Temporal Domain Multiplexing (TDM)

TEMPORAL DOMAIN MULTIPLEXING Simulation of temporal domain multiplexing using two far-red fluorescent proteins. A 21-frame line plot derived from a HeLa cell Region of Interest (ROI); the plot illustrates the distinct photobleaching kinetics of mfRFP and mfRFP-A, which together constitute the cumulative signal in the raw image. CODE FOLDER: TDM Unmixing MATLAB code files and the reference photobleaching curve of individual signal (mfRFP and mfRFP-A) present in the raw image. DATA FOLDER: Acquired brief movie (Time series image) of HeLa cell expressing mfRFP and mfRFP-A fusion tags in H2B and cytoplasmic fusion tags. Images were acquired using 594nmSoRA excitation laser channel. RESULT FOLDER: The signal unmixing result (MATLAB) elucidates the signal contribution of each FPS (OUTPUT) to their respective corresponding structures H2B and cytoplasm. Note that the image colors used above red-H2B and green-cytoplasm are pseudo colors to enhance visualization and both H2B and the cytoplasm were indeed red.