title: Rapid quantification of inflammation in tissue samples using perfluorocarbon emulsion and fluorine-19 nuclear magnetic resonance.
abstract: Quantification of inflammation in tissue samples can be a time-intensive bottleneck in therapeutic discovery and preclinical endeavors. We describe a versatile and rapid approach to quantitatively assay macrophage burden in intact tissue samples. Perfluorocarbon (PFC) emulsion is injected intravenously, and the emulsion droplets are effectively taken up by monocytes and macrophages. These 'in situ' labeled cells participate in inflammatory events in vivo resulting in PFC accumulation at inflammatory loci. Necropsied tissues or intact organs are subjected to conventional fluorine-19 ((19)F) NMR spectroscopy to quantify the total fluorine content per sample, proportional to the macrophage burden. We applied these methods to a rat model of experimental allergic encephalomyelitis (EAE) exhibiting extensive inflammation and demyelination in the central nervous system (CNS), particularly in the spinal cord. In a cohort of EAE rats, we used (19)F NMR to derive an inflammation index (IFI) in intact CNS tissues. Immunohistochemistry was used to confirm intracellular colocalization of the PFC droplets within CNS CD68+ cells having macrophage morphology. The IFI linearly correlated to mRNA levels of CD68 via real-time PCR analysis. This (19)F NMR approach can accelerate tissue analysis by at least an order of magnitude compared with histological approaches.
authors.affiliation: Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA. eta@cmu.edu
authors.forename: Eric T
authors.initials: ET
authors.lastname: Ahrens
authors.forename: Won-Bin
authors.initials: WB
authors.lastname: Young
authors.forename: Hongyan
authors.initials: H
authors.lastname: Xu
authors.forename: Lisa K
authors.initials: LK
authors.lastname: Pusateri
mesh_terms: D000818:Animals; D003711:Demyelinating Diseases; D004655:Emulsions; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology / Q000473:pathology*; D005260:Female; D005461:Fluorine / Q000737:chemistry; D005466:Fluorocarbons / Q000737:chemistry*; D006651:Histocytochemistry; D007249:Inflammation / Q000276:immunology / Q000473:pathology*; D016014:Linear Models; D008264:Macrophages / Q000276:immunology / Q000473:pathology; D009000:Monocytes / Q000276:immunology / Q000473:pathology; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D051381:Rats; D013116:Spinal Cord / Q000276:immunology / Q000473:pathology
chemical_list: D004655:Emulsions; D005466:Fluorocarbons; D005461:Fluorine