title: Exploring Morphological and Biochemical Linkages in Fungal Growth with Label-Free Light Sheet Microscopy and Raman Spectroscopy.
abstract: Imaging tip growth in fungal hyphae is highly warranted to unravel the molecular mechanism of this extraordinarily precise and localized phenomenon. In situ probing of fungal cultures, however, have been challenging due to their inherent complexity and light penetration issues associated with conventional optical imaging. In this work, we report a label-free approach using a combination of light sheet microscopy and Raman spectroscopy to obtain concomitant morphological and biochemical information from the growing specimen. We show that the variance in morphology in the symbiotic fungus Piriformospora indica are rooted in the underlying differences in chemical composition in the specific growth zones. Our findings suggest that this potent two-pronged approach can comprehensively characterize growth areas and elucidate microbe interactions in still developing colonies with high sensitivity and multiplexing capability.
authors.affiliation: Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
authors.forename: Soumik
authors.initials: S
authors.lastname: Siddhanta
authors.affiliation: Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
authors.forename: Santosh Kumar
authors.initials: SK
authors.lastname: Paidi
authors.affiliation: Department of Plant Biology, University of Minnesota, MN, 55108, USA.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Bushley
authors.affiliation: Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.|Amity Institute of Microbial Technology, Amity University, Noida, 201303, India), Tel: (+91) 874-585-5570, Fax: (+91) 120-243-1878.
authors.forename: Ram
authors.initials: R
authors.lastname: Prasad
authors.affiliation: Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.|Department of Oncology, Johns Hopkins School of Medicine, Baltimore, MD, 21287, USA), Tel: (+1) 410-516-0656, Fax: (+1) 410-516-4316.
authors.forename: Ishan
authors.initials: I
authors.lastname: Barman
mesh_terms: D001487:Basidiomycota / Q000737:chemistry*; D025301:Hyphae / Q000254:growth & development*; D008853:Microscopy; D013059:Spectrum Analysis, Raman
keywords: label-free imaging; light sheet fluorescence microscopy; partial least squares-discriminant analysis; piriformospora indica; surface-enhanced Raman spectroscopy