The objective is to quantify the guidewire (diameter of ) flow-obstruction effect in the in vitro model coronary stenoses in relation to trans-stenotic pressure drop, , fractional flow reserve (gFFR; “g” represents FFR measurement with guidewire insertion) and coronary flow reserve (gCFR) for steady and pulsatile physiological flows. The sensor tipped pressure or flow measuring guidewire insertion through stenotic lumen increases the trans-stenotic pressure drop or reduces the pharmacologically induced hyperemic flow in the coronary arteries with plaques. These hemodynamic changes may cause error in true FFR and CFR measurements, especially for intermediate coronary stenosis. To quantify guidewire flow-obstruction effect, simultaneous measurements of trans-stenotic pressures and flow were performed by two methods: (a) guidewire based measurements (gCFR and gFFR by inserting sensor tipped guidewire) and (b) true physiological measurements (CFR by in-line Doppler flow cuff and FFR by the radially drilled pressure ports in three epicardial coronary stenotic test sections, postangioplasty, intermediate, and preangioplasty). The diagnostic parameters measured before guidewire insertion (CFR and FFR) and during guidewire insertion (gCFR and gFFR) were validated numerically and correlated with the new diagnostic parameter “lesion flow coefficient (LFC).” There was significant flow reduction with increased trans-stenotic pressure drop due to guidewire insertion. The FFR-gFFR and CFR-gCFR correlations were and , respectively, where gCFR is reported from clinical pressure-flow data. Similar highly regressed correlations were obtained for LFC and gLFC with flow ratios and pressure ratios. There was significant difference between steady and pulsatile pressure drops for the same mean flow with and without guidewire insertion. The trans-stenotic hemodynamics was altered due to guidewire insertion. The true FFR and CFR were underestimated because of guidewire insertion. Hence, the FFR-gFFR and CFR-gCFR correlations can be used to find out true FFR and CFR from clinically measured values (i.e., gFFR and gCFR). In addition, the gLFC-gCFR and gLFC-gFFR were correlated significantly for post- and preangioplasty conditions.