Download Methods in enzimology v278 - Fluorescence Spectroscopy MIE by Brand, Johnson PDF
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Extra info for Methods in enzimology v278 - Fluorescence Spectroscopy MIE
Sample text
Problem: A dead-time problem occurs when t, is less than the read time associated with getting all of the data out of region 2 of the MCA. The least amount of time in which we can get 4000 channels of data out of the (up to) 8 MCAs (and still maintain all of the other duties of the instrument controller) is approximately 5-10 msec. This means that as we collect data on the sub-10-msec time scale, the time . 34 FLUORESCENCE SPECTROSCOPY [3] axis that is sampled contains dead time between the points (a solution to this problem is given in point 6).
It is important to recognize the two time scales associated with these experiments. The refolding reaction is being initiated utilizing standard stopped-flow technology with millisecond timing resolution. The light source utilized to obtain the fluorescence signal, however, instead of being continuous, is pulsed, at a repetition rate of approximately 4 million times per second (4 MHz, - 1 psec/pulse). Therefore, within any single 1-msec period, the laser will have excited the sample 4000 times.
Yoshihara, Z Phys. Chem. 95, 6095. 64 T. Kajiwara, R. W. Chambers, and D. R. Kearns, Chem. Phys. Lett. 22, 37 (1973). 22 FLUORESCENCE SPECTROSCOPY [2] absorption spectrum and in a strong decrease in the fluorescence quantum yield (fluorescence has its origin in the lowest excited level, the radiative transition from which to the ground state is forbidden in H-dimers). In contrast, in a J-dimer 012 = 01R = 02R = 0, which yields n = -2, and here the radiative transitions between the ground state and the upper excited state are forbidden, whereas transitions between the ground state and the lower excited state are enhanced twofold.