Active cell proliferation contributes to the enlargement of the nascent nucleus pulposus DOI Open Access
Rose G. Long, ChangHee Lee, Clifford J. Tabin

et al.

Developmental Dynamics, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 20, 2025

Abstract Background The notochord is an embryonic organ involved in forming and patterning the spinal column. mechanism by which transforms from a continuous rod to segmented structure excluded vertebrae residing solely as nucleus pulposus within intervertebral disc understudied. current model of notochordal segmentation suggests that swelling through formation maturation vertebrate cartilage squeezes cells vertebra. Results Analysis Collagen 10, marker for hypertrophic differentiation, well evaluation changes cell density, reveal expansion vertebral precursor occurs after has already taken place. We find bulk derived accelerated proliferation itself. In proliferation, increased at importantly contributes expand area. Conclusions Our data consistent with hypothesis enlargement before vertebra undergo hypertrophy.

Language: Английский

Macromolecular Crowding, Phase Separation, and Homeostasis in the Orchestration of Bacterial Cellular Functions DOI Creative Commons
Begoña Monterroso, William Margolin, Arnold J. Boersma

et al.

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(4), P. 1899 - 1949

Published: Feb. 8, 2024

Macromolecular crowding affects the activity of proteins and functional macromolecular complexes in all cells, including bacteria. Crowding, together with physicochemical parameters such as pH, ionic strength, energy status, influences structure cytoplasm thereby indirectly function. Notably, also promotes formation biomolecular condensates by phase separation, initially identified eukaryotic cells but more recently discovered to play key functions Bacterial require a variety mechanisms maintain homeostasis, particular environments fluctuating conditions, is emerging one mechanism. In this work, we connect homeostasis function bacterial cell compare supramolecular structures found bacteria those cells. We focus on effects separation control chromosome replication, segregation, division, discuss contribution fitness adaptation environmental stress.

Language: Английский

Citations

31
Multi-molecular hyperspectral PRM-SRS microscopy DOI Creative Commons
Wenxu Zhang, Yajuan Li, Anthony A. Fung

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Feb. 21, 2024

Abstract Lipids play crucial roles in many biological processes. Mapping spatial distributions and examining the metabolic dynamics of different lipid subtypes cells tissues are critical to better understanding their aging diseases. Commonly used imaging methods (such as mass spectrometry-based, fluorescence labeling, conventional optical imaging) can disrupt native environment cells/tissues, have limited or spectral resolution, cannot distinguish subtypes. Here we present a hyperspectral platform that integrates Penalized Reference Matching algorithm with Stimulated Raman Scattering (PRM-SRS) microscopy. Using this platform, visualize identify high density lipoprotein particles human kidney, cholesterol phosphatidylethanolamine ratio inside granule mouse hippocampus, subcellular sphingosine cardiolipin brain. Our PRM-SRS displays unique advantages enhanced chemical specificity, fast data processing distinguishing organs species.

Language: Английский

Citations

20
Genome homeostasis defects drive enlarged cells into senescence DOI Creative Commons
Sandhya Manohar, Marianna E. Estrada, Federico Uliana

et al.

Molecular Cell, Journal Year: 2023, Volume and Issue: 83(22), P. 4032 - 4046.e6

Published: Nov. 1, 2023

Cellular senescence refers to an irreversible state of cell-cycle arrest and plays important roles in aging cancer biology. Because is associated with increased cell size, we used reversible arrests combined growth rate modulation study how excessive affects proliferation. We find that enlarged cells upregulate p21, which limits progression. Cells re-enter the cycle encounter replication stress well tolerated physiologically sized but causes severe DNA damage cells, ultimately resulting mitotic failure permanent withdrawal. demonstrate fail recruit 53BP1 other non-homologous end joining (NHEJ) machinery sites robustly initiate damage-dependent p53 signaling, rendering them highly sensitive genotoxic stress. propose impaired response primes for persistent replication-acquired damage, leading division exit.

Language: Английский

Citations

38
Label-free mid-infrared photothermal live-cell imaging beyond video rate DOI Creative Commons

Genki Ishigane,

Keiichiro Toda, Miu Tamamitsu

et al.

Light Science & Applications, Journal Year: 2023, Volume and Issue: 12(1)

Published: July 19, 2023

Advancement in mid-infrared (MIR) technology has led to promising biomedical applications of MIR spectroscopy, such as liquid biopsy or breath diagnosis. On the contrary, microscopy been rarely used for live biological samples an aqueous environment due lack spatial resolution and large water absorption background. Recently, photothermal (MIP) imaging proven be applicable 2D 3D single-cell with high inherited from visible light. However, maximum measurement rate limited several frames s-1, limiting its range use. Here, we develop a significantly improved wide-field MIP quantitative phase microscope two orders-of-magnitude higher signal-to-noise ratio than previous techniques demonstrate live-cell beyond video rate. We first derive optimal system design by numerically simulating thermal conduction following effect. Then, designed homemade nanosecond optical parametric oscillator full-well-capacity image sensor. Our high-speed high-spatial-resolution great potential become new tool life science, particular analysis.

Language: Английский

Citations

32
Recent advances in label-free imaging and quantification techniques for the study of lipid droplets in cells DOI
Hyeonwoo Kim, Seungeun Oh, Seongsoo Lee

et al.

Current Opinion in Cell Biology, Journal Year: 2024, Volume and Issue: 87, P. 102342 - 102342

Published: Feb. 29, 2024

Language: Английский

Citations

13
Photoswitchable polyynes for multiplexed stimulated Raman scattering microscopy with reversible light control DOI Creative Commons

Yueli Yang,

X. Bai, Fanghao Hu

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: March 22, 2024

Abstract Optical imaging with photo-controllable probes has greatly advanced biological research. With superb chemical specificity of vibrational spectroscopy, stimulated Raman scattering (SRS) microscopy is particularly promising for super-multiplexed optical rich information. Functional SRS in response to light been recently demonstrated, but multiplexed reversible photocontrol remains unaccomplished. Here, we create a multiplexing palette photoswitchable polyynes 16 frequencies by coupling asymmetric diarylethene Carbow (Carbow-switch). Through optimization both electronic and Carbow-switch displays excellent photoswitching properties under visible control large frequency change signal enhancement. Reversible spatial-selective different organelles are demonstrated living cells. We further achieve photo-selective time-lapse organelle dynamics during oxidative stress protein phase separation. The development will open up new avenues study complex interactions cells high spatiotemporal precision capability.

Language: Английский

Citations

11
Holotomography DOI
Geon Kim, Hervé Hugonnet, Kyoohyun Kim

et al.

Nature Reviews Methods Primers, Journal Year: 2024, Volume and Issue: 4(1)

Published: July 25, 2024

Language: Английский

Citations

10
The uniformity and stability of cellular mass density in mammalian cell culture DOI Creative Commons
Xili Liu, Seungeun Oh, Marc W. Kirschner

et al.

Frontiers in Cell and Developmental Biology, Journal Year: 2022, Volume and Issue: 10

Published: Oct. 12, 2022

Cell dry mass is principally determined by the sum of biosynthesis and degradation. Measurable change in occurs on a time scale hours. By contrast, cell volume can minutes altering osmotic conditions. How changes are coupled fundamental question size control. If were proportional to during growth, would always maintain same cellular density, defined as dividing volume. The accuracy stability against perturbation this proportionality has never been stringently tested. Normalized Raman Imaging (NoRI), measure both protein lipid density directly . Using new technique , we have able investigate response pharmaceutical physiological perturbations three cultured mammalian lines. We find remarkably narrow distribution within cells, that is, significantly tighter than variability or distribution. measured independent cycle. be modulated extracellular osmolytes disruptions cytoskeleton. Yet, surprisingly resistant pharmacological synthesis degradation, suggesting there must some form feedback control homeostasis when altered. such starvation senescence induce significant shifts density. begun shed light how why remains fixed yet sensitive transitions state.

Language: Английский

Citations

31
Stimulated Raman Scattering Imaging Sheds New Light on Lipid Droplet Biology DOI Creative Commons
Hao Jia, Shuhua Yue

The Journal of Physical Chemistry B, Journal Year: 2023, Volume and Issue: 127(11), P. 2381 - 2394

Published: March 10, 2023

A lipid droplet (LD) is a dynamic organelle closely associated with cellular functions and energy homeostasis. Dysregulated LD biology underlies an increasing number of human diseases, including metabolic disease, cancer, neurodegenerative disorder. Commonly used staining analytical tools have difficulty providing the information regarding distribution composition at same time. To address this problem, stimulated Raman scattering (SRS) microscopy uses intrinsic chemical contrast biomolecules to achieve both direct visualization dynamics quantitative analysis high molecular selectivity subcellular level. Recent developments tags further enhanced sensitivity specificity SRS imaging without perturbing activity. With these advantages, has offered great promise for deciphering metabolism in single live cells. This article overviews discusses latest applications as emerging platform dissect health disease.

Language: Английский

Citations

20
Quantitative chemometric phenotyping of three-dimensional liver organoids by Raman spectral imaging DOI Creative Commons
Vernon LaLone, Aleksandra Aizenshtadt, John Goertz

et al.

Cell Reports Methods, Journal Year: 2023, Volume and Issue: 3(4), P. 100440 - 100440

Published: March 31, 2023

Confocal Raman spectral imaging (RSI) enables high-content, label-free visualization of a wide range molecules in biological specimens without sample preparation. However, reliable quantification the deconvoluted spectra is needed. Here we develop an integrated bioanalytical methodology, qRamanomics, to qualify RSI as tissue phantom calibrated tool for quantitative spatial chemotyping major classes biomolecules. Next, apply qRamanomics fixed 3D liver organoids generated from stem-cell-derived or primary hepatocytes assess specimen variation and maturity. We then demonstrate utility identifying biomolecular response signatures panel liver-altering drugs, probing drug-induced compositional changes followed by

Language: Английский

Citations

20