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Fluorescent micrograph of Mtb detected by IF and AR in mouse and guinea pig.

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posted on 2013-02-21, 01:01 authored by Gavin J. Ryan, Donald R. Hoff, Emily R. Driver, Martin I. Voskuil, Mercedes Gonzalez-Juarrero, Randall J. Basaraba, Dean C. Crick, John S. Spencer, Anne J. Lenaerts

Mtb was detected by IF (A,D,G,J), AR (B,E,H) and combined IF-AR (C,F,I) in C57BL/6 mouse lung (A–C), GKO mouse lung (D–F) and guinea pig lung (G–I). 5 µm formalin fixed paraffin embedded sections were dewaxed, rehydrated and detected by IF. Rabbit polyclonal antibody against whole Mtb cell lysate was applied overnight at 4°C and detected using an Alexafluor 488 labeled secondary antibody (green). Slides were then stained with AR (orange/red) for 30 mins then destained. Pictures were taken using a FITC and TRITC filter at 400× magnification. (J) Confocal micrograph of Mtb detected by IF in GKO lung tissue at 1000× magnification with 1.6× digital zoom.

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