Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview01:13

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview

1.1K
Attenuated total reflectance (ATR) infrared spectroscopy is a powerful analytical technique used to study the composition of materials. It is widely employed in chemistry, materials science, forensic science, and other fields where sample characterization is required. ATR has several advantages over traditional transmission IR spectroscopy, including the requirement of little to no sample preparation and the ability to analyze a wide range of samples.
The ATR process begins by directing a beam...
1.1K
Raman Spectroscopy Instrumentation: Overview01:26

Raman Spectroscopy Instrumentation: Overview

965
A conventional Raman spectrophotometer includes a laser source, a sample holding system, a wavelength selector, and a detector.
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
965
Calibration Curves: Correlation Coefficient01:10

Calibration Curves: Correlation Coefficient

4.4K
In a linear calibration curve, there is a value called the calibration coefficient, denoted by 'r,' which measures the strength and the direction of association between two variables. The correlation coefficient value ranges from −1 to +1. A value of +1 indicates a perfect positive linear correlation, −1 denotes a perfect negative correlation, and 0 implies no correlation between the two variables. A positive correlation value establishes that as one variable increases, the...
4.4K
Raman Spectroscopy: Overview01:20

Raman Spectroscopy: Overview

1.3K
The underlying principle of Raman spectroscopy is based on the interaction between light and matter, specifically molecules' inelastic scattering of photons. When a monochromatic beam of light, typically from a laser source, interacts with a sample, most scattered light has the same frequency as the incident light. This is known as Rayleigh scattering.
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and...
1.3K
IR Spectrometers01:25

IR Spectrometers

2.2K
There are two main infrared (IR) spectrophotometers: dispersive IR spectrometers and Fourier transform infrared (FTIR) spectrometers. In a dispersive IR spectrometer, a beam of infrared radiation produced by a hot wire is divided into two parallel equal-intensity beams using mirrors. One beam passes through the sample, while another is a reference beam. The beams then move through the monochromator, which separates the radiations into a continuous spectrum of different frequencies. The...
2.2K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Cord-blood black carbon particle burden is associated with a C19MC small extracellular vesicle miRNA signature enriched for neurodevelopmental pathways.

Environmental research·2026
Same author

Cord-blood black carbon burden is associated with coordinated inflammatory and heme-metabolic transcriptional programs at birth in Bradford, United Kingdom.

The Science of the total environment·2026
Same author

Correction to "Biocompatible Label-Free Detection of Carbon Black Particles by Femtosecond Pulsed Laser Spectroscopy".

Nano letters·2026
Same author

Black carbon is detectable in association with small extracellular vesicles in fetal circulation.

Environment international·2026
Same author

ABEL-FRET bridges the timescale gap in single-molecule measurements of the structural dynamics in the A<sub>2A</sub> adenosine receptor.

Communications chemistry·2026
Same author

Nanoscale Activity Mapping of Chloride-Permeable Pentameric Receptors.

ACS sensors·2026
Same journal

Tau protein differentially affects Piezo1 and Kir2.1 channels in brain capillary endothelial cells.

Biophysical journal·2026
Same journal

Emergent Intercellular Junction Stability during Cyclic Tissue Loading.

Biophysical journal·2026
Same journal

Enhanced-Sampling Simulations Reveal Distinct Intermediates in SARS-CoV-2 FSE Pseudoknot Interconversion.

Biophysical journal·2026
Same journal

Structure-based simulations of the full Flock House virus capsid reveal pathways and energetics of an infection-critical peptide externalization event.

Biophysical journal·2026
Same journal

Quantifying the Peripheral Surface Information Entropy from Conformational Ensembles of Globular Protein-Peptide Complexes.

Biophysical journal·2026
Same journal

Anisotropic unbinding and location-dependent hovering of a kinesin motor head over microtubule.

Biophysical journal·2026
See all related articles

Related Experiment Video

Updated: Jan 4, 2026

Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
09:46

Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging

Published on: April 28, 2022

4.6K

Raster Image Correlation Spectroscopy Performance Evaluation.

Marco Longfils1, Nick Smisdom2, Marcel Ameloot3

  • 1Department of Mathematical Sciences, Chalmers University of Technology and University of Gothenburg, Gothenburg, Sweden.

Biophysical Journal
|November 1, 2019
PubMed
Summary
This summary is machine-generated.

Raster image correlation spectroscopy (RICS) analysis provides insights into molecular mobility and concentration. This study introduces theoretical expressions to optimize RICS imaging parameters for improved accuracy in diffusion coefficient recovery.

More Related Videos

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy
09:57

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy

Published on: July 25, 2022

4.5K
Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy
06:51

Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy

Published on: August 2, 2018

7.5K

Related Experiment Videos

Last Updated: Jan 4, 2026

Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
09:46

Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging

Published on: April 28, 2022

4.6K
Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy
09:57

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy

Published on: July 25, 2022

4.5K
Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy
06:51

Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy

Published on: August 2, 2018

7.5K

Area of Science:

  • Biophysics
  • Chemical Physics
  • Optical Microscopy

Background:

  • Raster image correlation spectroscopy (RICS) is a powerful fluorescence imaging technique for analyzing molecular dynamics in cells.
  • Existing RICS protocols lack a rigorous theoretical framework for optimizing experimental parameters and predicting data quality.
  • Accurate determination of molecular mobility, concentration, and stoichiometry is crucial for understanding biological processes.

Purpose of the Study:

  • To develop a theoretical approach for predicting RICS accuracy and precision based on experimental parameters.
  • To provide explicit expressions detailing the dependence of RICS results on imaging settings and sample properties.
  • To establish practical guidelines and software for optimizing RICS data acquisition and analysis.

Main Methods:

  • Derivation of explicit theoretical expressions relating RICS parameters to experimental settings.
  • Simulation of parameter effects on diffusion coefficient recovery.
  • Development of a modified RICS calculation to correct for region-of-interest size bias.
  • Implementation of a bootstrap-based confidence interval for improved parameter estimation.
  • In vitro validation using enhanced green fluorescent protein (eGFP) in varying buffer viscosities.

Main Results:

  • Decreasing pixel size significantly reduces error in diffusion constant (D) determination.
  • Optimal scan speed dramatically improves accuracy, with up to a 25-fold reduction in mean-squared relative error for D = 1 μm²/s.
  • RICS data quality is robust across nM-μM concentrations and requires fewer images at higher molecular brightness.
  • The bootstrap confidence interval for D shows superior coverage probability compared to least-squares methods.

Conclusions:

  • Theoretical framework enables prediction and optimization of RICS experimental parameters for enhanced accuracy.
  • Practical guidelines and simulation software are provided to facilitate robust RICS data acquisition and analysis.
  • This work significantly advances the application of RICS for quantitative molecular analysis in biological systems.