OtherBASIC SCIENCE INVESTIGATIONS

In Vivo Optical Imaging of Cellular Inflammatory Response in Granuloma Formation Using Fluorescence-Labeled Macrophages

Michel Eisenblätter, Jan Ehrchen, Georg Varga, Cord Sunderkötter, Walter Heindel, Johannes Roth, Christoph Bremer and Alexander Wall
Journal of Nuclear Medicine October 2009, 50 (10) 1676-1682; DOI: https://doi.org/10.2967/jnumed.108.060707

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Figures

  • FIGURE 1. 
    FIGURE 1. 

    Contrast enhancement of biogel pellets after injection of labeled Mϕs. (A) Fluorescence reflectance images were repeatedly acquired up to 144 h after injection of labeled Mϕs (5 × 106). (B) Note stronger enhancement of lipopolysaccharide (LPS) pellets (right flank, ▪), compared with control pellets without lipopolysaccharide (left flank, □). Lipopolysaccharide pellets showed increasing conspicuity over first 72 h, with slight drop of signal intensity at 144 h. Means ± SEMs are shown (n = 5). *P < 0.05, Student t test. AU = arbitrary units.

  • FIGURE 2. 
    FIGURE 2. 

    FMT of mice showed distribution of Mϕs inside pellets. (A) FMT was performed 72 h after injection of labeled Mϕs (5 × 106) to resolve 3-dimensional distribution of Mϕs in target tissue (left, NIRF image; right, color-encoded FMT). FMT confirmed FRI results and showed that Mϕs distributed mainly in periphery of pellets. (B) Fluorescence quantification by FMT showed approximately 2.7 times higher signal in lipopolysaccharide-containing pellets (▪) than in controls (□). Means ± SEMs are shown (n = 10). *P < 0.05, Student t test. AU = arbitrary units.

  • FIGURE 3. 
    FIGURE 3. 

    (A) Signal intensity of pellets after injection of different numbers of labeled Mϕs. (B) Numbers of DiR-labeled Mϕs recovered from pellets. Lipopolysaccharide-containing biogel pellets were implanted into mice, and different amounts of DiR-labeled Mϕs were injected. FMT and cell recovery were analyzed 72 h after cell injection. FMT showed clear increase of signal intensity dependant on number of injected cells (A). There was also clear dependency between amount of injected cells and numbers of DiR-positive Mϕs recovered from pellets (B). Means ± SEMs are shown (n = 5). *P < 0.05. **P < 0.01, ANOVA. AU = arbitrary units.

  • FIGURE 4. 
    FIGURE 4. 

    Migration of DiR-stained Mϕs in vivo. Mice were injected with 5 × 106 DiR-stained Mϕs. Biogel pellets were recovered 2 d after injection of Mϕs, and intensity of DiR fluorescence and surface antigen expression were measured by flow cytometry.

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