title: In situ detection of cellulose with carbohydrate-binding modules.
abstract: Cellulose is generally found in the context of complex plant cell wall materials and mostly in association with other glycans. Cellulose-directed carbohydrate-binding modules (CBMs) can be readily adapted to a range of methods for the in situ imaging of cellulose structures within plant cell walls or other cellulose-based materials. Protocols for the preparation and selection of plant materials, their fixation and processing for preparation of sections for CBM labeling, and fluorescence imaging procedures are described. Approaches to direct methods in which CBMs are directly coupled to fluorophores and indirect methods in which staged incubations with secondary reagents are used for the fluorescence imaging of CBM binding to materials are discussed and presented.
authors.affiliation: Faculty of Biological Sciences, Centre for Plant Sciences, University of Leeds, Leeds, United Kingdom.
authors.forename: J Paul
authors.initials: JP
authors.lastname: Knox
mesh_terms: D002473:Cell Wall / Q000737:chemistry / Q000648:ultrastructure; D002482:Cellulose / Q000032:analysis* / Q000648:ultrastructure; D005456:Fluorescent Dyes / Q000032:analysis*; D008856:Microscopy, Fluorescence / Q000379:methods; D059828:Plant Cells / Q000737:chemistry* / Q000648:ultrastructure; D010944:Plants / Q000737:chemistry* / Q000648:ultrastructure; D016610:Tissue Embedding / Q000379:methods; D016707:Tissue Fixation / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D002482:Cellulose