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The sections were prepared using Masson's trichrome stain for the depiction of collagen [22].
Semi-thin (0.5 1.0 µm) sections were prepared using "Nova" ultra-tome LKB (Sweden), stained with Toluidine Blue and Methylene Blue-Basic Fuchsin.
Of this TMA 5 µm sections were prepared using silanized slides (Menzel Gläser, Braunschweig, Germany).
Thin sections were prepared using diamond knives (Diatome) on an MTX microtome (RM C Boeckeler Instruments, placed on 150-mesh coated copper grids (EMS) and stained with 3% uranyl acetate and lead citrate.
Negative control sections were prepared using normal mouse IgG instead of primary antibody.
Negative control sections were prepared using normal mouse IgG instead of the primary antibodies.
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From these samples, eleven cross-sections were prepared using different methods: 1. Sample KBr was pressed with KBr powder (Acros Organics, Belgium) into a conventional 13 mm diameter disk using Die Kit (PIKE Technologies, USA) and H-62 vacuum press (Trystom, Czech Republic) under 30 kN of pressure for 2 minutes.
Thin-sections were prepared using a Leica Ultracut UCT microtome (Leica Microsystems GmbH, Wetzlar, Germany).
After freezing of the OCT compound with tissues, 10 14µm cross-sections were prepared using a cryostat (Zeiss), and mounted on glass slides coated with a membrane film for dissection.
Ultrathin-sections were prepared using a Leica EM UC6 ultra-microtome.
Tissues were frozen in dry ice, and 16- μm serial cross-sections were prepared using a cryostat and collected on MAS-coated glass slides (Matsunami Glass, Osaka, Japan).
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