Table 8.

List of Tissues for Necropsy

Tissue	Processing at Necropsy
Tumor with brain	whole brain with tumor intact—tumor encompassed injection site; from target half of mice (random selection), whole brains (with tumor) flash-frozen in liquid nitrogen for qPCR; from target half of mice (random selection), whole brains (with tumor) placed into 10% NBF
Lymph nodes (LNs)	collected mandibular/cervical LNs as a single entity (as many as possible); left flash-frozen in liquid nitrogen for qPCR; right side placed into 10% NBF and stored for possible processing and histopathology
Spinal cord	cervical and thoracolumbar—(no bone in section) for qPCR; cervical and thoracolumbar—fix in situ, decalcified later for histopathology evaluation
Bone marrow (femur)	left femur—bone marrow flushed with phosphate-buffered saline and flash-frozen in liquid nitrogen for qPCR; right femur placed into 10% NBF and stored for processing and histopathology
Kidney, lungs, ovaries/testes	left side flash frozen in liquid nitrogen for qPCR; right side placed into 10% NBF and stored for processing and histopathology, with the exception of testes, which were preserved in modified Davidson’s fixative and subsequently transferred to 10% NBF according to testing facility SOP
Heart, liver, spleen	sample (~5 mm3) flash-frozen in liquid nitrogen for qPCR; remainder of tissue placed into 10% NBF and stored for processing and histopathology
Animal identification	collected but not processed for evaluation
Gross lesions (i.e., skin ulceration)	collected and processed as heart, liver, spleen, and retained at testing facility for possible qPCR and histopathology evaluation

NBF, neutral buffered formalin.