A reflectance volume holographic imaging (VHI) endoscope has been designed for simultaneous in vivo imaging of surface and subsurface tissue structures. Prior utilization of VHI systems has been limited to ex vivo tissue imaging. The VHI system presented in this work is designed for laparoscopic use. It consists of a probe section that relays light from the tissue sample to a handheld unit that contains the VHI microscope. The probe section is constructed from gradient index (GRIN) lenses that form a 1: 1 relay for image collection. The probe has an outer diameter of 3.8 mm and is capable of achieving 228.1 lp/mm resolution with 660-nm Kohler illumination. The handheld optical section operates with a magnification of 13.9 and a field of view of 390 mu m x 244 mu m. System performance is assessed through imaging of 1951 USAF resolution targets and soft tissue samples. The system has also passed sterilization procedures required for surgical use and has been used in two laparoscopic surgical procedures. (C) 2017 Society of Photo-Optical Instrumentation Engineers (SPIE)