Table 1. Applications of FCS techniques to research of biomolecular liquid–liquid phase separation in vivo and in intro.
| Research subjects | Molecular properties | FCS techniques | |
| a Absolution concentration refers to molar concentration calculated from the average number of fluorescent molecules detected from the FCS volume element (Eq. 9). Such a concentration is determined without constructing a reference concentration curve that is typically required using the traditional method of the spectrophotometer. Thus, the FCS-determined molar concentration is also called absolute concentration. | |||
| b FLCS: fluorescence lifetime correlation spectroscopy, an interdisciplinary technique combining fluorescence lifetime fluorescence spectroscopy and fluorescence correlation spectroscopy (Ghosh et al. 2018; Machan et al. 2014). | |||
| Live-cell applications |
Centrosome (Mahen et al. 2011) |
Diffusion coefficient |
FCS, RICS, line-scanning FCS |
|
DNA transcription site (Chong et al. 2018) |
Absolute concentrationa |
FCS |
|
|
Tight junction (Beutel et al. 2019) |
Diffusion coefficient, absolute concentration, molecular brightness |
FCS, scanning FCS |
|
|
Inner centromere(Trivedi et al. 2019) |
Diffusion coefficient |
FCS |
|
|
Nucleolus (Zhu et al. 2019) |
Diffusion coefficient, absolute concentration |
FCS |
|
|
Pre-autophagosome structure (Fujioka et al. 2020) |
Diffusion coefficient |
FCS |
|
|
Stress granule, P-body (Sanders et al. 2020) |
Absolute concentration |
FCS |
|
|
Lipid droplet-lipid droplet contact site (Lyu et al. 2021) |
Diffusion coefficient |
FCS |
|
|
Photoinducible “corelets” (Bracha et al. 2018; Shimobayashi et al. 2021) |
Diffusion coefficient, absolute concentration |
FCS |
|
|
Prion-like RNA-binding proteins (Kaur et al. 2021) |
Diffusion coefficient |
FCS |
|
|
Ribosome crowding induced LLPS (Delarue et al. 2018) |
Diffusion coefficient |
FCS |
|
|
Heterochromatin (Erdel et al. 2020) |
Translational diffusion, rotational diffusion, molecular concentration, droplet size and abundance |
FCS, Pol-FCS, ICS |
|
|
In vitro
applications |
Polypeptide-DNA (Shakya and King 2018a) |
Diffusion coefficient, molecular interaction |
FCS, FCCS |
|
Histone proteins-DNA (Shakya and King 2018b) |
Diffusion coefficient |
STED-FLCSb |
|
|
Histone proteins-nucleosome (Shakya et al. 2020) |
Absolute concentration, diffusion coefficient |
FCS |
|
|
Nucleoprotien-RNA (Alshareedah et al. 2020) |
Diffusion coefficient |
FCS |
|
|
Polypeptide-ssDNA (Alshareedah et al. 2021) |
Diffusion coefficient |
FCS |
|
|
Absolute concentration, molecular brightness, diffusion coefficient |
FCS, Dual-focus FCS |
||
|
Poly(Lys)-Poly(rU),Poly(Arg)-Poly(rU) (Fisher and Elbaum-Garfinkle 2020) |
Absolute concentration, molecular interaction (dissociation constant KD) |
FCS, Dual-focus FCS |
|
|
Peptide-Poly(rU) (Kaur et al. 2021) |
Molecular interaction |
FCS |
|
|
In vitro model of stress granule formation (Guillen-Boixet et al. 2020) |
Diffusion coefficient (hydrodynamic radius) |
FCS, scanning FCS |
|
|
ySmF-peptides (Peng et al. 2020) |
Molecular diffusion (hydrodynamic radius), molecular brightness (binding stoichiometry), molecular interaction (dissociation constant KD) |
dcFCCS |
|
|
C. elegans protein LAF-1 (Wei et al. 2017) |
Molecular concentration, molecular diffusion (diffusion coefficients, viscosity, second viral coefficients, mesh size) |
Ultrafast-scanning FCS |
|
|
Microtubule-associated Tau protein (Wen et al. 2021) |
Molecular diffusion, molecular brightness |
FRET-FCS |
|
|
SARS-CoV-2 N-protein (Cubuk et al. 2021) |
Molecular dynamics |
Nanosecond FRET-FCS |
|