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bioRxiv · 10.1101/2023.10.08.561435

Motion Blur Microscopy

Goreke, U.·Gonzales, A.·Shipley, B.·Man, Y.·Wulftange, W.·An, R.·Hinczewski, M.·Gurkan, U. A.
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Abstract

Imaging and characterizing the dynamics of cellular adhesion in blood samples is of fundamental importance in understanding biological function. In vitro microscopy methods are widely used for this task, but typically require diluting the blood with a buffer to allow for transmission of light. However whole blood provides crucial mechanical and chemical signaling cues that influence adhesion dynamics, which means that conventional approaches lack the full physiological complexity of living microvasculature. We propose to overcome this challenge by a new in vitro imaging method which we call motion blur microscopy (MBM). By decreasing the source light intensity and increasing the integration time during imaging, flowing cells are blurred, allowing us to identify adhered cells. Combined with an automated analysis using machine learning, we can for the first time reliably image cell interactions in microfluidic channels during whole blood flow. MBM provides a low cost, easy to implement alternative to intravital microscopy, the in vivo approach for studying how the whole blood environment shapes adhesion dynamics. We demonstrate the methods reproducibility and accuracy in two example systems where understanding cell interactions, adhesion, and motility is crucial--sickle red blood cells adhering to laminin, and CAR-T cells adhering to E-selectin. We illustrate the wide range of data types that can be extracted from this approach, including distributions of cell size and eccentricity, adhesion durations, trajectories and velocities of adhered cells moving on a functionalized surface, as well as correlations among these different features at the single cell level. In all cases MBM allows for rapid collection and processing of large data sets, ranging from thousands to hundreds of thousands of individual adhesion events. The method is generalizable to study adhesion mechanisms in a variety of diseases, including cancer, blood disorders, thrombosis, inflammatory and autoimmune diseases, as well as providing rich datasets for theoretical modeling of adhesion dynamics.

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Goreke, U., Gonzales, A., Shipley, B., Man, Y., Wulftange, W., An, R., Hinczewski, M., Gurkan, U. A.. 2023-10-10. Motion Blur Microscopy. https://doi.org/10.1101/2023.10.08.561435

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