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Imaging the Morphological Structure of Silk Fibroin Constructs through Fluorescence Energy Transfer and Confocal Microscopy

1
CNR Nanotec, Institute for Nanotechnology, National Council of Research, Via Monteroni, 73100 Lecce, Italy
2
Department of Industrial Engineering, University of Trento, Via Sommarive 9, 38123 Trento, Italy
3
BIOtech Research Center, Department of Industrial Engineering, University of Trento, Via delle Regole 18, 38123 Trento, Italy
*
Author to whom correspondence should be addressed.
Academic Editor: Seokheun Choi
Electron. Mater. 2021, 2(2), 186-197; https://doi.org/10.3390/electronicmat2020015
Received: 30 March 2021 / Revised: 20 May 2021 / Accepted: 27 May 2021 / Published: 3 June 2021
(This article belongs to the Special Issue Feature Papers of Electronic Materials)
Silk fibroin is a well-known biopolymer that is used in several applications in which interactions with biological tissue are required. Fibroin is extremely versatile and can be shaped to form several constructs that are useful in tissue engineering applications. Confocal imaging is usually performed to test cell behavior on a construct, and, in this context, the fibroin intrinsic fluorescence is regarded as a problem. In addition, the intrinsic fluorescence is not intense enough to provide useful morphological images. In fact, to study the construct’s morphology, other techniques are used (i.e., SEM and Micro-CT). In this work, we propose a method based on fluorescence energy transfer (FRET) to suppress the fibroin intrinsic fluorescence and move it to a higher wavelength that is accessible to confocal microscopy for direct imaging. This was done by creating two FRET couples by dispersing two fluorophores (2,5-diphenyloxazole (PPO) and Lumogen F Violet 570 (LV)) into the fibroin matrix and optimizing their percentages to suppress the fibroin intrinsic fluorescence. With the optimized composition, we produced an electrospun mat, and the dimensions of the fibers were accurately determined by confocal microscopy. View Full-Text
Keywords: silk fibroin; Fluorescence Resonance Energy Transfer (FRET); confocal imaging; electrospinning silk fibroin; Fluorescence Resonance Energy Transfer (FRET); confocal imaging; electrospinning
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MDPI and ACS Style

Bucciarelli, A.; Quaranta, A.; Maniglio, D. Imaging the Morphological Structure of Silk Fibroin Constructs through Fluorescence Energy Transfer and Confocal Microscopy. Electron. Mater. 2021, 2, 186-197. https://doi.org/10.3390/electronicmat2020015

AMA Style

Bucciarelli A, Quaranta A, Maniglio D. Imaging the Morphological Structure of Silk Fibroin Constructs through Fluorescence Energy Transfer and Confocal Microscopy. Electronic Materials. 2021; 2(2):186-197. https://doi.org/10.3390/electronicmat2020015

Chicago/Turabian Style

Bucciarelli, Alessio, Alberto Quaranta, and Devid Maniglio. 2021. "Imaging the Morphological Structure of Silk Fibroin Constructs through Fluorescence Energy Transfer and Confocal Microscopy" Electronic Materials 2, no. 2: 186-197. https://doi.org/10.3390/electronicmat2020015

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