Niggli Hugo J, Tudisco Salvatore, Privitera Giuseppe, Applegate Lee Ann, Scordino Agata, Musumeci Franco
BioFoton AG, Treyvaux, Switzerland.
J Biomed Opt. 2005 Mar-Apr;10(2):024006. doi: 10.1117/1.1899185.
Photobiological research in the last 30 yr has shown the existence of ultraweak photon emission in biological tissue, which can be detected with sophisticated photomultiplier systems. Although the emission of this ultraweak radiation, often termed biophotons, is extremely low in mammalian cells, it can be efficiently increased by ultraviolet light. Most recently it was shown that UV-A (330 to 380 nm) releases such very weak cell radiation in differentiated human skin fibroblasts. Based on these findings, a new and powerful tool in the form of UV-A-laser-induced biophotonic emission of cultured cells was developed with the intention to detect biophysical changes between carcinogenic and normal cells. With suspension densities ranging from 1 to 8 x 10(6) cells/mL, it was evident that an increase of the UV-A-laser-light induced photon emission intensity could be observed in normal as well as melanoma cells. Using this new detection procedure of ultraweak light emission, photons in cell suspensions as low as 100 microL could be determined, which is a factor of 100 lower compared to previous procedures. Moreover, the detection procedure has been further refined by turning off the photomultiplier system electronically during irradiation leading to the first measurements of induced light emission in the cells after less than 10 micros instead of 150 ms, as reported in previous procedures. This improvement leads to measurements of light bursts up 10(7) photons/s instead of several hundred as found with classical designs. Overall, we find decreasing induction ratings between normal and melanoma cells as well as cancer-prone and melanoma cells. Therefore, it turns out that this highly sensitive and noninvasive device enables us to detect high levels of ultraweak photon emission following UV-A-laser-induced light stimulation within the cells, which enables future development of new biophysical strategies in cell research.
过去30年的光生物学研究表明，生物组织中存在超微弱光子发射，这种发射可用精密的光电倍增管系统检测到。尽管这种通常被称为生物光子的超微弱辐射在哺乳动物细胞中的发射极其微弱，但它可被紫外线有效增强。最近有研究表明，UV-A（330至380纳米）能在分化的人皮肤成纤维细胞中释放这种非常微弱的细胞辐射。基于这些发现，开发了一种新的强大工具，即UV-A激光诱导培养细胞的生物光子发射，旨在检测致癌细胞与正常细胞之间的生物物理变化。当悬浮密度在1至8×10⁶个细胞/毫升范围内时，很明显在正常细胞和黑色素瘤细胞中都能观察到UV-A激光诱导的光子发射强度增加。使用这种新的超微弱光发射检测程序，可以测定低至100微升细胞悬液中的光子，这比以前的程序低100倍。此外，通过在照射期间以电子方式关闭光电倍增管系统，对检测程序进行了进一步优化，从而首次在不到10微秒而非先前程序所报告的150毫秒内测量细胞中的诱导光发射。这种改进使得能够测量高达10⁷个光子/秒的光脉冲，而不是传统设计中发现的几百个。总体而言，我们发现正常细胞与黑色素瘤细胞以及易患癌症细胞与黑色素瘤细胞之间的诱导率在降低。因此，事实证明，这种高度灵敏且非侵入性的设备使我们能够检测细胞内UV-A激光诱导光刺激后超微弱光子发射的高水平，这为细胞研究中新的生物物理策略的未来发展提供了可能。
