Wigle Jeffrey C, Holwitt Eric A, Estlack Larry E, Noojin Gary D, Saunders Katharine E, Yakovlev Valdislav V, Rockwell Benjamin A
Human Effectiveness Directorate, Bioeffects Division, Optical Radiation Bioeffects Branch, 711th Human Performance Wing, 4141 Petroleum Road, JBSA Fort Sam Houston, Texas 78234-2644.
Human Effectiveness Directorate, Bioeffects Division, Molecular Bioeffects Branch, 711th Human Performance Wing, Wright-Patterson AFB, Ohio 45433-5707 and Department of Chemistry, United States Air Force Academy, Colorado Springs, Colorado 80840-6230.
J Biomed Opt. 2014 Jan;19(1):15008. doi: 10.1117/1.JBO.19.1.015008.
Data showing what appears to be nonthermal inactivation of M13 bacteriophage (M13), Tobacco mosaic virus, Escherichia coli (E. coli), and Jurkatt T-cells following exposure to 80-fs pulses of laser radiation have been published. Interest in the mechanism led to attempts to reproduce the results for M13 and E. coli. Bacteriophage plaque-forming and bacteria colony-forming assays showed no inactivation of the microorganisms; therefore, model systems were used to see what, if any, damage might be occurring to biologically important molecules. Purified plasmid DNA (pUC19) and bovine serum albumin were exposed to and analyzed by agarose gel electrophoresis (AGE) and polyacrylamide gel electrophoresis (PAGE), respectively, and no effect was found. DNA and coat proteins extracted from laser-exposed M13 and analyzed by AGE or PAGE found no effect. Raman scattering by M13 in phosphate buffered saline was measured to determine if there was any physical interaction between M13 and femtosecond laser pulses, and none was found. Positive controls for the endpoints measured produced the expected results with the relevant assays. Using the published methods, we were unable to reproduce the inactivation results or to show any interaction between ultrashort laser pulses and buffer/water, DNA, protein, M13 bacteriophage, or E. coli.
已发表的数据显示，在暴露于80飞秒激光辐射脉冲后，M13噬菌体（M13）、烟草花叶病毒、大肠杆菌（E. coli）和Jurkatt T细胞似乎出现了非热失活。对其机制的兴趣促使人们尝试重现M13和大肠杆菌的实验结果。噬菌体噬菌斑形成和细菌集落形成试验表明这些微生物没有失活；因此，使用模型系统来观察对生物重要分子可能产生的任何损伤。分别将纯化的质粒DNA（pUC19）和牛血清白蛋白暴露于琼脂糖凝胶电泳（AGE）和聚丙烯酰胺凝胶电泳（PAGE）并进行分析，未发现有影响。从激光照射后的M13中提取DNA和外壳蛋白，通过AGE或PAGE分析，未发现有影响。测量了M13在磷酸盐缓冲盐水中的拉曼散射，以确定M13与飞秒激光脉冲之间是否存在任何物理相互作用，未发现有相互作用。所测量终点的阳性对照在相关试验中产生了预期结果。使用已发表的方法，我们无法重现失活结果，也无法显示超短激光脉冲与缓冲液/水、DNA、蛋白质、M13噬菌体或大肠杆菌之间存在任何相互作用。
