Reflectance changes in clotting native blood: evidence of a red-cell process

When broadband light illuminates clotting native blood, the reflectance at each wavelength traces a time course with four discernible regions. Clot formation occurs just before the second phase. Two wavelengths, 471 and 771 nm, were selected for more detailed study of the first two phases. Analysis of each time course in native blood demonstrates that both signals track a single process during the first phase, but distinct processes during the second. Experiments on citrated blood identified which blood components contribute to reflectance changes. Comparison of liquid and clotting blood reveals a single process during the first phase, entailing that rouleaux formation determines the time course at both wavelengths. Control experiments eliminate clot propagation and shape change of red cells or platelets as possible factors in the second phase. Exogenous ADP added to EDTA blood evokes the second-phase response at 471 but not 771 nm, a novel phenomenon that requires the presence of red cells. The descriptive name 'ADP-end-response' is suggested for this red cell process until it is further characterized. We propose that the ADP-end-response determines the 471-nm signal during the second phase of clotting native blood and depends upon platelets in the absence of exogenous ADP. The 771-nm signal reports fibrin cross-linking during the second phase. An earlier pilot study demonstrated that rofecoxib effects the 471-nm signal ex vivo, which indicates that reflectance spectroscopy may be useful in the assessment of drug effects on platelet-erythrocyte interactions.