Label-Free Visualization and Quantification of Biochemical Markers of Atherosclerotic Plaque Progression Using Intravascular Fluorescence Lifetime.
Julien Bec, Deborah Vela, Jennifer E Phipps, Michael Agung, Jakob Unger, Kenneth B Margulies, Jeffrey A Southard, L Maximilian Buja, Laura Marcu
Author Information
Julien Bec: Department of Biomedical Engineering, University of California, Davis, California, USA.
Deborah Vela: Department of Cardiovascular Pathology Research, Texas Heart Institute, Houston, Texas, USA.
Jennifer E Phipps: Department of Biomedical Engineering, University of California, Davis, California, USA.
Michael Agung: Department of Biomedical Engineering, University of California, Davis, California, USA.
Jakob Unger: Department of Biomedical Engineering, University of California, Davis, California, USA.
Kenneth B Margulies: Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Jeffrey A Southard: Division of Cardiovascular Medicine, UC Davis Health System, University of California-Davis, Sacramento, California, USA.
L Maximilian Buja: Department of Cardiovascular Pathology Research, Texas Heart Institute, Houston, Texas, USA; Department of Pathology and Laboratory Medicine, the University of Texas Health Science Center at Houston, Houston, Texas, USA.
Laura Marcu: Department of Biomedical Engineering, University of California, Davis, California, USA. Electronic address: lmarcu@ucdavis.edu.
OBJECTIVES: This study aimed to systematically investigate whether plaque autofluorescence properties assessed with intravascular fluorescence lifetime imaging (FLIm) can provide qualitative and quantitative information about intimal composition and improve the characterization of atherosclerosis lesions. BACKGROUND: Despite advances in cardiovascular diagnostics, the analytic tools and imaging technologies currently available have limited capabilities for evaluating in situ biochemical changes associated with luminal surface features. Earlier studies of small number of samples have shown differences among the autofluorescence lifetime signature of well-defined lesions, but a systematic pixel-level evaluation of fluorescence signatures associated with various histological features is lacking and needed to better understand the origins of fluorescence contrast. METHODS: Human coronary artery segments (n = 32) were analyzed with a bimodal catheter system combining multispectral FLIm with intravascular ultrasonography compatible with in vivo coronary imaging. Various histological components present along the luminal surface (200-μm depth) were systematically tabulated (12 sectors) from each serial histological section (n = 204). Morphological information provided by ultrasonography allowed for the accurate registration of imaging data with histology data. The relationships between histological findings and FLIm parameters obtained from 3 spectral channels at each measurement location (n = 33,980) were characterized. RESULTS: Our findings indicate that fluorescence lifetime from different spectral bands can be used to quantitatively predict the superficial presence of macrophage foam cells (mFCs) (area under the receiver-operator characteristic curve: 0.94) and extracellular lipid content in advanced lesions (lifetime increase in 540-nm band), detect superficial calcium (lifetime decrease in 450-nm band area under the receiver-operator characteristic curve: 0.90), and possibly detect lesions consistent with active plaque formation such as pathological intimal thickening and healed thrombus regions (lifetime increase in 390-nm band). CONCLUSIONS: Our findings indicate that autofluorescence lifetime provides valuable information for characterizing atherosclerotic lesions in coronary arteries. Specifically, FLIm can be used to identify key phenomena linked with plaque progression (e.g., peroxidized-lipid-rich mFC accumulation and recent plaque formation).
