVA暴露的方式改变氧化还原平衡并使细胞处于氧化应激状态。
Cellular autofluorescence spectra were monitored in a single‐beam gradient force optical trap (“optical tweezers”) in order to probe the physiological effects of near infrared and UVA (320–400 nm) microirradiation. Prior to trapping, Chinese hamster ovary cells exhibited weak UVA‐excited autofluorescence with maxima at 455 nm characteristic of β‐nicotinamide adenine dinucleotide (phosphate) emission. No strong effect of a 1064 nm NIR microbeam on fluorescence intensity and spectral characteristics was found during trapping, even for power densities up to 70 MW/cm2and radiant exposures of 100 GJ/cm2. In contrast to the 1064 nm trap, a 760 nm trapping beam caused a two‐fold autofluorescence increase within 5 min (about 20 GJ/cm2). Exposure to 365 nm UVA (1 W/cm2) during 1064 nm trapping significantly altered cellular autofluorescence, causing, within 10 min, a five‐fold increase and a 6 nm red shiftversusinitial levels. We conclude that 1064 nm microbeams can be applied for an extended period without producing autofluorescence changes characteristic of alterations in the cellular redox state. However, 760 nm effects may occurviaa two‐photon absorption mechanism, which, in a manner similar to UVA exposure, alters the redox balance and places the cell in a state of oxidative stress.