Mayer s hematoxylin and eosin y

Human and mouse placenta samples were fixed with 4% paraformaldehyde in PBS for 24 hours and then embedded in paraffin. For histology, 5-mm sections were stained with Mayer’s Hematoxylin and Eosin Y solutions (Sigma-Aldrich). Immunohistochemistry was done as described previously (37 (link)). The following primary antibodies were used: anti-LC3 (1:200; Novus, NB600-1384), anti-P62 (1:500; Santa Cruz Biotechnology, SC28359), and anti-cytokeratin (1:1,000; DAKO, Z0622).

Digits were fixed overnight in zinc-buffered formalin (Z-fix, Anatech, Battle Creek, MI). Bone was decalcified for 8 hours in a formic-acid based decalcifier (Decal I, Surgipath, Richmond, IL). Once decalcified, all samples were processed for paraffin embedding using a Leica TP 1020 Processor (Leica, Buffalo Grove, IL) and sectioned at 4 microns onto glass slides. Sections were stained with either Mayer’s Hematoxylin and Eosin Y, Mallory trichrome, or Picro-Sirius red stain (Sigma-Aldrich, St. Louis, MO) and mounted using permanent mounting medium (Fisher Scientific, Waltham, MA). Brightfield micrographs were captured using an Olympus DP72 camera mounted on an Olympus BX60 microscope with rotating stage (Olympus America Inc, Center Valley, PA). Polarized light microscopy was carried out with filters to provide circularly polarized illumination according to published methods [21 ]. Final figures were generated using Adobe Photoshop CS4.

Digits were fixed in zinc-buffered formalin (Z-fix, Anatech, Battle Creek, MI). Bone was decalcified for 48 hours in a formic-acid based decalcifier (Decal I, Surgipath, Richmond, IL). Once decalcified, all samples were processed for paraffin embedding and sectioned at 4 µm onto glass slides. Sections were stained with either Mayer’s Hematoxylin and Eosin Y, or Picro-Sirius red stain (Sigma-Aldrich, St. Louis, MO) and mounted using permanent mounting medium (Fisher Scientific, Waltham, MA). Brightfield micrographs were captured using an Olympus DP72 camera mounted on an Olympus BX60 microscope with rotating stage (Olympus America Inc, Center Valley, PA). Polarized light microscopy was carried out with filters to provide circularly polarized illumination.