Abstract
Single-particle tracking has been used extensively to advance our understanding of the plasma membrane and the mechanisms controlling the movement of cell surface proteins. These studies provide fundamental insights into the regulation of membrane receptor activation and the assembly of signaling clusters. Here, we describe a method to label and track B cell receptor (BCR) and other cell surface proteins and how this method can be adapted to simultaneously track two molecular species or examine the movement of membrane proteins in relation to membrane microdomains. We recently used this method to study the role of the actin cytoskeleton in the regulation of B cell receptor dynamics at the cell surface.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Saxton MJ, Jacobson K (1997) Single-particle tracking: applications to membrane dynamics. Annu Rev Biophys Biomol Struct 26:373–399
Andrews NL, Lidke KA, Pfeiffer JR, Burns AR, Wilson BS, Oliver JM, Lidke DS (2008) Actin restricts FcepsilonRI diffusion and facilitates antigen-induced receptor immobilization. Nat Cell Biol 10:955–963
Espenel C, Margeat E, Dosset P, Arduise C, le Grimellec C, Royer CA, Boucheix C, Rubinstein E, Milhiet PE (2008) Single-molecule analysis of CD9 dynamics and partitioning reveals multiple modes of interaction in the tetraspanin web. J Cell Biol 182:765–776
Fujiwara T, Ritchie K, Murakoshi H, Jacobson K, Kusumi A (2002) Phospholipids undergo hop diffusion in compartmentalized cell membrane. J Cell Biol 157:1071–1081
Edidin M, Zuniga MC, Sheetz MP (1994) Truncation mutants define and locate cytoplasmic barriers to lateral mobility of membrane glycoproteins. Proc Natl Acad Sci U S A 91:3378–3382
Jaqaman K, Kuwata H, Touret N, Collins R, Trimble WS, Danuser G, Grinstein S (2011) Cytoskeletal control of CD36 diffusion promotes its receptor and signaling function. Cell 146:593–606
Treanor B, Depoil D, Gonzalez-Granja A, Barral P, Weber M, Dushek O, Bruckbauer A, Batista FD (2010) The membrane skeleton controls diffusion dynamics and signaling through the B cell receptor. Immunity 32:187–199
Crocker JC, Grier DG (1996) Methods of digital video microscopy for colloidal studies. J Colloid Interface Sci 179:298–310
Ritchie K, Shan XY, Kondo J, Iwasawa K, Fujiwara T, Kusumi A (2005) Detection of non-Brownian diffusion in the cell membrane in single molecule tracking. Biophys J 88:2266–2277
Otsu N (1979) A threshold selection method from gray-level histograms. IEEE Trans Sys Man Cyber 9:62–66
Liu W, Meckel T, Tolar P, Sohn HW, Pierce SK (2010) Intrinsic properties of immunoglobulin IgG1 isotype-switched B cell receptors promote microclustering and the initiation of signaling. Immunity 32:778–789
Tolar P, Hanna J, Krueger PD, Pierce SK (2009) The constant region of the membrane immunoglobulin mediates B cell-receptor clustering and signaling in response to membrane antigens. Immunity 30:44–55
Fleire SJ, Batista FD (2009) Studying cell-to-cell interactions: an easy method of tethering ligands on artificial membranes. Methods Mol Biol 462:145–154
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Wasim, L., Treanor, B. (2018). Single-Particle Tracking of Cell Surface Proteins. In: Liu, C. (eds) B Cell Receptor Signaling. Methods in Molecular Biology, vol 1707. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7474-0_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7474-0_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7473-3
Online ISBN: 978-1-4939-7474-0
eBook Packages: Springer Protocols