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Abstract
Cellulose is composed of tightly packed glucan chains that give it superior strength but render it recalcitrant to breakdown. The presence of dislocations susceptible to degradation within cellulose fibrils has been postulated for decades, but they have never been directly visualized. We have developed a super-resolution fluorescence microscopy approach to visualize alternating crystalline and disordered regions within bacterial cellulose fibrils. The measured size of crystalline regions ranges from 40 – 400 nm and matches the length distribution of cellulose nanocrystals produced through acid hydrolysis, while disordered regions are 20 – 120 nm long, heterogeneous and do not form large amorphous pockets. These results lend strong support for the fringed-micellar model of cellulose fibrils and provide fundamental insight into the role of cellulose nanostructure in its breakdown by chemical and enzymatic means.
Supplementary materials
Title
Materials and Methods, seven supplementary figures and one table
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Figure S1. Labeling BC with different reaction schemes and with spectrally distinct fluorophores.
Figure S2. Continuous localization of fluorophores along Cy5-BC fibrils and identification of bright regions using a local median threshold method.
Figure S3. Different thresholding methods used for quantitatively characterizing the labeling pattern of DTAF-BC and Cy5-BC fibrils.
Figure S4. Visualizing the cellulose labeling pattern using different super-resolution imaging techniques.
Figure S5. AFM images of BC and CNCs produced following various durations of the acid hydrolysis treatment.
Figure S6. Lengths of CNCs produced from bacterial cellulose with different durations of the acid hydrolysis treatment compared to Cy5_5μM-BC spacing lengths measured with different peak-picking thresholds.
Figure S7. Proposed mechanism for the acid hydrolysis of bacterial cellulose fibrils in the production of cellulose nanocrystals.
Table S1. Characterization of purified BC and bacterial CNCs resulting from time-lapsed acid hydrolysis.
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Data for manuscript "Direct super-resolution imaging of the disordered and crystalline nanostructure of bacterial cellulose fibrils"
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All raw data used in this manuscript is included in this repository.
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