Steady-state cross-correlations for live two-colour super-resolution localization data sets.

Steady-state cross-correlations for live two-colour super-resolution localization data sets.

Nature communications (2015-06-13)

Matthew B Stone, Sarah L Veatch

ABSTRACT

Cross-correlation of super-resolution images gathered from point localizations allows for robust quantification of protein co-distributions in chemically fixed cells. Here this is extended to dynamic systems through an analysis that quantifies the steady-state cross-correlation between spectrally distinguishable probes. This methodology is used to quantify the co-distribution of several mobile membrane proteins in both vesicles and live cells, including Lyn kinase and the B-cell receptor during antigen stimulation.

MATERIALS

Product Number

Brand

Product Description

204439

Sigma-Aldrich

Sodium chloride, 99.999% trace metals basis

378860

Sigma-Aldrich

Sodium chloride, random crystals, 99.9% trace metals basis

606758

Sigma-Aldrich

Formaldehyde-¹²C solution, 20% in H₂O, 99.9 atom % ¹²C

S0817

Sigma-Aldrich

Sodium chloride solution, 0.85%

451940

Sigma-Aldrich

Sodium chloride-³⁵Cl, 99 atom % ³⁵Cl

252549

Sigma-Aldrich

Formaldehyde solution, ACS reagent, 37 wt. % in H₂O, contains 10-15% Methanol as stabilizer (to prevent polymerization)

63069

Sigma-Aldrich

Magnesium chloride solution, BioUltra, Molecular Biology, ~1 M in H₂O

54457

Sigma-Aldrich

HEPES, BioUltra, Molecular Biology, ≥99.5% (T)

15512

Sigma-Aldrich

Formaldehyde solution, meets analytical specification of USP, ≥34.5 wt. %

63036

Sigma-Aldrich

Magnesium chloride solution, BioUltra, Molecular Biology, ~0.025 M in H₂O

450006

Sigma-Aldrich

Sodium chloride, AnhydroBeads^™, −10 mesh, 99.999% trace metals basis

153745

Sigma-Aldrich

Hexanoic acid, ≥99%

M1028

Sigma-Aldrich

Magnesium chloride solution, Molecular Biology, 1.00 M±0.01 M

M8787

Sigma-Aldrich

Magnesium chloride solution, PCR Reagent, 25 mM MgCI₂solution for PCR

S6546

Sigma-Aldrich

Sodium chloride solution, 5 M

S3014

Sigma-Aldrich

Sodium chloride, Molecular Biology, DNase, RNase, and protease, none detected, ≥99% (titration)

S1679

Sigma-Aldrich

Sodium chloride, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%

H6147

Sigma-Aldrich

HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)

H3375

Sigma-Aldrich

HEPES, ≥99.5% (titration)

S6150

Sigma-Aldrich

Sodium chloride, tablet

S5886

Sigma-Aldrich

Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%

H7523

Sigma-Aldrich

HEPES, BioXtra, pH 5.0-6.5 (1 M in H₂O), ≥99.5% (titration)

S8776

Sigma-Aldrich

Sodium chloride solution, 0.9% in water, BioXtra, suitable for cell culture

S7653

Sigma-Aldrich

Sodium chloride, BioXtra, ≥99.5% (AT)

H4034

Sigma-Aldrich

HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture

F8775

Sigma-Aldrich

Formaldehyde solution, Molecular Biology, 36.5-38% in H₂O

S5150

Sigma-Aldrich

Sodium chloride solution, 5 M in H₂O, BioReagent, Molecular Biology, suitable for cell culture

00457

Sigma-Aldrich

Magnesium chloride solution, 0.1 M

47608

Sigma-Aldrich

Formaldehyde solution, Molecular Biology, BioReagent, ≥36.0% in H₂O (T)

71376

Sigma-Aldrich

Sodium chloride, BioUltra, Molecular Biology, ≥99.5% (AT)