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incubation is carried out in a moist 37°C incubator of time. A control slide (0 min=••incubation) is prepared for and to aid in interpretation of themsults. Incubations of 1) rinse slides in tap water bath 2) place in acid hematoxylin 90 sec. (or until red-blue color results 5) rinse in water • 6) 50% ethyl alcohol x 15 min. 7) 70•/G ethyl alcohol x 15 min. 8) 95% ethyl alcohol x 30 min. 9)10(/, ethyl alcohol x 45 min. 10) eosin Y x 20 sec. 11) wash in 100°; ethyl alcohol three times 12) 50% ethyl alcohol - 50% xylol 13) 100% xylol x 20 min. x 20 min. 14) Trim excess fibrim film from the slides with sharp knife and cover with synthetic mounting medium and a cover slip. The entire area under the cover slip must be filled with mountir.g meditnn to prevent drlrinh of the film. 'T};e sVes are •icw CoCi'JIIP.Int; S T-a 6 fe ) QuJ &_ILC~y 6,Z SyoreY; ~r_ot- The slides are best observecd witlh, a lm: pnwer stercoscopic micruscope. Lysed zones will first appear as depressedi ar.eps under the tissue section. 3) rinse in water 4) develop in dilute ammonia water until blue -*.Sei.vely inactive tissues have been carried :~r~ods of up to 24 hours. The reaction - 4 LCl% €ormaline-saline solution. : .~_ . ~: ..• . • .~. . ~ .' .' Staining procedure: 9

. . . . . . .:'~:'~J; may be made with reference to the time of incubation necessary : ~Semi-qeiontitative comparisons of activator activity in different ; ~ Sli~.fi iubated fc~r tonger pe : ~ds will have holes through the entire ri . . ..._ . . . .. . .. .. . . . "• -- • . .x, ..} fibrin film at areas under the tissue that correspond to the earlier, . . • #• - .. . _ . • ... . <; n ftresaions.-.Lysi:i an hea'ted_•films or those containing activator ,~ :~inhibitors ( e.q. K_x 10'~'i~AEA) is indicative of proteolytic activi in the tissc.e not due to plasminogen activator. ,.to procl~sce a certain observed amount of lysis. However, these comparisons mwat be made only between slides which are prepared with the same batch ". of fibrinogen on one film and with simultaneous incubations. It is preferable to use one batch of fibrinogen in which the plaminogen contamination is known (Ear assay in the true sense of the definition since the plasmin spreads. The Gh3~e~,=`-~xk~-: The method is not a histochemical localization is therefore dependent on a proper incubation time. Excessive-incubation of the slides results in autolysis of the entire tissue seetion:'';~:` 1