The fixed sections were embedded in paraffin (Sigma, Saint Louis, MO, USA), and stored at 4 °C.
These sections were embedded in 5% agar and then cut into 80- to 130-μm sections using a DTK-100 microslicer (Dosaka EM Co. Ltd., Kyoto, Japan) and stained with iodine.
All sections were embedded in polyaquamount (Polysciences, Inc).
Fluorescently labelled sections were embedded in Moviol (Calbiochem, Darmstadt, Germany) and analysed under a fluorescent microscope (Axioplan2, Zeiss, Jena, Germany).
After alcohol-graded dehydratation, sections were embedded in Epon and ultrathin sections were analyzed (Jeol 100 CX II).
After a final wash in PBS buffer, sections were embedded in mounting medium and data were collected on a Zeiss LSM 510 confocal microscope (Zeiss, Oberkochen, Germany).
After a final wash in 0.05 M Tris-HCl buffer pH 7.5, sections were embedded in mounting medium and collected on a Zeiss LSM 510 confocal microscope (Zeiss, Oberkochen, Germany).
Subsequently, slides were washed three times for 5 min each with PBS and sections were embedded in Vectashield mounting medium (Vector Laboratories, Burlingame, CA), covered and sealed with nail polish around the cover glass (Corning Labware and Equipment, Corning, NY).
After alcohol-graded dehydration, sections were embedded in LR White, and ultra-thin sections were incubated with rabbit anti SR-BI/CLA-1 antibody (Novus, NB400-104), gold (12 nm particles -labeled donkey anti-rabbit IgGs (Jackson Immunoresearch) being used as secondary antibodies.
Dried sections were embedded in Permount.
Stained sections were embedded in FluorSave (Calbiochem).
