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. 2018 Mar;188(3):672–682. doi: 10.1016/j.ajpath.2017.11.005

Figure 1.

Figure 1

Histologic characterization and localization of Borrelia burgdorferi within doxycycline and untreated rhesus macaques with late disseminated Lyme disease. A:B. burgdorferi polyclonal positive control immunofluorescence assay (IFA); rabbit B. burgdorferi hyperimmunized polyclonal (primary) and goat anti-rabbit IgG Alexa Fluor 488 (secondary) antibodies. Green fluorescence of a morphologically intact spirochete. B:B. burgdorferi monoclonal positive control IFA; mouse outer surface protein A monoclonal (primary) and goat anti-mouse IgG Alexa Fluor 568 (secondary) antibodies. Red fluorescence of morphologically intact spirochetes. C: Green and red confocal channels merged, with yellow indicating colocalization of monoclonal and polyclonal antibodies from positive control tissue. D: Cerebrum (IH11 doxycycline treated); leptomeninges are focally expanded by minimal localized lymphoplasmacytic inflammation. E: Brainstem (IH22 doxycycline treated); minimal localized lymphoplasmacytic perivascular cuffing adjacent to the fourth ventricle. F: Green fluorescence illustrates six variably sized morphologically intact spirochetes within the cerebral neuropil. G: Green fluorescence reveals two morphologically intact spirochetes within the brainstem neuropil; differential interference contrast (DIC) applied to enhance contrast. H: Ulnar nerve (IK66 doxycycline treated); minimal localized lymphoplasmacytic perivascular cuffing adjacent to the epineurium. I: Sural nerve (IL75 untreated); mild localized lymphoplasmacytic perivascular cuffing within the epineurium. J: Ulnar nerve (IP55 untreated); moderate focally extensive lymphoplasmacytic inflammation radiating outward from a vessel and dissecting neighboring epineurium. K: Green fluorescence illustrates a morphologically intact spirochete within the right tibial epineurium (IP55 untreated). L: Localized green fluorescence adjacent to left ulnar axonal nerve fibers (IP67 untreated); DIC image applied to enhance contrast. Three-dimensional imaging in inset shows the cylindrical shape with convex end of the structure, indicating that this may be a cross section at the end/pole of the spirochete; 1 unit = 1.88 μm. Scale bars: 20 μm (A, B, F, and K); 10 μm (C, G, and L). Original magnification: ×10 (D, E, and H–J); ×63 (additional ×2 amplification applied with Leica LAS X software when appropriate; A–C, F, G, K and L).