Burst Analysis Spectroscopy is a fluorescence technique developed in my lab to explore and resolve complex populations of macromolecules in free-solution. Together with my collaborators at TAMU in the lab of Hays Rye, BAS has been used to investigate a variety of dynamic, non-equilibrium processes that are at the core of a wide range of biological processes (e.g. protein aggregation and dis-aggregation; vesicle assembly and disassembly; phage DNA loading). We have recently expanded the method to include multi-color particles (mcBAS).

Publications:

• Jason Puchalla, Kelly Krantz, Robert Austin, and Hays Rye. Burst analysis spectroscopy: A versatile single-particle approach for studying distributions of protein aggregates and fluorescent assemblies. Proceedings of the National Academy of Sciences Sep 2008, 105 (38) 14400-14405

• Krantz, KC, Puchalla, J, Thapa, R, Kobayashi, C, Bisher, M, Viehweg, J et al.. Clathrin coat disassembly by the yeast Hsc70/Ssa1p and auxilin/Swa2p proteins observed by single-particle burst analysis spectroscopy. J. Biol. Chem. 2013;288 (37):26721-30.

• Lin, Z, Puchalla, J, Shoup, D, Rye, HS. Repetitive protein unfolding by the trans ring of the GroEL-GroES chaperonin complex stimulates folding. J. Biol. Chem. 2013;288 (43):30944-55.

• Brooks, A, Shoup, D, Kustigian, L, Puchalla, J, Carr, CM, Rye, HS et al.. Single particle fluorescence burst analysis of epsin induced membrane fission. PLoS ONE. 2015;10 (3):e0119563.

• Weaver, J, Jiang, M, Roth, A, Puchalla, J, Zhang, J, Rye, HS et al.. GroEL actively stimulates folding of the endogenous substrate protein PepQ. Nat Commun. 2017;8 :15934.

• Shoup, D , Roth, A, Thapa, R, Puchalla, J, Rye, H. Development and application of multicolor burst analysis spectroscopy. Biophys J. 2021 Jun 1;120(11):2192-2204. doi: 10.1016/j.bpj.2021.03.035. Epub 2021 Apr 5.

Useful links:

• Prep of multicolor fluorescent beads: http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.607.3630&rep=rep1&type=pdf

• Commercial beads (0.1um and larger; 4 mixed colors) https://www.thermofisher.com/order/catalog/product/T7279#/T7279