Confocal software lcs lite

SLOs (spleens and LNs) from CIA mice and synovial tissues with ELSs from RA patients were embedded in O.C.T (Sakura) and 10 µm cryo-sections were cut and fixed in ice cold acetone. The slides were rehydrated/Blocked with 2% horse serum (Jackson ImmunoResearch, 008-000-121) in PBS at room temperature in a humidified chamber, then incubated at room temperature with primary Abs for 2 hrs followed by 3x wash in PBS then incubation with secondary Abs for 1 hr. After incubation with the secondary Abs, the slides were washed in PBS 3 times, dried and mounted with Vectashield Antifade Mounting Medium (Vector Laboratories), cover-slipped, and examined with a Leica TCS SP2 AOBS confocal laser-scanning microscope. Three lasers (488, 543, and 633 nm) were used and far red emission is shown as pseudo-colour. Parameters were adjusted to scan at 1024 x 1024pixel density and 8-bit pixel depth. Emissions were recorded in three separate channels, and digital images were captured and processed with Leica Confocal Software LCS Lite. The Abs  used in this study are listed in the table 1 and their concentrations ranged between 5 and 10 g/ml.