摘要： Real-time tracking of multiple particles is key for quantitative analysis of dynamic biophysical processes and materials science via time-lapse microscopy image data, especially for single molecule biophysical techniques, such as magnetic tweezers and centrifugal force microscopy. However, real-time multiple particle tracking with high resolution is limited by the current imaging processes or tracking algorithms. Here, we demonstrate 1 nm resolution in three dimensions in real-time with a graphics-processing unit (GPU) based on a compute unified device architecture (CUDA) parallel computing framework instead of only a central processing unit (CPU). We also explore the trade-offs between processing speed and size of the utilized regions of interest and a maximum speedup of 137 is achieved with the GPU compared with the CPU. Moreover, we utilize this method with our recently self-built centrifugal force microscope (CFM) in experiments that track multiple DNA-tethered particles. Our approach paves the way for high-throughput single molecule techniques with high resolution and efficiency.