Cy5 labeled goat anti rabbit antibody

Immunofluorescence and confocal microscopy were performed on frozen sections using: rat anti-mouse F4/80 (Serotec Ltd, Oxford, UK); Cy3-labeled mouse anti-mouse α-SMA (Sigma, St. Louis, MO); goat anti-collagen I (Southern Tech, Birmingham, AL, USA); FITC-labeled rat anti-mouse CD206 (Serotec Ltd, Oxford, UK); rabbit anti-CX3CR1 (Prosci incorporated, Poway, CA); Cy5-labeled goat anti-rabbit antibody (Invitrogen, Carlsbad, CA, USA) and Alexa555-labeled donkey anti-goat antibody (Invitrogen, Carlsbad, CA). Sections were examined using a fluorescent microscope (Model: Axioplan2 imaging, Carl Zeiss, Oberkoche, Germany) or confocal microscope (Model: LSM 510 META, Carl Zeiss). In addition, a single MMT cell co-expressing F4/80 and α-SMA antigens was examined by a Z-stack image. Single, double, or triple positive cells were counted in 10 high-power fields (× 40) per section by means of a 0.0625 mm² graticule fitted in the eyepiece of the microscope and expressed as cells per mm².

Immunofluorescence and confocal microscopy were performed on frozen sections. Macrophages were labeled with rat anti-mouse F4/80 (Serotec Ltd, Oxford, UK) or FITC-conjugated mouse anti-human CD68 (DAKO, Carpinteria, CA, USA), while myofibroblasts were detected with the Cy3-labeled mouse anti-mouse/human α-SMA (Sigma, St. Louis, MO, USA). Other antibodies used include goat anti-collagen I (Southern Tech, Birmingham, AL, USA), FITC-labeled rat anti-mouse CD206 (Serotec Ltd), Cy5-labeled goat anti-rabbit antibody (Invitrogen, Carlsbad, CA, USA) and Alexa555-labeled donkey anti-goat antibody (Invitrogen). Positive signals were examined using a fluorescent microscope (Axioplan2 imaging; Carl Zeiss, Oberkoche, Germany) or a confocal microscope (LSM 510 META, Carl Zeiss) and single-, double- or triple-positive cells were counted in 10 high-power fields ( × 40) per section and expressed as cells per square millimeter.