Figure 2. Preprocessing of 2D time-lapse stack before ROI detection.

A. Gamma adjustment enhances the contrast between the object of interest and background signal. B. Background signal correction involves selecting an ROI in the background region, measuring its average intensity, and subtracting the signal from the original image. C. The maximum intensity projection along the time axis generates a comprehensive view of the cell’s overall trajectory over the course of imaging. Defining this ROI facilitates easier detection and analysis of the cell within it across all time points.