Laser Raman Tweezer Spectroscopy (RTS) for the Molecular and Functional Characterization of Single Live Mouse Mammary Tumor Initiating Cells (TIC)
Abstract
The main goal of this proposal is to assess whether Raman Spectroscopy (RS) combined with optical tweezer technology (RTS) can be utilized in order to identify distinct subpopulations of highly tumorigenic cells among an otherwise homogeneous population of tumor initiating cells derived from a p53 null mouse mammary tumor model. Laser activated Raman backscattered light from cells generates spectral profiles (wavelengths). The spectral profiles represent molecular and biochemical signatures or intracellular molecular profiles (IMPs) of living cells. Our working hypothesis is that tumor-initiating cells (lineage- CD24hi/CD29hi) derived from p53null mouse mammary tumors contain distinct subpopulation of cells with various tumorigenic potential. RTS, by intracellular molecular profiling, would allow one to distinguish heterogeneity among an otherwise homogeneous population of tumor initiating cells (TICs). Methods: Individual TICs were sorted by flow cytometry, placed on a quartz slide (high thermal conductivity) and radiated for RS measurements. Laser beam power was optimized to maximize Raman signature and minimize cell damage. Significant Findings: RS identified distinct subpopulations within the TICs derived from individual p53 null mouse mammary tumors. We have demonstrated that Raman Spectroscopy (which identifies cells with unique intracellular molecular profiles) combined with flow cytometry (which identifies cells with unique extracellular surface molecules) may be utilized in order to distinguish heterogeneity among an otherwise homogeneous population of TICs. RS induced minimal damage to the cells and there was minimal interference on RS measurement by the quartz slide. Single live tumor initiating cells generated profiles in the wavelength of 600-1800 watts. Future aims are to isolate cells derived from the distinct subpopulations within the TICs and to assess their in vitro and in vivo tumorigenic potential.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 2013
- Accession Number
- ADA601816
Entities
People
- Charles Ye
- Daniel Neuman
- David Moore
- Fariba Behbod
- Hanan Elsarraj
- Yan Hong
Organizations
- University of Kansas Medical Center