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All animals were anaesthetized with sodium pentobarbital and perfused transcardially with freshly depolymerised 4% paraformaldehyde in 0.1 M phosphate buffer; the head was removed and further fixed for 24 h in the same fixative.
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The head was fixed with 10% neutral buffer formalin for 48 hours.
The heads were stored in phosphate buffered saline (PBS) for up to 5 days before preparation and testing.
For immunohistochemistry, marmosets that were terminally anesthetized (see above), were perfused transcardially with 0.9% saline, followed by 200 ml of fixative containing 4% paraformaldehyde in 0.1 M sodium-phosphate buffer (pH 7.2) for 15 min. The heads were postfixed in the same fixative, and brains were carefully removed from the skulls on the following day.
The heads were perfused with 3% glutaraldehyde in phosphate buffer overnight.
The heads were fixed for three hours at room temperature in 4% PBS (phosphate buffer saline) buffered (pH: 7.2) formaldehyde solution containing 0.25% Triton X-100 (Tx).
The heads were decalcified in 0.1 M EDTA in 0.1 M phosphate buffer.
After washing two times with DEPC-treated PBS buffer, half of the femoral head was immediately frozen in liquid nitrogen for future PCR and western blot analysis.
Similarly, contact pressures between the buffer and the metallic femoral head are shown in the prosthetic Tribofit model.
Upon receiving a buffer, the buffer is dissected and the iPod response is determined.
For bacterial culture from untreated heads, tonsillae were removed aseptically but singed heads were washed in Buffered Pepton Water and a sample of water taken for culture.
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