The code analyzes photobleaching from a timeseries image TIF image (single image file with multiple frames). Such a TIF file can be generated from a sequence of image files using standard software (i.e., ImageJ).
The photobleaching code can process a tiff image stack and detect bright objects of arbitrary size and shape. It can calculate the brightness/intensity of those objects in each frame and produce a intensity vs time plot. The code can be used to analyze photobleaching rate.
- Can handle multiple objects in a frame
- Can handle irregularly shaped objects
- Can dynamically adjust the mask every frame to account for object drift
- Average birghtness of all the detected beads in the frames are calculated
- OpenCV
- Matplotlib
- NumPy
- tkinter/ttkbootstrap
The following figure shows the average fluorescence intensity vs time plot. A time series image set containing multiple beads is used as the input data.
This work was developed at the Hesselink research lab, Stanford University. The work was partially supported by the National Institute of Health (NIH) Grant R01GM138716 and 5R21HG009758.