Quantitative analysis of focal adhesion dynamics using photonic resonator outcoupler microscopy (PROM)

Autor: Yue Zhuo, Ji Sun Choi, Brendan A.C. Harley, Hojeong Yu, Brian T. Cunningham, Thibault Marin
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Zdroj: Light: Science & Applications, Vol 7, Iss 1, Pp 1-15 (2018)
Light, Science & Applications
ISSN: 2047-7538
DOI: 10.1038/s41377-018-0001-5
Popis: Focal adhesions are critical cell membrane components that regulate adhesion and migration and have cluster dimensions that correlate closely with adhesion engagement and migration speed. We utilized a label-free approach for dynamic, long-term, quantitative imaging of cell–surface interactions called photonic resonator outcoupler microscopy (PROM) in which membrane-associated protein aggregates outcoupled photons from the resonant evanescent field of a photonic crystal biosensor, resulting in a highly localized reduction of the reflected light intensity. By mapping the changes in the resonant reflected peak intensity from the biosensor surface, we demonstrate the ability of PROM to detect focal adhesion dimensions. Similar spatial distributions can be observed between PROM images and fluorescence-labeled images of focal adhesion areas in dental epithelial stem cells. In particular, we demonstrate that cell–surface contacts and focal adhesion formation can be imaged by two orthogonal label-free modalities in PROM simultaneously, providing a general-purpose tool for kinetic, high axial-resolution monitoring of cell interactions with basement membranes.
Biophotonics: imaging cell adhesion A new imaging technique looks set to provide insights into how biological cells adhere and interact with surfaces. Yue Zhuo and coworkers from the University of Illinois at Urbana-Champaign in the USA have developed a technique called photonic resonator outcoupler microscopy (PROM). In the approach, cells to be studied are attached to the top surface of a photonic crystal biosensor, which is illuminated from below with focused light from a LED. The presence of cellular focal adhesions, a specialized protein located near the cell membrane that dictates how the cell interacts with external surfaces, serves to outcouple evanescent light from the biosensor resulting in a reduction in the sensor’s reflected signal. Measurement of this signal thus makes it possible to image focal adhesions and study their size and dynamic behavior.
Databáze: OpenAIRE