Correlative all-optical quantification of mass density and mechanics of sub-cellular compartments with fluorescence specificity

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

Abstract

Quantitative measurements of physical parameters become increasingly important for understanding biological processes. Brillouin microscopy (BM) has recently emerged as one technique providing the 3D distribution of viscoelastic properties inside biological samples — so far relying on the implicit assumption that refractive index (RI) and density can be neglected. Here, we present a novel method (FOB microscopy) combining BM with optical diffraction tomography and epi-fluorescence imaging for explicitly measuring the Brillouin shift, RI and absolute density with specificity to fluorescently labeled structures. We show that neglecting the RI and density might lead to erroneous conclusions. Investigating the nucleoplasm of wild-type HeLa cells, we find that it has lower density but higher longitudinal modulus than the cytoplasm. Thus, the longitudinal modulus is not merely sensitive to the water content of the sample — a postulate vividly discussed in the field. We demonstrate the further utility of FOB on various biological systems including adipocytes and intracellular membraneless compartments. FOB microscopy can provide unexpected scientific discoveries and shed quantitative light on processes such as phase separation and transition inside living cells.

Details

OriginalspracheEnglisch
Aufsatznummere68490
Seitenumfang23
FachzeitschrifteLife
Jahrgang11
PublikationsstatusVeröffentlicht - 10 Jan. 2022
Peer-Review-StatusJa

Externe IDs

PubMed 35001870
WOS 000758043100001
ORCID /0000-0003-4017-6505/work/142253805

Schlagworte

Forschungsprofillinien der TU Dresden

DFG-Fachsystematik nach Fachkollegium

Schlagwörter

  • Brillouin microscopy, HeLa cells, Human, density measurement, mechanical properties, optical diffraction tomography, phase transition, Density measurement, Optical diffraction tomography, Mechanical properties, Phase transition, Cells/cytology, Humans, Tomography, Optical/methods, Fluorescence, Intracellular Space, Microscopy/methods, Refractometry, HeLa Cells, Cell Nucleus