A novel frozen brain tissue array technique: immunohistochemical detection of neuronal paraneoplastic autoantibodies

Abstract

We introduce a modification of the tissue microarray technique in which several frozen brain tissue specimens are collected to a single frozen brain array block. In the present application, we use it for the detection of neuronal paraneoplastic anti-Hu autoantibodies. Representative samples from 15 different brain regions were collected according to a standard neuropathological autopsy protocol. Cryostat sections from each block were cut and conventionally stained. From representative areas, cylinder tissue samples from each specimen were punched and then arrayed into a recipient array block. Using the cryostat sections of this brain array, autoantibodies from seven anti-Hu-positive patient sera (confirmed by immunoblotting) were screened by immunohistochemistry. Neuronal architecture was well preserved and immunohistochemical staining was comparable to that of conventional cryostat sections. Because of the variable staining pattern in different brain areas, two anti-Hu-positive sera could be detected immunohistochemically by the one brain array. With the present array technique, it is possible to characterize the variable staining patterns of neuronal paraneoplastic autoantibodies in different locations of the human brain. The frozen brain array also allows the detection of RNA and DNA targets involved in neurological diseases.

Systematic immunohistochemical profiling of 378 brain tumors with 37 antibodies using tissue microarray technology

“…However, some reports applied TMAs to deoxyribonucleic acid (DNA) fluorescence in situ hybridization with paraffin blocks (TMAFISH) [2,5,9]. Furthermore, recent studies showed that TMAs could also be used for frozen tissue [4,13] and cell line [6] analyses.…”

Tissue microarrays enabling high-throughput molecular pathology

“…Construction of frozen TMAs is challenging, largely because handling of the tissue and slides requires great care and storage of frozen TMA is cumbersome [18]. Overall the use of frozen TMAs is limited, typically for antibodies that do not perform in immunohistochemistry in paraffin-embedded tissue, or for in situ applications.…”

Tissue microarrays as a platform for proteomic investigation

“…Unfortunately large, well preserved, and well annotated collections of tissue are challenging to obtain as a source material. In addition, construction of frozen TMAs is challenging work and handling of the tissue and slides requires haste and great care (Kylaniemi et al 2004). Furthermore, storage of frozen TMAs is cumbersome.…”