Time-resolved spectroscopy of hemoglobin and myoglobin in resting and ischemic muscle.

Difficulties of quantitation of hemoglobin/myoglobin absorption changes in muscle have led to the development of a new approach using short pulses of light. This method uses input light pulses sufficiently short so that the time course of travel of light through the brain can be precisely measured. The time of arrival of light at the detector gives the optical path length, given the velocity of light in tissues. The intensity profile of photon migration in tissues permits determination of the path length that the exiting photons have traveled and the concentration change of the pigments. A cavity-dumped liquid dye laser illuminates the tissue with 130-ps pulses detected as 600-ps duration at a half height at 3.0-cm distance from the input point. The decay of intensity from the 50% point onward to 0.1% follows a logarithmic function of slope mu which is attributed to the total absorption coefficient of the tissue. Increments of mu due to deoxyhemoglobin absorption at 760 and 630 nm are used to calculate the concentration change. This permits the calculation of the path length for continuous light measurements of 2 cm for a particular geometry. Variation of the wavelength of the laser affords determination of a spectrum of changes in the tissue.