Spinal cord from body donors is suitable for multicolor immunofluorescence

Abstract

Immunohistochemistry is a powerful tool for studying neuronal tissue from humans at the molecular level. Obtaining fresh neuronal tissue from human organ donors is difficult and sometimes impossible. In anatomical body donations, neuronal tissue is dedicated to research purposes and because of its easier availability, it may be an alternative source for research. In this study, we harvested spinal cord from a single organ donor 2 h (h) postmortem and spinal cord from body donors 24, 48, and 72 h postmortem and tested how long after death, valid multi-color immunofluorescence or horseradish peroxidase (HRP) immunohistochemistry is possible. We used general and specific neuronal markers and glial markers for immunolabeling experiments. Here we showed that it is possible to visualize molecularly different neuronal elements with high precision in the body donor spinal cord 24 h postmortem and the quality of the image data was comparable to those from the fresh organ donor spinal cord. High-contrast multicolor images of the 24-h spinal cords allowed accurate automated quantification of different neuronal elements in the same sample. Although there was antibody-specific signal reduction over postmortem intervals, the signal quality for most antibodies was acceptable at 48 h but no longer at 72 h postmortem. In conclusion, our study has defined a postmortem time window of more than 24 h during which valid immunohistochemical information can be obtained from the body donor spinal cord. Due to the easier availability, neuronal tissue from body donors is an alternative source for basic and clinical research.

Distal Nerve Transfers in High Peroneal Nerve Lesions: An Anatomical Feasibility Study

“…It was not possible to limit counts to α-motoneurons, because ChAT signals were too weak in some donor nerves due to the postmortem interval (interval between the time of death of individuals and tissue fixation). This is in accordance with a recent study where we have demonstrated that the ChAT signal is stable for 24 h after death and then decreases [ 19 ]. The TA branch showed 1597.14 ± 325.94 myelinated fibers ( Table 1 ).…”

In focus in HCB

“…The question therefore remains: What is the allowable interval between death and tissue fixation to provide adequate samples for research purposes, especially those requiring immunohistochemistry? Reissig et al ( 2022 ) have now undertaken such a rigorous immunohistochemical study using human body donor spinal cord samples received 24 h, 48 h, and 72 h postmortem. The tissue immunostaining results from these samples were compared to those from a “control” sample organ donor received 2 h postmortem.…”