Hemispherical photography is becoming a popular technique for gap fraction measurements to characterize biophysical parameters and solar radiation in plant canopies. One of the crucial steps in the measurement of canopy gap fraction using hemispherical photography is determining the resolution of the sampling grid. In this work, the effects of varying resolutions of sampling grids by modifying the angle widths of zenithal annuli and azimuthal sectors were evaluated for leaf area and clumping indices computations. Sensitivity analysis was performed to test these effects using artificial photographs simulating ideal canopies with varying leaf area index and aggregation levels of foliage elements. Contrasting forest types, including natural tropical cloud forest and exotic plantations, were tested as real canopies. Results indicate that leaf area and clumping indices estimates are significantly affected by the variation of sampling grids. A new approach to solve the problem of null-gap segments, obscured completely by foliage, is proposed. However, the determination of optimal combinations of zenithal annuli and azimuthal sector angular widths that suit all canopy types remains a difficult practical problem that is often overlooked. Finally, theoretically sound gap fraction and size sampling regions were demonstrated for reliable estimates of canopy biophysical parameters.