In order to evaluate the microvascular permeability of donor lungs, we calculated the filtration variables; K (reflection coefficient), simultaneously. For the calculation of filtration variables, the isolated rat lungs were kept in zone three conditions (pulmonary arterial pressure > pulmonary venous pressure > alveolar pressure). We used Krebs-Henseleit solution (K-H) with 6% bovine serum albumin for the perfusate, and maintained perfusion using a constant pressure circuit system. The pulmonary venous and alveolar pressures were maintained at 2.5 and 2.0 cmH2O, respectively, and pulmonary arterial pressure was set so that change in lung weight occurred. We increased both pulmonary arterial and venous pressures by 3 cmH2O simultaneously, and obtained the initial filtration rate. Then we diluted the perfusate with K-H to about half concentration, and obtained the initial filtration rate. We obtained values of filtration variables of K=4.55 mg.min-1.cm H2O-1.g-1], Ppmv=0.59 cmH2O, and sigma =0.617. These values agree with values from previous reports. Since these three filtration variables are interrelated, this method for simultaneous measurement is more accurate than independent measurements. The chief advantages of this method are that it does not require direct measurement of interstitial pressure or collection of lymph fluid, and all variables are obtained simultaneously. Using this method, the vascular permeability of isolated rat lungs, can be estimated.