Uncovering solvation heterogeneity in live cells via hyperspectral stimulated Raman scattering microscopy

2021 ◽  
Author(s):  
Xiaoqi Lang ◽  
Kevin Welsher
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Live Cells ◽  
Stimulated Raman

scholarly journals Utilizing Stimulated Raman Scattering Microscopy To Study Intracellular Distribution of Label-Free Ponatinib in Live Cells

2019 ◽  
Vol 63 (5) ◽  
pp. 2028-2034 ◽  
Author(s):  
Kristel Sepp ◽  
Martin Lee ◽  
Marie T. J. Bluntzer ◽  
G. Vignir Helgason ◽  
Alison N. Hulme ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Intracellular Distribution ◽  
Live Cells ◽  
Label Free ◽  
Stimulated Raman

scholarly journals Vibrational Imaging of Glucose Uptake in Live Cells and Tissues by Stimulated Raman Scattering Microscopy

2015 ◽  
Vol 108 (2) ◽  
pp. 480a
Author(s):  
Fanghao Hu ◽  
Zhixing Chen ◽  
Luyuan Zhang ◽  
Yihui Shen ◽  
Lu Wei ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Glucose Uptake ◽  
Stimulated Raman Scattering ◽  
Live Cells ◽  
Stimulated Raman

scholarly journals Mapping Solvation Heterogeneity in Live Cells by Hyperspectral Stimulated Raman Scattering Microscopy

2020 ◽  
Author(s):  
Xiaoqi Lang ◽  
Kevin Welsher
Keyword(s):  
Raman Scattering ◽  
Near Infrared ◽  
Stimulated Raman Scattering ◽  
Live Cells ◽  
Spatiotemporal Resolution ◽  
Environmental Variance ◽  
Dynamic Matrix ◽  
Coherent Raman ◽  
Living Organisms ◽  
Stimulated Raman

<p>Water provides a dynamic matrix in which all biochemical processes occur in living organisms. The structure and dynamics of intracellular water constitute the cornerstone for understanding all aspects of cellular function. Fundamentally, direct visualization of subcellular solvation heterogeneity is essential but remains challenging with commonly used NMR methods due to poor spatial resolution. To explore this question, we demonstrate a vibrational-shift imaging approach by combining the spectral-focusing hyperspectral stimulated Raman scattering (hsSRS) technique with an environmentally-sensitive nitrile probe. The sensing ability of a near-infrared nitrile-containing molecule is validated in the solution phase, microscopic droplets and cellular environments. Finally, we quantitatively measure the subcellular solvation variance between the cytoplasm (29.5%, S.E. 1.8%) and the nucleus (57.3%, S.E. 1.0%), which is in good agreement with previous studies. This work sheds light on heterogenous solvation in live systems using coherent Raman microscopy and opens up new avenues to explore environmental variance in complex systems with high spatiotemporal resolution.</p>

Mapping Solvation Heterogeneity in Live Cells by Hyperspectral Stimulated Raman Scattering Microscopy

2019 ◽  
Author(s):  
Xiaoqi Lang ◽  
Kevin Welsher
Keyword(s):  
Raman Scattering ◽  
Near Infrared ◽  
Stimulated Raman Scattering ◽  
Free Water ◽  
Live Cells ◽  
Spatiotemporal Resolution ◽  
Environmental Variance ◽  
Dynamic Matrix ◽  
Coherent Raman ◽  
Stimulated Raman

<div><div><div><p>Water provides a dynamic matrix in which all biochemical processes occur in living organisms. The structure and dynamics of intracellular water constitute the cornerstone for understanding all aspects of cellular function. Fundamentally, direct visualization of subcellular solvation heterogeneity is essential but remains challenging with commonly used NMR methods due to poor spatial resolution. To explore this question, we demonstrate a vibrational-shift imaging approach by combining the spectral-focusing hyperspectral stimulated Raman scattering (hsSRS) technique with an environmentally-sensitive nitrile probe. The sensing ability of a near-infrared nitrile-containing molecule is validated in the solution phase, microscopic droplets and cellular environments. Finally, we quantitatively measure the subcellular solvation variance in the degree of free water content between the cytoplasm (29.7%, S.E. 1.1%) and the nucleus (56.4%, S.E. 1.3%), which is in good agreement with previous studies. This work sheds light on mapping the heterogenous solvation in live systems using coherent Raman microscopy and opens up new avenues to explore environmental variance in complex systems with high spatiotemporal resolution.</p></div></div></div>

scholarly journals Vibrational Imaging of Glucose Uptake Activity in Live Cells and Tissues by Stimulated Raman Scattering

2015 ◽  
Vol 54 (34) ◽  
pp. 9821-9825 ◽  
Author(s):  
Fanghao Hu ◽  
Zhixing Chen ◽  
Luyuan Zhang ◽  
Yihui Shen ◽  
Lu Wei ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Glucose Uptake ◽  
Stimulated Raman Scattering ◽  
Live Cells ◽  
Uptake Activity ◽  
Stimulated Raman

scholarly journals Live-Cell Imaging and Quantification of PolyQ Aggregates by Stimulated Raman Scattering of Selective Deuterium Labeling

2019 ◽  
Author(s):  
Kun Miao ◽  
Lu Wei
Keyword(s):  
Raman Scattering ◽  
Live Cell Imaging ◽  
Neurodegenerative Disorder ◽  
Stimulated Raman Scattering ◽  
Live Cells ◽  
Deuterium Labeling ◽  
Multicellular Organisms ◽  
Subcellular Resolution ◽  
Stimulated Raman

AbstractHuntington’s disease, a major neurodegenerative disorder, involves deposition of aggregation-prone proteins with long polyglutamine (polyQ) expansions. The ability to non-perturbatively visualize the formation of aggregates could offer new molecular insight for their pathologic roles. Here, we propose stimulated Raman scattering imaging of deuterium-labeled glutamine to investigate native polyQ aggregates in live cells with subcellular resolution. Through the enrichment of deuterated glutamine in the polyQ sequence of mutant Huntingtin (mHtt) proteins, we first achieved sensitive and specific SRS imaging of carbon-deuterium bonds (C-D) from aggregates without GFP labeling. These aggregates become 1.8-fold denser compared to those with GFP. Second, we performed ratiometric quantification, which revealed a dependence of protein compositions on aggregation sizes. Moreover, we calculated the absolute concentrations for sequestered mHtt and non-mHtt proteins within the same aggregates. Our method may readily reveal new features of polyQ aggregates and could be suited for in vivo investigations on multicellular organisms.

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