Hyperspectral coherent anti-Stokes Raman scattering volumetric projection microscopy for assessing drug-cell...

Area of Science:

• Cell Biology
• Microscopy Techniques
• Pharmacology

Background:

• Lipid droplets are dynamic organelles crucial for cellular physiology and drug efficacy assessment.
• Coherent Anti-Stokes Raman Scattering (CARS) microscopy offers label-free, high-resolution imaging for drug evaluation.
• Traditional CARS microscopy's 3D imaging is slow, causes photodamage, and affects cellular microenvironments.

Purpose of the Study:

• To develop an improved CARS microscopy technique for efficient and accurate quantitative analysis of lipid droplets.
• To investigate the impact of doxorubicin hydrochloride on lipid droplet accumulation in HeLa cells.
• To enable rapid visualization of lipid droplets for drug-cell interaction research.

Main Methods:

• Utilized dual Bessel beams for CARS excitation, enhancing depth of field and resolution.
• Acquired 3D information and volumetric Raman spectra using a single 2D scan.
• Employed spectral phasor segmentation to analyze lipid droplet changes in doxorubicin-treated HeLa cells.

Main Results:

• Dual Bessel beams increased the depth of field by 5 times and resolution by 1.17 times compared to Gaussian beams.
• Demonstrated rapid quantitative visualization of lipid droplets.
• Observed reduced lipid droplet accumulation in HeLa cells treated with doxorubicin hydrochloride, correlating with drug effect.

Conclusions:

• The dual Bessel beam CARS microscopy technique significantly improves 3D imaging capabilities for lipid droplet analysis.
• This method provides a faster, less damaging alternative for drug efficacy assessment and cellular research.
• The findings support the utility of this advanced microscopy for studying drug-cell interactions and quantitative lipid droplet visualization.