GSNO alters human fibrin fiber density and fiber diameter.

Platelet-poor plasma was incubated with GSNO as described in methods. Images of native fibrin clots were acquired using multiphoton microscopy and analyzed using custom designed computer software. Panels A,B,C,G show fibrin clot “read out” images at 0,1,2.5,3.75 mM GSNO, respectively. GSNO decreased fibrin fiber density, panel D, but increased fiber diameter to a maximum at 1 mM GSNO, panel E. Panel F plots fibrin density against fiber diameter. Panel C contained abnormal fibrin clusters with numerous thin diameter fibers protruding from a high intensity core. They decreased the average fiber diameter and shifted the expected density-diameter relationship panel F, point C (2.5 mM GSNO). Fibrin agglomerates were detected at the highest GSNO concentration tested, panel G. Fibrin clot parameters displayed on clot “read out” images: n (number of fibers), fiber density (fibers/100 um), fiber intensity (au, mean and CV), fiber diameter (nm, mean or median and CV), fiber thin-thick ratio vs control (diam-TTR (ctrl)), void area (%), crossing fibers (xfibers/fiber), trifunctional junctions (xbranching/fiber), fibrinogen clusters (FbgClusters/mm2) and fibrin agglomerates (AG/mm2). *p<0.05 vs control. Area is 59×59 um2. Colour bars are image intensity. Scale bars are 8.5 um.