How can you differentiate between sample movement (drift) and movement of the particle of interest?

webinar Webinar: Advancing single molecule microscopy with active stabilization

What can I do to differentiate between the whole sample moving during imaging vs the actual movement of the protein I am studying? Esp in a situation where both cases might happen in parallel?

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V. Kaye Thomas Answered Sep 19, 2022

University of Rochester Medical Center

Hello,

Use of a fiducial marker would help you to differentiate between these two movements. A good fiducial marker is basically anything that can be visualized by your system and is completely immobilized on the sample substrate. The immobile object can then be used as a reference object as the image is acquired. The fiducial marker will remain fixed within the sample, but will move if the sample drifts. Images can be normalized/aligned to the fiducial and any remaining movement attributed to the actual movement of the cell. This alignment of the fiducial can be achieved during imaging with a stage that monitors and responds to the fiducial position (as in the webinar) or in post-processing image analysis using image alignment software functions. I hope this answered your question.

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