Bilayer graphene has drawn significant attention due to the opening of a band gap in its low energy electronic spectrum, which offers a promising route to electronic applications. The gap can be either tunable through an external electric field or spontaneously formed through an interaction-induced symmetry breaking. Our scanning tunneling measurements reveal the microscopic nature of the bilayer gap to be very different from what is observed in previous macroscopic measurements or expected from current theoretical models. The potential difference between the layers, which is proportional to charge imbalance and determines the gap value, shows strong dependence on the disorder potential, varying spatially in both magnitude and sign on a microscopic level. Furthermore, the gap does not vanish at small charge densities. Additional interaction-induced effects are observed in a magnetic field with the opening of a subgap when the zero orbital Landau level is placed at the Fermi energy.Bilayer graphene consists of two graphene sheets overlaid in the Bernal stacking orientation where A 2 atoms of the top layer lie on top of the B 1 atoms of the bottom layer (see * These authors contributed equally to this work.§ To whom correspondence should be addressed: nikolai.zhitenev@nist.gov, joseph.stroscio@nist.gov.2 Fig. 1a), connected by the interlayer coupling 1 γ , thus breaking the A/B sublattice symmetry in the individual graphene layers. This results in massive chiral fermions where the electronic energy dispersion is hyperbolic in momentum, in contrast to the linear dispersion that leads to massless carriers in single layer graphene 1,2 . In bilayer graphene the energy bands still meet at the charge neutrality point (E D ) in the absence of an electric field between the layers (neglecting interaction effects) (Fig. 1b). In an applied electric field a potential asymmetry is developed between the layers, resulting in the opening of an energy band gap between the low lying bands making bilayer graphene of intense interest in electronic applications ( Fig. 1b) [2][3][4][5][6][7][8][9][10] . Bilayer graphene also differs from single layer graphene in its magnetic quantization in the quantum Hall regime. At E D , the four-fold degenerate Landau level (LL) in single layer graphene becomes eight-fold degenerate in the bilayer due to the additional layer degeneracy 3,11,12 . When the gap is opened this manifold splits into two four-fold degenerate quartets polarized on each layer at low energies. Lifting of these degeneracies have been observed in recent measurements [13][14][15][16] .Theoretical studies 17,18 suggest the existence of interaction-driven band gaps, which are even possible in zero applied field with corresponding quantum Hall ferromagnetic states 17,19 .The energy band gap in bilayer graphene has been studied by optical measurements such as angle resolved photoemission spectroscopy 20 and infrared spectroscopy [4][5][6]21 , which demonstrate that the gap is externally tunable and can reach values up to ≈ 250 meV...