Shedding new light on lipid functions with CARS and SRS microscopy

Yu, Yong Ming; Ramachandran, Prasanna Venkatesh; Wang, Meng C.

doi:10.1016/j.bbalip.2014.02.003

Cited by 71 publications

(66 citation statements)

References 66 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this section, we will summarize the applications of vibrational tags in studying metabolisms of cholesterol, fatty acids and choline. For more general reviews about applications of Raman imaging in lipid biology, the readers could refer to earlier reviews by Yu et al 31 and Syed et al 32 …”

Section: Lipid Metabolismmentioning

confidence: 99%

Applications of vibrational tags in biological imaging by Raman microscopy

Zhao

Shen

et al. 2017

Analyst

View full text Add to dashboard Cite

As a superb tool to visualize and study the spatial-temporal distribution of chemicals, Raman microscopy has made big impacts to many disciplines of science. While the label-free imaging has been the prevailing strategy in Raman microscopy, recent development and applications of vibrational/Raman tags, particularly when coupled with stimulated Raman Scattering (SRS) microscopy, have generated intense excitement in biomedical imaging. SRS imaging of vibrational tags has enabled researchers to study a wide range of small biomolecules with high specificity, sensitivity and multiplex capability, at single live cell level, tissue level or even in vivo. As reviewed in the article, this platform has facilitated imaging distribution and dynamics of small molecules such as glucose, lipids, amino acids, nucleic acids, and drugs that are otherwise difficult to do with other means. As both the vibrational tags and Raman instrumental development progress rapidly and synergistically, we anticipate that the technique will shed light onto an even broader spectrum of biomedical problems.

show abstract

Section: Lipid Metabolismmentioning

confidence: 99%

Applications of vibrational tags in biological imaging by Raman microscopy

Zhao

Shen

et al. 2017

Analyst

View full text Add to dashboard Cite

show abstract

“…without perturbation. Label-free chemical and biochemical investigations (5,6) transduce the desired signal without an exogenous label (fluorescent, radioactive, or otherwise) representing an essential step toward this goal. Many label-free methods require one of the interacting species to be either tethered or immobilized to the sensor surface, introducing a potential perturbation to the natural state of the species (7,8).…”

mentioning

confidence: 99%

Origin and prediction of free-solution interaction studies performed label-free

Bornhop

Kammer

Kussrow

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Interaction/reaction assays have led to significant scientific discoveries in the biochemical, medical, and chemical disciplines. Several fundamental driving forces form the basis of intermolecular and intramolecular interactions in chemical and biochemical systems (London dispersion, hydrogen bonding, hydrophobic, and electrostatic), and in the past three decades the sophistication and power of techniques to interrogate these processes has developed at an unprecedented rate. In particular, label-free methods have flourished, such as NMR, mass spectrometry (MS), surface plasmon resonance (SPR), biolayer interferometry (BLI), and backscattering interferometry (BSI), which can facilitate assays without altering the participating components. The shortcoming of most refractive index (RI)-based label-free methods such as BLI and SPR is the requirement to tether one of the interaction entities to a sensor surface. This is not the case for BSI. Here, our hypothesis is that the signal origin for freesolution, label-free determinations can be attributed to conformation and hydration-induced changes in the solution RI. We propose a model for the free-solution response function (FreeSRF) and show that, when quality bound and unbound structural data are available, FreeSRF correlates well with the experiment (R 2 > 0.99, Spearman rank correlation coefficients >0.9) and the model is predictive within ∼15% of the experimental binding signal. It is also demonstrated that a simple mass-weighted dη/dC response function is the incorrect equation to determine that the change in RI is produced by binding or folding event in free solution.backscattering interferometry | assay methodology | molecular interactions | conformation change | hydration change C ontemporary assays enabling single-molecule detection (1,2) have accelerated the sequencing of the human genome (3) and facilitated imaging with extraordinary resolution without labels (4). To most closely approximate the natural state, an interaction assay methodology would interrogate the processes (reaction, molecular interaction, protein folding event, etc.) without perturbation. Label-free chemical and biochemical investigations (5, 6) transduce the desired signal without an exogenous label (fluorescent, radioactive, or otherwise) representing an essential step toward this goal. Many label-free methods require one of the interacting species to be either tethered or immobilized to the sensor surface, introducing a potential perturbation to the natural state of the species (7,8). However, back-scattering interferometry (BSI) is a free-solution label-free technique with the added benefit of sensitivity that rivals fluorescence (9). There are other techniques performed in free solution, such as MS (10, 11) and NMR (12,13) and the widely used isothermal titration calorimetry (ITC) (14, 15). As with NMR, ITC has many advantages, but exhibits modest sensitivity and often requires large sample quantities. Another increasingly popular free-solution approach is microscale thermophoresis (...

show abstract

“…Comparative analysis of label and label-free methods is available in (Yen et al, 2010; Yu et al, 2014). Biochemical quantification assays such as TLC or GC/LC-MS can provide detailed analysis of the composition of various lipid molecules.…”

Section: Support Protocol 2: Preparation Of Frozen Tissue Sections Fomentioning

confidence: 99%

“…Comprehensive descriptions of the underlying physical principles are available elsewhere for interested researchers (Min et al, 2011). Detailed information on the various components of an SRS microscope setup is also available for further reference (Freudiger et al, 2008; Saar et al, 2010; Yu et al, 2014). …”

Section: Introductionmentioning

confidence: 99%

Label‐Free Biomedical Imaging of Lipids by Stimulated Raman Scattering Microscopy

Ramachandran

Mutlu

Wang

2015

CP Molecular Biology

Self Cite

View full text Add to dashboard Cite

Advances in modern optical microscopy have provided unparalleled access to intracellular structure and function, yet visualizing lipid molecules within a cell remains challenging. Stimulated Raman Scattering (SRS) microscopy is a recently developed imaging modality that addresses this challenge. By selectively imaging the vibration of chemical moieties enriched in lipids, this technique allows for rapid imaging of lipid molecules in vivo without the need for perturbative extrinsic labels. SRS microscopy has been effectively employed in the study of fat metabolism, helping uncover novel regulators of lipid storage. This unit provides a brief introduction to the principle of SRS microscopy, and describes methods for its use in imaging lipids in cells, tissues and whole organisms.

show abstract

Shedding new light on lipid functions with CARS and SRS microscopy

Cited by 71 publications

References 66 publications

Applications of vibrational tags in biological imaging by Raman microscopy

Applications of vibrational tags in biological imaging by Raman microscopy

Origin and prediction of free-solution interaction studies performed label-free

Label‐Free Biomedical Imaging of Lipids by Stimulated Raman Scattering Microscopy

Contact Info

Product

Resources

About