Pulse modulation of the excitation light source boosts the sensitivity of an arc lamp-based flow cytometer.
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)
Steen
H B
HB
Department of Biophysics, Institute for Cancer Research, Oslo, Norway.
Sørensen
O I
OI
D005434:Flow Cytometry / Q000295:instrumentation*; D008027:Light; D012680:Sensitivity and Specificity