A Standardized Protocol for the Safe Retrieval of Infectious Postmortem Human Brain for Studying Whole-Brain Pathology

James, Ranjit Immanuel, Verma, Richa, Johnson, Latif Rajesh, Manesh, Abi, Jayakumar, Jaikishan, Sen, Mousumi, Joseph, Jayaraj, Kumarasami, Ramdayalan, Mitra, Partha P, Sivaprakasam, Mohanasankar, Varghese, George M (July 2023) A Standardized Protocol for the Safe Retrieval of Infectious Postmortem Human Brain for Studying Whole-Brain Pathology. American Journal of Forensic Medicine and Pathology. ISSN 0195-7910

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Abstract

We describe a safe and standardized perfusion protocol for studying brain pathology in high-risk autopsies using a custom-designed low-cost infection containment chamber and high-resolution histology. The output quality was studied using the histological data from the whole cerebellum and brain stem processed using a high-resolution cryohistology pipeline at 0.5 μm per pixel, in-plane resolution with serial sections at 20-μm thickness. To understand the pathophysiology of highly infectious diseases, it is necessary to have a safe and cost-effective method of performing high-risk autopsies and a standardized perfusion protocol for preparing high-quality tissues. Using the low-cost infection containment chamber, we detail the cranial autopsy protocol and ex situ perfusion-fixation of 4 highly infectious adult human brains. The digitized high-resolution histology images of the Nissl-stained series reveal that most of the sections were free of processing artifacts, such as fixation damage, freezing artifacts, and osmotic shock, at the macrocellular and microcellular level. The quality of our protocol was also tested with the highly sensitive immunohistochemistry staining for specific protein markers. Our protocol provides a safe and effective method in high-risk autopsies that allows for the evaluation of pathogen-host interaction, the underlying pathophysiology, and the extent of the infection across the whole brain at microscopic resolutions.

Item Type:	Paper
Subjects:	organism description > animal organs, tissues, organelles, cell types and functions > organs types and functions > brain organism description > animal > mammal > primates > hominids organism description > animal > mammal > primates > hominids > human organism description > animal > mammal organs, tissues, organelles, cell types and functions > organs types and functions organs, tissues, organelles, cell types and functions organism description > animal > mammal > primates
CSHL Authors:	Mitra, Partha P.
Communities:	CSHL labs > Mitra lab
SWORD Depositor:	CSHL Elements
Depositing User:	CSHL Elements
Date:	25 July 2023
Date Deposited:	11 Oct 2023 15:02
Last Modified:	10 Jan 2024 19:37
PMCID:	PMC10662599
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/41177

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