Fitc conjugated anti gfp

Spleens were embedded in OCT media (Sakura Finetek) and flash frozen in liquid nitrogen chilled isopentane. Sections were cut at a thickness of 5 um and mounted on glass slides. Sections were re-hydrated at room temperature in PBS for 10 minutes and then blocked at room temperature with PBS containing 3% BSA for 20 minutes. FITC conjugated anti-GFP (Rockland Immunochemicals) was used to enhance the signal from YFP in infected cells. Cellular markers were detected with Alexa 647conjugated anti-IgD (BD Biosciences) and purified anti-FDC-M1 (BD Biosciences) followed by Alexa 546 conjugated anti-rat IgG (Invitrogen). All images were collected on an Axiovert 200M fluorescent microscope (Carl Zeiss) using Axiovision version 4.7 imaging software.

Spleens from infected mice were embedded in OCT media (Sakura Finetek) and flash frozen in chilled isopentane (Fisher Scientific). Frozen sections were cut at a thickness of 5 um and stored at −80°C until needed. To detect infected cells, FITC conjugated anti-GFP (Rockland Immunochemicals) was used to enhance the signal of H2bYFP. IgD was detected using an Alexa 647 conjugated anti-IgD (BD Biosciences). Follicular dendritic cells were detected using a purified anti-FDC-M1 (BD Biosciences followed by Alexa 546 conjugated anti-rat (Invitrogen). All images were collected on an Axiovert 200M fluorescent microscope (Carl Zeiss) using Axiovision version 4.7 imaging software.