Analytical and Bioanalytical Chemistry

, Volume 405, Issue 26, pp 8549–8559

Automated quantitative analysis of lipid accumulation and hydrolysis in living macrophages with label-free imaging

  • Wei-Wen Chen
  • Chen-Hao Chien
  • Chiung-Lin Wang
  • Huai-Hsien Wang
  • Yuh-Lin Wang
  • Shih-Torng Ding
  • Tzong-Shyuan Lee
  • Ta-Chau Chang
Research Paper

DOI: 10.1007/s00216-013-7251-0

Cite this article as:
Chen, WW., Chien, CH., Wang, CL. et al. Anal Bioanal Chem (2013) 405: 8549. doi:10.1007/s00216-013-7251-0

Abstract

The accumulation of lipids in macrophages is a key factor that promotes the formation of atherosclerotic lesions. Several methods such as biochemical assays and neutral lipid staining have been used for the detection of lipids in cells. However, a method for real-time quantitative assessment of the lipid content in living macrophages has yet to be shown, particularly for its kinetic process with drugs, due to the lack of suitable tools for non-invasive chemical detection. Here we demonstrate label-free real-time monitoring of lipid droplets (LDs) in living macrophages by using coherent anti-Stokes Raman scattering (CARS) microscopy. In addition, we have established an automated image analysis method based on maximum entropy thresholding (MET) to quantify the cellular lipid content. The result of CARS image analysis shows a good correlation (R2 > 0.9) with the measurement of biochemical assay. Using this method, we monitored the processes of lipid accumulation and hydrolysis in macrophages. We further characterized the effect of a lipid hydrolysis inhibitor (diethylumbelliferyl phosphate, DEUP) and determined the kinetic parameters such as the inhibition constant, Ki. Our work demonstrates that the automated quantitative analysis method is useful for the studies of cellular lipid metabolism and has potential for preclinical high-throughput screening of therapeutic agents related to atherosclerosis and lipid-associated disorders.

Figure

Automated quantitative analysis for the label-free detection of lipid content in living cells

Keywords

Image-based analysis Macrophages Lipid droplet Lipid hydrolysis Drug screening Coherent anti-Stokes Raman scattering (CARS) microscopy 

Supplementary material

216_2013_7251_MOESM1_ESM.pdf (3 mb)
ESM 1(PDF 3043 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Wei-Wen Chen
    • 1
    • 2
    • 3
  • Chen-Hao Chien
    • 1
    • 4
  • Chiung-Lin Wang
    • 1
  • Huai-Hsien Wang
    • 1
    • 5
  • Yuh-Lin Wang
    • 1
    • 5
  • Shih-Torng Ding
    • 6
  • Tzong-Shyuan Lee
    • 7
  • Ta-Chau Chang
    • 1
    • 2
    • 4
  1. 1.Institute of Atomic and Molecular SciencesAcademia SinicaTaipeiTaiwan
  2. 2.Molecular Science and Technology Program, Taiwan International Graduate Program, Institute of Atomic and Molecular SciencesAcademia SinicaTaipeiTaiwan
  3. 3.Department of ChemistryNational Tsing Hua UniversityHsinchuTaiwan
  4. 4.Institute of BiophotonicsNational Yang-Ming UniversityTaipeiTaiwan
  5. 5.Department of PhysicsNational Taiwan UniversityTaipeiTaiwan
  6. 6.Department of Animal Science and Technology/Institute of BiotechnologyNational Taiwan UniversityTaipeiTaiwan
  7. 7.Department of Physiology, School of MedicineNational Yang-Ming UniversityTaipeiTaiwan

Personalised recommendations