Steen H B, Sørensen O I
Department of Biophysics, Institute for Cancer Research, Oslo, Norway.
Cytometry. 1993;14(2):115-22. doi: 10.1002/cyto.990140203.
It has been found that the type of short arc high-pressure lamps used in some flow cytometers can be pulsed to reach intensities many times higher than that measured when they are operated on constant power. A 75 W mercury-xenon lamp was fed 20 microseconds current pulses super-imposed on its rated current of 5.4 A. Pulses of 50 A produced a 75-fold increase of the emission intensity at the excitation wavelength of FITC, which means that the excitation intensity in the flow chamber at this wavelength exceeded 100 mW. At the major emission lines of mercury, i.e., 366, 436, and 546 nm, the increase was about 25-fold, corresponding to intensities of the order of 300 mW. The light pulses were found to be reproducible to within 2% and the intensity was independent of the pulse frequency up to at least 250 pulses/s. Operated in this mode the lamp produced more than 1 x 10(8) 30 A pulses, which is sufficient to measure some 10,000 typical size samples. The rise time of the light pulses was about 5 microseconds. This was sufficiently short that the leading edge of the light scattering signal from a cell entering the excitation focus of the flow cytometer could be used to trigger the current pulse so that the cells were exposed to the full light pulse intensity while they were still within the focus.(ABSTRACT TRUNCATED AT 250 WORDS)
已发现一些流式细胞仪中使用的短弧高压灯类型可以被脉冲驱动,以达到比在恒定功率下运行时所测强度高出许多倍的强度。一个75瓦的汞 - 氙灯被施加叠加在其5.4安额定电流上的20微秒电流脉冲。50安的脉冲在FITC激发波长处使发射强度增加了75倍,这意味着在该波长下流动室中的激发强度超过了100毫瓦。在汞的主要发射线,即366、436和546纳米处,增加约为25倍,对应强度约为300毫瓦。发现光脉冲的再现性在2%以内,并且强度在至少250脉冲/秒的脉冲频率范围内与脉冲频率无关。以这种模式运行时,该灯产生了超过1×10⁸个30安的脉冲,这足以测量大约10000个典型大小的样本。光脉冲的上升时间约为5微秒。这足够短,以至于来自进入流式细胞仪激发焦点的细胞的光散射信号的前沿可用于触发电流脉冲,从而使细胞在仍处于焦点内时就暴露于全光脉冲强度下。(摘要截短于250字)