Macropinocytosis is the process of cell drinking in which plasma membrane ruffles extend, curve, and return to the plasma membrane surface to form large intracellular organelles called macropinosomes. This complex process can be carefully understood through the constant advancements in multidimensional fluorescence microscopy, specifically lattice light sheet (LLS), however, there follows a demand for data visualization tools capable of displaying the complex cellular processes. Although LLS microscopes produce stunning movies of cellular activity with high spatial and temporal resolution, data presentation is a significant barrier for researchers that are new to multidimensional microscopy. Experiments using LLS microscopes require understanding fluorescence microscopy, cell biology, data management, computer science, and most importantly presentation of 5 dimensions of data (x, y, z, t, λ) into a scientifically accurate 2-dimensional manuscript. The first objective addresses how to develop an experiment for LLS microscopy, since many traditional experiments require unique preparations and considerations. Experimental development is followed by a discussion of the limitations and applications of the different visualization tools that were developed including volumetric intensities, orthogonal planes, isosurfaces, and mesh overlays. Furthermore, a non-programmingbased step-by-step protocol is presented using ChimeraX for LLS data visualization. These techniques are applied to understand macropinocytosis, a process where actin-rich plasma membrane ruffles form into large intracellular organelles, in a phosphatidylinositol 3- kinase (PI3K)-coordinated manner. The visualization tools show that macropinosomes form through multiple ruffling morphologies, PI3K activity varies throughout the process, and that 3’-phosphoinositides play a critical role in macropinosome sealing but not ruffling. Through the development and application of LLS visualization techniques, these data indicate that local PI3K activity is amplified in ruffles and serves as a priming mechanism for the closure and sealing of ruffles into macropinosomes.