Glue-Based Tissue Microarray Construction for Tumor and Disease Research

Summary

This protocol describes a low-cost production and efficient validation method for glue-based TMA construction, providing a convenient pathological diagnosis platform for tumor and disease research.

Abstract

Tissue microarray technology (TMA) is a high-throughput platform for the simultaneous detection and analysis of multiple tissue samples, facilitating efficient tumor and disease biomarker research. However, conventional TMA construction methods often face limitations such as operational complexity, time-consuming procedures, and variable accuracy. A glue-based TMA construction method was developed to overcome these challenges, offering improved tissue core fixation and enhanced structural stability. Systematic validation included histological evaluation (HE staining), immunohistochemical profiling of target proteins, and fluorescence in situ hybridization (FISH) analysis. Results demonstrated that the glue-based method maintained excellent slice integrity, improved signal-to-noise ratio, and ensured consistent batch-to-batch reproducibility. Although the approach is limited to manual operation, it presents a reliable and cost-effective option for moderate-throughput TMA production. This method is particularly suited to research environments that value flexibility and sample diversity over large-scale automation, expanding the utility of TMA in academic and diagnostic settings.

Introduction

Tissue Microarray (TMA) is a high-throughput technique for analyzing tissue samples^1,2,3. Multiple donor tissue cores are obtained, and donor paraffin blocks are transferred into recipient tissue blocks for simultaneous differential and comparative molecular analysis under theoretically the same performance conditions^2,4. The classic way to construct a tissue microarray (TMA) is to use a hole punch to extract tissue cores from a donor tissue sample and arrange them sequentially into a recipient paraffin block. This me....

Protocol

All donor blocks were obtained from archival pathological specimens collected between 2016 and 2018 at the Affiliated Huai'an No.1 People's Hospital of Nanjing Medical University. The samples were deidentified prior to use and processed in compliance with approved protocols (the ethics committee of the Affiliated Huai'an No.1 People's Hospital of Nanjing Medical University, KY-2024-250-01).

1. Assessment and tagging of donor tissue

1. Select tissue blocks with a thickness of ≥5 mm and an area of ≥15 mm × 15 mm. Ensure a 2 mm safety margin at the edges, and exclude regions exhibiting....

Results

In the present study, high-quality tissue microarrays were constructed using the glue method. To verify the effectiveness of the method, a series of experiments were performed, including H&E staining, immunohistochemical detection of specific proteins, and fluorescence in situ hybridization (FISH) analysis. A critical component of the construction process is the presence of tissue core dots at the expected positions and distances apart from one another, which is assessed by visual inspection. Visual inspecti.......

Discussion

As an innovative method for constructing tissue microarrays (TMA), the glue method has shown significant advantages due to its ease of construction procedures and cost-effectiveness. Compared to the traditional needle array method, which relies on sophisticated instruments^14,15, the glue method enables sample fixation by glueing, which can be done with only basic tools, greatly reducing the technical threshold. In contrast to the traditional embedding method, the.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Breast cancer HER2 Detection kit	Anbiping	2502001	Breast cancer HER2 Detection kit
CDHR4 antibody	Abcam	ab166914	CDHR4 antibody
CDK1 antibody	Abcam	ab265590	CDK1 antibody
CRTAC1 antibody	Abcam	ab254691	CRTAC1 antibody
DNASE1L3 antibody	Abcam	ab203669	DNASE1L3 antibody
Embedding machine	P.S.J MEDICAL	BM450A	Embedding machine
Fully automatic tissue dehydrator	Leica Biosystems	ASP3005	Fully automatic tissue dehydrator
Glass microscope slides	Citotest	250124A1	Glass microscope slides
Glue	TIZO	200	Glue
GPR146 antibody	Abcam	ab117104	GPR146 antibody
IGSF10 antibody	Abcam	ab197671	IGSF10 antibody
ITIH1 antibody	Abcam	ab233032	ITIH1 antibody
Low Profile Microtome Blades	Thermo Fisher	3052835	Low Profile Microtome Blades
Marker pen	Deli	SK109	Marker pen
Microtome	Leica Biosystems	HistoCore BIOCUT	Microtome
Paraffin wax	Solarbio	YA0012	Paraffin wax
SMAD9 antibody	Abcam	ab262940	SMAD9 antibody
TARBP1 antibody	Abcam	ab115896	TARBP1 antibody
ZCCHC24 antibody	Abcam	ab88756	ZCCHC24 antibody