CB 3AA3

Lecture 5 Jan 22, 2021

Saman Sadeghi
Sadegs10@mcmaster.ca
Polarization

vertically polarized light natural/unpolarized light

Analyzing Polarized Light
 Fluorescence Anisotropy

probability of the absorption is proportional to cos2θ

θ is the angle between the exciting light and the transition dipole.
Absorption depends on interaction of oscillating electric field vector of light
with the absorption dipole moment of fluorophore
Photoselection
• polarisation (P):

• anisotropy (r):

Limiting anisotropies are +0.4 and -0.2

Electronic Transitions of Phenol
excitation polarization spectra
Fluorescence Polarization Immunoassay
Protease Assay
Combination of Fluorescence Polarization and FRET
References
• Biomolecular and Bioanalytical Techniques: Theory, Methodology and
Applications Vasudevan Ramesh (Editor) ISBN: 978-1-119-48396-0 April
2019
• Fluorescence Microscopy: From Principles to Biological Applications,
Second Edition Editor(s): Ulrich Kubitscheck May 2017 Print
ISBN:9783527338375
• Bioanalytical Chemistry, 2nd Edition Susan R. Mikkelsen, Eduardo Cortón
ISBN: 978-1-118-30254-5 March 2016
• Bajar BT, Wang ES, Zhang S, Lin MZ, Chu J. A Guide to Fluorescent Protein
FRET Pairs. Sensors (Basel). 2016;16(9):1488
• Jameson DM, Ross JA. Fluorescence polarization/anisotropy in diagnostics
and imaging. Chem Rev. 2010;110(5):2685-2708