The above-mentioned remains were inside chamber O of the Ca’ Granda crypt. The access to the chamber was only possible from an opening in the ceiling and the manhole had remained sealed until the first opening. At that time, the archeologists accessed the chamber equipped with personal protective equipment to perform the first site inspection. After the preliminary inspection, the first excavation campaign begun. The remains were retrieved by the archaeologists, equipped with personal protective gear and gloves, under the supervision of the toxicologists. The crania, together with their preserved brain tissue, were placed in sterilized and sealed jars and left inside the Ca’ Granda crypt to maintain the same environmental conditions of the sepulcher chamber until the time of analysis. The post-cranial bones associated to the cranium C2 were also collected together with the cranium and placed in a separate sterilized box, in order to permit the anthropologists to study the entire skeleton. The samplings performed for each investigation (toxicological, radiocarbon, and histological investigations) were performed with sterilized scalpel and handsaw.
Well-preserved brain tissue samples were collected with a scalpel. The sample site was determined based on the consistency of the samples, choosing the best preserved area, as no specific zone of sampling is suggested for forensic toxicology specimen collection in such cases3 (link). For histological investigation, the section of sampling was selected based on the presence of well-preserved and well-visible encephalic convolutions, hypothesizing more preserved structures. For histological investigations three samples collected from the frontal, parietal and occipital brain area were selected.
The bone sample for the radiocarbon investigation was collected based on its weight and preservation, indeed the sample should weight about 5 g and be well-preserved with the cortical bone not affected by diagenesis or taphonomic processes. For toxicological investigations, the cranial sample was collected considering that the cranium was the only bone detected in the individuals under investigation apart from C2 that was associated with post-cranial bone samples. However, to perform a standardized sampling, we decided to collect only cranial samples, even if other bones were present (see post-cranial bone of C2). Moreover, the cranial sample can be considered a good bone matrix in forensic toxicology considering the excellent results obtained from previous studies that compared them with other bone sample sites5 (link),20 . The crania were cut with a sterilized handsaw on the occipital bone adjacent to the foramen magnum, and in cases where the foramen magnum was not preserved the parietal bone, in accordance with previous papers5 (link),20 .
Well-preserved brain tissue samples were collected with a scalpel. The sample site was determined based on the consistency of the samples, choosing the best preserved area, as no specific zone of sampling is suggested for forensic toxicology specimen collection in such cases3 (link). For histological investigation, the section of sampling was selected based on the presence of well-preserved and well-visible encephalic convolutions, hypothesizing more preserved structures. For histological investigations three samples collected from the frontal, parietal and occipital brain area were selected.
The bone sample for the radiocarbon investigation was collected based on its weight and preservation, indeed the sample should weight about 5 g and be well-preserved with the cortical bone not affected by diagenesis or taphonomic processes. For toxicological investigations, the cranial sample was collected considering that the cranium was the only bone detected in the individuals under investigation apart from C2 that was associated with post-cranial bone samples. However, to perform a standardized sampling, we decided to collect only cranial samples, even if other bones were present (see post-cranial bone of C2). Moreover, the cranial sample can be considered a good bone matrix in forensic toxicology considering the excellent results obtained from previous studies that compared them with other bone sample sites5 (link),20 . The crania were cut with a sterilized handsaw on the occipital bone adjacent to the foramen magnum, and in cases where the foramen magnum was not preserved the parietal bone, in accordance with previous papers5 (link),20 .
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