In this Technical Note, we introduce the Smart Lipid Droplet Assay (SLDA) and showcase its output in multiple cell types at varying confluences and across different field of view sizes. We then compare the performance of the SLDA against the fluorescence marker LipidSpot 610, before finishing with a case study where we use the SLDA to quantify how oleic acid (OA) addition changes LD dynamics in pre-adipocyte cells.