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Biomedical applications of two-foci cross-correlation technique in massively parallel fluorescence correlation spectroscopy

  • Stanko N. Nikolic*
  • , Sho Oasa
  • , Aleksandar J. Krmpot
  • , Lars Terenius
  • , Milivoj R. Belić
  • , Vladana Vukojević
  • *Corresponding author for this work
  • University of Belgrade
  • Karolinska Institutet

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

Abstract

We report the use of two-foci cross-correlation in a massively parallel fluorescence correlation spectroscopy (mpFCS) setup to map the spatial distribution and motion of fluorescent molecules inside living cells. The mpFCS experimental setup enables the simultaneous measurement of raw photon counts from a matrix of confocal observation volume elements (OVEs) generated in parallel. To determine the average number of fluorescent molecules in each OVE, their diffusion times, and the two-dimensional velocity components in the imaging plane, we compute autocorrelation functions (ACFs) and two-foci cross-correlation functions (tfCCFs) after each data acquisition. We demonstrate the power of this approach by measuring, in live HEK 293 cells expressing the Glucocorticoid Receptor tagged at the Nterminus with Enhanced Green Fluorescent Protein (eGFP-GR), the directionality of nucleocytoplasmic transport.

Original languageEnglish
Title of host publicationDynamics and Fluctuations in Biomedical Photonics XXIII
EditorsVV Tuchin, MJ Leahy, RK Wang
PublisherSPIE
Number of pages6
Volume13850
ISBN (Electronic)978-1-5106-9614-3
ISBN (Print)978-1-5106-9613-6
DOIs
Publication statusPublished - 5 Mar 2026
Event23rd Dynamics and Fluctuations in Biomedical Photonics - San Francisco, United States
Duration: 17 Jan 202619 Jan 2026

Publication series

NameProgress In Biomedical Optics And Imaging

Conference

Conference23rd Dynamics and Fluctuations in Biomedical Photonics
Country/TerritoryUnited States
CitySan Francisco
Period17/01/2619/01/26

Keywords

  • Fluorescence Correlation Spectroscopy
  • Nucleocytoplasmic Transport of Glucocorticoid Receptor

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