11 May 2018 Polyethylene glycol molecular weight influences the ClearT2 optical clearing method for spheroids imaging by confocal laser scanning microscopy
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Abstract
Some fluorescence microscopy techniques, such as confocal laser scanning microscopy (CLSM), have a limited penetration depth. Consequently, the visualization and imaging of three-dimensional (3-D) cell cultures, such as spheroids, using these methods can be a significant challenge. Therefore, to improve the imaging of 3-D tissues, optical clearing methods have been optimized to render transparency to the opaque spheroids. The influence of the polyethylene glycol (PEG) molecular weight (MW) used in the ClearT2 method for the imaging of propidium iodide (PI)-stained spheroids was investigated. The results demonstrated that the ClearT2 clearing method contributes to spheroids transparency and to the preservation of PI fluorescence intensity for all the PEG MW used (4000, 8000, and 10,000 Da). Furthermore, the ClearT2 method performed using PEG 4000 Da allowed a better PI signal penetration depth and cross-section depth. Overall, the optimization of PEG MW can improve the imaging of intact spheroids by CLSM. Furthermore, this work may also contribute to increase the application of 3-D cell culture models by the pharmaceutical industry for the high-throughput screening of therapeutics.
© 2018 Society of Photo-Optical Instrumentation Engineers (SPIE)
Elisa C. Costa, Elisa C. Costa, André F. Moreira, André F. Moreira, Duarte de Melo-Diogo, Duarte de Melo-Diogo, Ilidio J. Correia, Ilidio J. Correia, } "Polyethylene glycol molecular weight influences the ClearT2 optical clearing method for spheroids imaging by confocal laser scanning microscopy," Journal of Biomedical Optics 23(5), 055003 (11 May 2018). https://doi.org/10.1117/1.JBO.23.5.055003 . Submission: Received: 3 February 2018; Accepted: 17 April 2018
Received: 3 February 2018; Accepted: 17 April 2018; Published: 11 May 2018
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