This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We have developed a pallet array nanotechnology that permits isolation of individual adherent cells. Combining this nanotechnology with antibody-based detection methods, advanced optical imaging and fine needle aspirate (FNA) sampling of breast tumors, we are piloting a methodology that 1) overcomes limitations to existing technologies, such as laser microdissection, 2) could be available to patients at the time of diagnosis vs. after tumor resection, 3) permits enumeration of various cellular elements present within a tumor potentially yielding important information for prognosis or predictive of therapeutic efficacy, 4) is designed for high throughput automated analyses, and 4) has the potential to assess the molecular profile of individual cellular compartments, which could provide additional information for the design of tailored individualized therapy. The HYPOTHESIS for these studies is that the pallet array nanotechnology will permit the identification, enumeration, and isolation of the following individual cellular elements from primary breast tumors: breast cancer stem cells, endothelial progenitor cells, myoepithelial cells, and inflammatory infiltrate. This hypothesis will be tested by pursuing the following Specific Aims: 1. Establish the detection threshold for identifying rare adherent cells. We will employ mixtures of cells expressing unique combinations of cell surface molecules in varying proportions, confirmed by flow cytometry, applied to the pallet array. As detection of multiple tumor cellular subsets will require multicolor/multi-antigen detection, we will perform multi-color fluorescence imaging to establish the detection threshold for rare cells. Additionally, we will refine the design of the pallet array. 2. Apply this methodology to primary breast cancer cells using Fine Needle Aspirate (FNA) sampling. Pilot feasibility studies will be conducted on breast tumor specimens using FNAs performed immediately after resection, in order to test the ability to generalize the findings from cell lines to primary tumor tissues. These studies will establish isolation, identification, and detection methods for the enumeration of breast cancer stem cells, endothelial progenitor cells, myoepithelial cells, tumor epithelial cells, and inflammatory infiltrates.

National Institute of Health (NIH)
National Center for Research Resources (NCRR)
Biotechnology Resource Grants (P41)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-SBIB-L (40))
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of California Irvine
Schools of Medicine
United States
Zip Code
Malacrida, Leonel; Astrada, Soledad; Briva, Arturo et al. (2016) Spectral phasor analysis of LAURDAN fluorescence in live A549 lung cells to study the hydration and time evolution of intracellular lamellar body-like structures. Biochim Biophys Acta 1858:2625-2635
Choi, Bernard; Tan, Wenbin; Jia, Wangcun et al. (2016) The Role of Laser Speckle Imaging in Port-Wine Stain Research: Recent Advances and Opportunities. IEEE J Sel Top Quantum Electron 2016:
Alfonso-García, Alba; Pfisterer, Simon G; Riezman, Howard et al. (2016) D38-cholesterol as a Raman active probe for imaging intracellular cholesterol storage. J Biomed Opt 21:61003
Alfonso-García, Alba; Smith, Tim D; Datta, Rupsa et al. (2016) Label-free identification of macrophage phenotype by fluorescence lifetime imaging microscopy. J Biomed Opt 21:46005
Libby, Andrew E; Wang, Hong; Mittal, Richa et al. (2015) Lipoprotein lipase is an important modulator of lipid uptake and storage in hypothalamic neurons. Biochem Biophys Res Commun 465:287-92
Datta, Rupsa; Alfonso-García, Alba; Cinco, Rachel et al. (2015) Fluorescence lifetime imaging of endogenous biomarker of oxidative stress. Sci Rep 5:9848
Badran, Karam W; Manuel, Cyrus T; Loy, Anthony Chin et al. (2015) Long-term in vivo electromechanical reshaping for auricular reconstruction in the New Zealand white rabbit model. Laryngoscope 125:2058-66
Chlebicki, Cara A; Protsenko, Dmitry E; Wong, Brian J (2014) Preliminary investigations on therapy thresholds for laser dosimetry, cryogen spray cooling duration, and treatment cycles for laser cartilage reshaping in the New Zealand white rabbit auricle. Lasers Med Sci 29:1099-109
Rohde, S B; Kim, A D (2014) Convolution model of the diffuse reflectance for layered tissues. Opt Lett 39:154-7
Rohrbach, Daniel J; Muffoletto, Daniel; Huihui, Jonathan et al. (2014) Preoperative mapping of nonmelanoma skin cancer using spatial frequency domain and ultrasound imaging. Acad Radiol 21:263-70

Showing the most recent 10 out of 648 publications