ObjectivesThe paper focuses on the development of technology of adipose tissue optical clearing using different complex hyperosmotic optical clearing agents and tissue permeability enhancers.MethodsTo quantify optical clearing efficiency, reduced scattering coefficient was estimated from the ex vivo spatially resolved backreflectance measurements using a multi‐distant fiber optical device. Tissue morphology modification was monitored with the help of histological studies.ResultsKinetics and efficiency of the optical clearing were evaluated for ex vivo abdominal fat tissue at action of dimethyl sulfoxide, diatrizoic acid, metrizoic acid, sucrose, and fructose solutions accompanied by tissue permeability enhancers, such as various modes of fractional laser microablation and sonophoresis and their combinations. Histological studies allowed us to find agents that cause minimal morphological changes of the adipose tissue.ConclusionsMaximal optical clearing efficiency of 83.5% was observed for the samples subjected to action of the fructose‐ethanol solution during 90 min in combination with gentle modes of fractional laser microablation and sonophoresis.