Tissue immunofluorescence assay
Spleens
and kidneys from WT and CCR2 KO mice were frozen in Tissue-Tek® O.C.T.
embedding medium (Sakura Finetek, Torrance, CA, USA), and cut into 10
μm-thick cryosections, followed by fixation with ice-cold acetone for 5
min. Sections were incubated in 5% BSA (4240GR100, BioFroxx, Einhausen,
Germany) containing 1% anti-CD16/CD32 mAb to block Fc receptors and
then incubated with the primary Abs overnight at 4 °C. Next, sections
were incubated with Dylight 650 (84547, Thermo Fisher) or AF488 G/R IgG
(A11006, Thermo Fisher). Images were captured and analyzed using the NIS
elements AR 5.01 software (Nikon).