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.   Significance:  Speckle variation induced by intracellular motion (IM) in the urothelium was observed in optical coherence tomography (OCT) images. IM can be used as a dynamic contrast to segment the urothelium by comparing two sequential OCT images. This method opens the possibility of specifically tracking the distribution of urothelial cancerous cells for identifying the microinvasion of bladder tumors.   Approach:  OCT images were acquired  ex vivo  with fresh porcine bladder tissue. IM was analyzed by tracking speckle variation using autocorrelation function, then quantified with constrained regularization method for inverting data (CONTIN method) to identify the decorrelation time (DT) of the speckle variations. Variance analysis was also conducted to show IM amplitude and distribution in the urothelium. The segmentation of the urothelium was demonstrated with OCT images with a visible urothelial layer and OCT images with an invisible urothelial layer.   Results:  Significant speckle variation induced by IM was observed in the urothelium. However, the distribution of the IM is heterogeneous. The DTs are mostly concentrated between 1 and 30 ms. With the IM as a dynamic contrast, the urothelium can be accurately and exclusively segmented, even the urothelial layer is invisible in normal OCT images.   Conclusions:  IM can be used as a dynamic contrast to exclusively track urothelial cell distribution. This contrast may provide a new mechanism for OCT to image the invasion depth and pattern of urothelial cancerous cells for accurately substaging of bladder cancer.

Segmentation of the urothelium in optical coherence tomography images with dynamic contrast