We would like to measure the fluorescence decay of different gram positive bacteria using frequency domain instrumentation. In addition, we will investigate any differences in lifetime between vegetative cells and bacterial spores. The bacteria will not be tagged with a fluorescence dye so we will be looking at intrinsic fluorescence. Steady-state fluorescence of our target bacteria show excitation maxima at or around 290nm, 350nm, and 450nm. Therefore, we would like to excite our samples (suspended in di H2O) at these fluorescence excitation bands. The required instrumentation to resolve fluorescence lifetime of these multi-component samples is as follows: excitation wavelengths of 290nm 325nm, and 457nm; scan fluorescence emission from 10nm above the excitation wavelength to 20nm below the second order; and vary the modulation frequency from 10-200MHz using a 5MHz increment.
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