Introduction
Tissue engineering develops materials that mimic biological tissues [1]. Often the basis for these materials are cells which are cultured on scaffolds [2]. The engineered tissues can be used to repair or even replace the body’s own material.
Mollie Smoak’s research focuses on the synthesis, processing, and evaluation of new biomaterials for use as scaffolds to support the regeneration of musculoskeletal tissues [2]. Specifically, these materials recapitulate the biochemical cues, physicochemical stimuli, and native architecture found in these tissues. Moreover, these biomaterials could be utilized as carriers for controlled drug delivery, non-viral vectors for gene therapy, and platforms for modeling disease [1,2].
Challenges
Depending on their thickness, sharp imaging of the utilized material and scaffolds can be a challenge for optical widefield microscopy. For this study, fibers were visualized with conventional widefield imaging, but, unfortunately, there is a lot of unwanted background signal which makes image interpretation difficult.
Methods
The scaffold developed for this experimental work is composed of electrospun fibers which were labeled with fluorescein. The goal was to study the fiber morphology and pore characteristics of the scaffold. The fibers were imaged with a THUNDER Imager 3D Cell Culture, an inverted microscope platform equipped with opto-digital LVCC (Large Volume Computational Clearing) technology [3]. It enables users to obtain a clear view of details, even deep within a whole, intact sample, in real time without the out-of-focus blur typical of conventional widefield systems.
Results
With a THUNDER Imager using LVCC, the final processed image is much sharper compared to the original raw image. The individual fibers of the scaffold can be clearly resolved and then analyzed more precisely.
References
- A. Atala, R. Lanza, A.G. Mikos, R. Nerem, Eds., Principles of Regenerative Medicine, 3rd Edition (Academic Press, Elsevier, 2019) ISBN 978-0-12-809880-6, DOI: 10.1016/C2015-0-02433-9.
- M. Smoak, A.G. Mikos, Advances in biomaterials for skeletal muscle engineering and obstacles still to overcome, Materials Today Bio (2020) vol. 7, 100069, DOI: 10.1016/j.mtbio.2020.100069.
- J. Schumacher, L. Bertrand, THUNDER Technology Note: THUNDER Imagers: How Do They Really Work?, Science Lab (2019) Leica Microsystems.
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