Single-Molecule Tracking of Carbohydrate-Binding Modules on Cellulose Using Fluorescence Microscopy

Yu San Liu, Shi You Ding, Michael E. Himmel

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus Citations

Abstract

Single-molecule fluorescence detection is an invaluable technique for the study of molecular behavior in biological systems, both in vitro and in vivo. In this chapter, we focus on detailed protocols that utilize Total Internal Reflection Fluorescence Microscopy (TIRF-M) to visualize single molecules of carbohydrate-binding module (CBM) labeled with green fluorescent protein (GFP). The content describes step-by-step sample preparation and data acquisition, processing, and analysis. These methods can also be further used to study interactions between domains of cellulase molecules and between cellulases and cellulose.

Original languageAmerican English
Title of host publicationBiomass Conversion
Subtitle of host publicationMethods and Protocols
EditorsMichael Himmel
Pages129-140
Number of pages12
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume908
ISSN (Print)1064-3745

NREL Publication Number

  • NREL/CH-2700-52558

Keywords

  • Carbohydrate-binding module
  • Cellulose
  • Fluorophore
  • Single-molecule tracking
  • Total internal reflection fluorescence microscopy

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