To explore the correlations between histopathologic findings and intravoxel incoherent motion (IVIM)-derived perfusion and diffusion parameters in brain gliomas. Thirty-two biopsy samples from twenty-one patients with newly diagnosed gliomas from a previous prospective cohort study were retrospectively analyzed. All patients underwent diffusion-weighted MRI with 22 b values (0-5000s/mm2), followed by intraoperative MR-guided biopsy surgery and surgical resection. All 32 biopsy samples underwent immunohistochemical staining followed by quantitative analysis of cell density (cellularity), percent of MIB-1 (Ki67)-positive expression (pMIB-1), number of CD34-stained vessels (CD34-MVD), and percent of VEGF-positive expressing cells (pVEGF) using a multispectral phenotyping microscope. Based on the co-registered localized biopsy, correlation analysis was performed between the IVIM-derived biexponential model-based parameters (Dfast1500 and Dfast5000, Dslow1500 and Dslow5000, PF1500 and PF5000) and the above four pathological biomarkers and glioma grades. Significant positive correlations were revealed between Dfast5000 and pVEGF (rho (r) = 0.466, p = 0.007), and Dfast1500 and pVEGF (r = 0.371, p = 0.037). A significant negative correlation was revealed between PF5000 with pMIB-1 (r = - 0.456, p = 0.01). Moderate to good positive correlations were shown between Dfast5000 and glioma grades (r = 0.509, p = 0.003) and Dfast1500 and glioma grades (r = 0.476, p = 0.006). IVIM-DWI-derived Dfast and PF correlate, respectively, with intratumor pVEGF and pMIB-1. When using the wide-high b value scheme, IVIM-derived Dfast and PF tend to demonstrate better efficacy in evaluating malignancy-related characteristics such as angiogenesis and cellular proliferation in gliomas. • Intravoxel incoherent motion-diffusion-weighted imaging (IVIM-DWI)-derived fast diffusion (Dfast) and perfusion fraction (PF) can quantitatively reflect intratumor pVEGF and pMIB-1. • IVIM-DWI-derived Dfast and PF tend to demonstrate better efficacy in evaluating glioma malignancy when an optimized scheme is used. • IVIM-DWI-derived Dfast5000 and PF5000 are promising non-invasive parameters correlating with pVEGF and pMIB-1 in gliomas.