High-resolution line-scan Brillouin microscopy for live imaging of mechanical properties during embryo development

Bevilacqua, Carlo; Gomez, Juan Manuel; Fiúza, Ulla-Maj; Chan, Chii Jou; Wang, Ling; Hambura, Sebastian; Eguren, Manuel; Ellenberg, Jan; Diz-Muñoz, Alba; Leptin, Maria; Prevedel, Robert

doi:10.1038/s41592-023-01822-1

Cited by 45 publications

(21 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We find that the mechanics of the nucleus is dominated by the cytoplasmic/nuclear interface rather than the nucleoplasm. This result coincides with recent observations made with Brillouin microscopy of cells in developing ascidian embryo, where a higher Brillouin shift was observed in perinuclear regions, indicative for a higher longitudinal modulus of the interface, than both, the nucleus and the cytoplasm[74]. Likewise, observations that the nucleus has a lower mass density and longitudinal modulus than the surrounding cytoplasm[75, 58] support our data that nucleoplasm is a soft, viscoelastic solid.…”

Section: Discussionsupporting

confidence: 93%

Measuring age-dependent viscoelastic properties of organelles, cells and organisms via Time-Shared Optical Tweezer Microrheology

Català-Castro,

Ortiz-Vásquez,

Martínez-Fernández

et al. 2023

Preprint

View full text Add to dashboard Cite

Recording the mechanical response of biological samples, the cell's interior and complex fluids in general, would enable deeper understanding of cellular differentiation, ageing and drug discovery. Here, we present a time-shared optical tweezer microrheology (TimSOM) pipeline to determine the frequency- and age-dependent viscoelastic properties of biological materials. Our approach consists in splitting a single laser beam into two near-instantaneous time-shared optical traps to carry out simultaneous force and displacement measurements with sub-nanometer and sub-picoNewton accuracy during sinusoidal perturbations. Leveraging numerical and analytical models, we find solutions to commonly encountered deviations, to build an artefact-free nanorheometer. We demonstrate the versatility of the technique by 1) measuring the phase transitions of an ageing biomolecular condensate, 2) quantifying the complex viscoelastic properties of three intracellular compartments of zebrafish progenitor cells, and, using Caenorhabditis elegans, we uncover how mutations causing nuclear envelopathies soften the cytosol of intestinal cells during organismal age. Together, our advances afford rapid phenotyping of material properties inside cells and proteins blends, opening avenues for biomedical and drug screening applications.

show abstract

Section: Discussionsupporting

confidence: 93%

Measuring age-dependent viscoelastic properties of organelles, cells and organisms via Time-Shared Optical Tweezer Microrheology

Català-Castro,

Ortiz-Vásquez,

Martínez-Fernández

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Despite its current challenges, BM holds immense potential for probing the mechanical properties of biomolecular condensates and protein aggregates. Recently, there has been significant technical progress to improve the imaging speed and reduce the light doses of BM through implementations based on stimulated Brillouin scattering (SBS), pulsed SBS, and line scanning. , Future developments in this field are expected to reduce the light dose and imaging speeds further and possibly to develop super-resolution BM techniques, though this remains challenging due to limitations arising from the acoustic wavelength. Approaches that capitalize on artificial intelligence (AI) to extract further information from the Brillouin signal could also be extremely valuable.…”

Section: Part 1: Imaging-based Techniquesmentioning

confidence: 99%

Label-Free Techniques for Probing Biomolecular Condensates

Ibrahim,

Naidu,

Miljkovic

et al. 2024

ACS Nano

View full text Add to dashboard Cite

Biomolecular condensates play important roles in a wide array of fundamental biological processes, such as cellular compartmentalization, cellular regulation, and other biochemical reactions. Since their discovery and first observations, an extensive and expansive library of tools has been developed to investigate various aspects and properties, encompassing structural and compositional information, material properties, and their evolution throughout the life cycle from formation to eventual dissolution. This Review presents an overview of the expanded set of tools and methods that researchers use to probe the properties of biomolecular condensates across diverse scales of length, concentration, stiffness, and time. In particular, we review recent years’ exciting development of label-free techniques and methodologies. We broadly organize the set of tools into 3 categories: (1) imaging-based techniques, such as transmitted-light microscopy (TLM) and Brillouin microscopy (BM), (2) force spectroscopy techniques, such as atomic force microscopy (AFM) and the optical tweezer (OT), and (3) microfluidic platforms and emerging technologies. We point out the tools’ key opportunities, challenges, and future perspectives and analyze their correlative potential as well as compatibility with other techniques. Additionally, we review emerging techniques, namely, differential dynamic microscopy (DDM) and interferometric scattering microscopy (iSCAT), that have huge potential for future applications in studying biomolecular condensates. Finally, we highlight how some of these techniques can be translated for diagnostics and therapy purposes. We hope this Review serves as a useful guide for new researchers in this field and aids in advancing the development of new biophysical tools to study biomolecular condensates.

show abstract

“…Therefore, discerning the signal from the strong background is a general challenge in Brillouin microscopy. Many innovative solutions have been proposed to improve the extinction of a Brillouin spectrometer, including an iodine absorption notch filter [60], Rubidium gas filter [61], spatial light modulation for enhanced spectrometer apodization [62], background deflection aperture masks in the spectrometer [52], and interference-based approaches involving both etalons [63] and interferometer-based setups [64,65]. Antonacci et al used a reference laser beam to destructively interfere the reflection from a specific interface in a Michelson-interferometer-style setup [64], while a division by wavefront-interference method using a prism to split the beam was employed by Lepert et al to filter both reflection and Rayleigh scatter at all planes [65].…”

Section: Division By Amplitude Interference For Brillouin Measurement...mentioning

confidence: 99%

A multi-modal microscope for integrated mapping of cellular forces and Brillouin scattering with high resolution

Meek,

Busse,

Kronenberg

et al. 2024

J. Phys. Photonics

View full text Add to dashboard Cite

Mechanical forces and stiffness play key roles in the health and development of cells and tissue, but despite the physical connection between these quantities, they cannot be monitored in parallel in most cases. Here, we introduce a fully integrated microscope that combines a method for high-resolution cell force imaging (elastic resonator interference stress microscopy, ERISM) with non-contact mapping of the elastic properties of cells (via Brillouin microscopy). In order to integrate both techniques, we had to account for the strong back reflection on the surface of the microcavity used for ERISM measurements as well as the local destruction of the cavity under illumination for Brillouin microscopy measurements. Therefore, we developed an elastic optical microcavity with minimal absorption that can perform ERISM measurements without sustaining laser damage during Brillouin microscopy. Furthermore, an unequal-arm Michelson interferometer was designed to suppress the back reflection of the laser on the ERISM microcavity surface using division by amplitude interference to reduce the reflected light and enhance the Brillouin signal. We show the utility of our integrated microscope by simultaneously mapping cellular forces and Brillouin shifts in cultures of fibroblast cells.

show abstract

High-resolution line-scan Brillouin microscopy for live imaging of mechanical properties during embryo development

Cited by 45 publications

References 39 publications

Measuring age-dependent viscoelastic properties of organelles, cells and organisms via Time-Shared Optical Tweezer Microrheology

Measuring age-dependent viscoelastic properties of organelles, cells and organisms via Time-Shared Optical Tweezer Microrheology

Label-Free Techniques for Probing Biomolecular Condensates

A multi-modal microscope for integrated mapping of cellular forces and Brillouin scattering with high resolution

Contact Info

Product

Resources

About