Related Experiment Video
Updated: May 30, 2026

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
Published on: April 25, 2021
Study of γ-cyclodextrin host-guest complex and nanotube aggregate by fluorescence correlation spectroscopy
Amit Kumar Mandal1, Dibyendu Kumar Das, Atanu Kumar Das
1Physical Chemistry Department, Indian Association for the Cultivation of Science, Jadavpur, Kolkata, 700 032, India.
Abstract:
Fluorescence correlation spectroscopy (FCS) has been used to study the formation of large nanotube aggregates involving γ-cyclodextrin (γ-CD) and coumarin 153 (C153). It is observed that the length of a γ-CD:C153 nanotube aggregate is ∼770 nm. This is ∼480 times larger than the length of a 1:1 γ-CD:C480 complex (∼1.6 nm) and ∼950 times that of a γ-CD. This implies that 950 γ-CD units are noncovalently attached in the γ-CD:C153 aggregate. Binding constants (K(b)) of both the dyes to γ-CD were obtained from the fluctuation in fluorescence intensity. The rate of association and dissociation are obtained from the inverse of τ(off) and τ(on), respectively. The binding constant for the 1:1 γ-CD:C480 complex is ∼1000 M(-1). The burst integrated fluorescence lifetime (BIFL) histogram reveals presence of three distinct lifetime 1.8 ns (18%), 2.8 ns (69%), 3.2 ns (13%). These three lifetimes correspond to C153 present in bulk water and at the end and middle of the γ-CD:C153 nanotube aggregate, respectively. The lifetime of C480 in the 1:1 γ-CD:C480 complex is found to be 3.7 ns.

