A 4×4 Isotropic Homogeneous Metasurface (IHM) backed, rectangular slotted dual-layered triband microstrip patch antenna (MPA) is proposed in this article. A detailed mathematical way to analyze the tensor surface impedance matrix (TSIM) of the proposed 4×4 IHM and slotted patch is demonstrated. The proposed IHM-backed MPA has a triband even-mode resonance at fr0=2.45 GHz, 2fr0=5.4 GHz, and 4fr0=7.7 GHz. A better and more accurate estimation of bio-superstrate loading effect for triband resonance is employed using the human skin (εr=52.79, σs=1.39) and blood (εr=52.8, σs=1.23) layer on the top of dual-layered metasurface-backed slotted MPA. TSIM characterization is observed for both blood- and skin layer-loaded dual-layered metasurface-backed slotted MPA. Chicken breast (εr=55.2, σs=1.49, and tanδ=0.325) with blood (εr=58.2, σs=2.59, and tanδ=0.364) is loaded onto fabricated dual-layered structure to verify numerically modeled estimation. A closed form triband, even-order resonance mode (i.e., fr0, 2fr0, and 4fr0) estimation based on dielectric superstrate thickness (Hs) is numerically established using MATLAB solver. Complete measurement setup with simulated vs. measured results is compared in this article.