Chemically selective imaging provides a direct approach to unraveling cellular machinery and disease mechanisms. Our long term goal is to develop highly sensitive optical microscopy that allows selective imaging of single macromolecules or single molecular assemblies in live cells without fluorophore labeling, and to apply such tools for mapping intermediate metabolites in disease processes and drug pathways. The recently developed nonlinear optical microscopy based on coherent anti-Stokes Raman scattering (CARS) allows vibrational imaging with a high signal level. However, the nonresonant background limits the sensitivity and spectral selectivity. This proposal aims to push the detection sensitivity and molecular selectivity of CARS microscopy to a new level. We propose to develop three-color CARS microscopy in which three synchronized picosecond pulse trains are used to acquire on-resonance and off-resonance CARS image simultaneously. The vibrationally resonant CARS signal is extracted from the difference between the two images. We plan to apply three-color CARS microscopy to test the hypothesis of rafts in cellular membranes. We have been able to image specific lipids in model membranes using the CARS signal from the aliphatic CD stretch vibration of a lipid molecule with fully deuterated acyl chains. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21EB004966-02
Application #
7038365
Study Section
Special Emphasis Panel (ZRG1-BECM (01))
Program Officer
Zhang, Yantian
Project Start
2005-04-01
Project End
2008-03-31
Budget Start
2006-04-01
Budget End
2008-03-31
Support Year
2
Fiscal Year
2006
Total Cost
$182,338
Indirect Cost
Name
Purdue University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
Le, Thuc T; Cheng, Ji-Xin (2009) Single-cell profiling reveals the origin of phenotypic variability in adipogenesis. PLoS One 4:e5189
Wang, Han-Wei; Fu, Yan; Huff, Terry B et al. (2009) Chasing lipids in health and diseases by coherent anti-Stokes Raman scattering microscopy. Vib Spectrosc 50:160-167
Le, Thuc T; Cheng, Ji-Xin (2009) Non-Linear Optical Imaging of Obesity-Related Health Risks: Review. J Innov Opt Health Sci 2:9-25
Le, Thuc T; Huff, Terry B; Cheng, Ji-Xin (2009) Coherent anti-Stokes Raman scattering imaging of lipids in cancer metastasis. BMC Cancer 9:42
Wang, Han-Wei; Le, Thuc T; Cheng, Ji-Xin (2008) Label-free Imaging of Arterial Cells and Extracellular Matrix Using a Multimodal CARS Microscope. Opt Commun 281:1813-1822
Huff, T B; Cheng, J-X (2007) In vivo coherent anti-Stokes Raman scattering imaging of sciatic nerve tissue. J Microsc 225:175-82
Cheng, Ji-Xin (2007) Coherent anti-Stokes Raman scattering microscopy. Appl Spectrosc 61:197-208
Kang, Eunah; Robinson, Joshua; Park, Kinam et al. (2007) Paclitaxel distribution in poly(ethylene glycol)/poly(lactide-co-glycolic acid) blends and its release visualized by coherent anti-Stokes Raman scattering microscopy. J Control Release 122:261-8
Fu, Yan; Wang, Haifeng; Huff, Terry B et al. (2007) Coherent anti-Stokes Raman scattering imaging of myelin degradation reveals a calcium-dependent pathway in lyso-PtdCho-induced demyelination. J Neurosci Res 85:2870-81
Le, Thuc T; Rehrer, Charles W; Huff, Terry B et al. (2007) Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma. Mol Imaging 6:205-11

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