Jean B, Schmolz M W, Schöllhorn V G
Department of Ophthalmology, University of Tübingen, Germany.
Med Biol Eng Comput. 1992 Jul;30(4):CE17-20. doi: 10.1007/BF02446173.
Coupling a fluorochrome (e.g. fluorescein-isothiocyanate, FITC) to a molecule can enhance specific laser light absorption, thus leading to alteration or even destruction of the molecule itself. Therefore antibodies were labelled with FITC (absorption maximum 480 nm) and irradiated with laser light (488 nm) under various conditions. Inactivation of antibodies could only be achieved at the absorption maximum of FITC (as measured by direct and indirect immunofluorescence). Positive linear correlation exists between the amount of destruction and both exposure time and energy. Similar destructive effects were obtained when FITC-labelled peroxidase was irradiated. In these cases, enzyme activities measured by absorption photometry also showed a positive correlation to the total amount of energy transferred. Non-labelled proteins were not affected by irradiation. So we conclude that labelling of proteins with fluorochromes provides a highly specific means of selection of target molecules to be destroyed or inactivated. The method is based upon the laws of linear optics and is different from photodynamic or photochemical actions. The destructions observed are most likely caused by thermally induced changes of the molecules' tertiary structure.
将荧光染料（如异硫氰酸荧光素，FITC）与分子偶联可增强特定激光的吸收，从而导致分子本身发生改变甚至破坏。因此，在各种条件下，用FITC（最大吸收波长480nm）标记抗体并用激光（488nm）照射。只有在FITC的最大吸收波长处（通过直接和间接免疫荧光测量）才能实现抗体的失活。破坏量与照射时间和能量之间存在正线性相关。当照射FITC标记的过氧化物酶时也获得了类似的破坏效果。在这些情况下，通过吸收光度法测量的酶活性也与转移的总能量呈正相关。未标记的蛋白质不受照射影响。因此我们得出结论，用荧光染料标记蛋白质提供了一种高度特异性的选择要破坏或失活的靶分子的方法。该方法基于线性光学定律，不同于光动力或光化学作用。观察到的破坏很可能是由分子三级结构的热诱导变化引起的。
