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Light scattering corrections to linear dichroism spectroscopy for liposomes in shear flow using calcein fluorescence and modified Rayleigh-Gans-Debye-Mie scattering

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Dorrington, Glen, Chmel, Nikola Paul, Norton, Stephen R., Wemyss, Alan M., Lloyd, Katherine L., Praveen Amarasinghe, Don and Rodger, Alison (2018) Light scattering corrections to linear dichroism spectroscopy for liposomes in shear flow using calcein fluorescence and modified Rayleigh-Gans-Debye-Mie scattering. Biophysical Reviews, 10 (5). pp. 1385-1399. doi:10.1007/s12551-018-0458-8 ISSN 1867-2450.

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Official URL: http://dx.doi.org/10.1007/s12551-018-0458-8

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Abstract

The interpretation of data from absorbance spectroscopy experiments of liposomes in flow systems is often complicated by the fact that there is currently no easy way to account for scattering artefacts. This has proved particularly problematic for linear dichroism (LD) spectroscopy, which may be used to determine binding modes of small molecules, peptides and proteins to liposomes if we can extract the absorbance signal from the combined absorbance/scattering experiment. Equations for a modified Rayleigh-Gans-Debye (RGD) approximation to the turbidity (scattering) LD spectrum are available in the literature though have not been implemented. This review summarises the literature and shows how it can be implemented. The implementation proceeds by first determining volume loss that occurs when a spherical liposome is subjected to flow. Calcein fluorescence can be used for this purpose since at high concentrations (> 60 mM) it has low intensity fluorescence with maxima at 525 and 563 nm whereas at low concentrations (<1 mM) the fluorescence intensity is enhanced and the band shifts to 536 nm. The scattering calculation process yields the average axis ratios of the distorted liposome ellipsoids and extent of orientation of the liposomes in flow. The scattering calculations require methods to estimate liposome integrity, volume loss, and orientation when subjected to shear stresses under flow.

Item Type: Journal Article
Subjects: Q Science > QC Physics
Divisions: Faculty of Science, Engineering and Medicine > Science > Chemistry
Library of Congress Subject Headings (LCSH): Linear dichroism, Mie scattering, Liposomes, Molecules, Peptides, Proteins
Journal or Publication Title: Biophysical Reviews
Publisher: Springer
ISSN: 1867-2450
Official Date: October 2018
Dates:
DateEvent
October 2018Published
25 September 2018Available
13 September 2018Accepted
Volume: 10
Number: 5
Page Range: pp. 1385-1399
DOI: 10.1007/s12551-018-0458-8
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access (Creative Commons)
Date of first compliant deposit: 24 October 2018
Date of first compliant Open Access: 29 October 2018
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
EP/F500378/1, MOAC-DTC [EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266

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