As the number of CuO2 layers, n, in each unit cell of a cuprate family increases, the maximum transition temperature (Tc,max) exhibits a universal bell-shaped curve with a peak at n = 3. The microscopic mechanism of this trend remains elusive. In this study, we used advanced electron microscopy to image the atomic structure of cuprates in the Bi2Sr2Can-1CunO2n+4+δ family with 1 ≤ n ≤ 9; the evolution of the charge-transfer gap size (Δ) with n can be measured simultaneously. We determined that the n dependence of Δ follows an inverted bell-shaped curve with the minimum Δ value at n = 3. The correlation between Δ, n, and Tc,max may clarify the origin of superconductivity in cuprates.