A high-precision wear particle monitoring instrument based on numerical simulation, machine vision, multi-target tracking algorithm, and microfluidic technology is designed and developed in this paper. The instrument not only performs the traditional particle number and size detection, but also proposes a particle density detection model based on motion velocity. Firstly, an on-line visual microfluidic chip for oil particles motion was specially designed. Then a mathematical model relationship between particles velocity and particles diameter, particles density, oil viscosity, oil density was established by force analysis and solving the equation, which was used to explain the working principle of the instrument. Meanwhile, a multi-target tracking algorithm incorporating the characteristics of wear particles based on YOLOv5-Deepsort was custom-developed to detect the number, size, and motion characteristics of particles under the optical field of view of the image sensor. Secondly, the oil flow field distribution and the four factors affecting the particle motion characteristics in the self-developed chamber, particle size, particle material, particle radial position, and oil velocity were analyzed by using the finite element analysis in COMSOL. Finally, a pin-disk wear experimental validation results demonstrated this approach has excellent suitability to monitor the number, size, and density of wear particles with diameters above 20μm and densities above 2000 kg/m3 simultaneously. This portable oil particle monitoring instrument provides technical support for condition monitoring, fault diagnosis, and intelligent maintenance of rotating mechanical equipment.