3D image analysis reveals differences of CD30 positive cells and network formation in reactive and malignant human lymphoid tissue (classical Hodgkin Lymphoma).

Julia Liebers,Martin-Leo Hansmann,Kerstin Bianca Reisinger,Patrick Wurzel,Francesco Bertolini

doi:10.1371/journal.pone.0224156

Abstract

AimsThe examination of histological sections is still the gold standard in diagnostic pathology. Important histopathological diagnostic criteria are nuclear shapes and chromatin distribution as well as nucleus-cytoplasm relation and immunohistochemical properties of surface and intracellular proteins. The aim of this investigation was to evaluate the benefits and drawbacks of three-dimensional imaging of CD30+ cells in classical Hodgkin Lymphoma (cHL) in comparison to CD30+ lymphoid cells in reactive lymphoid tissues.Materials and resultsUsing immunoflourescence confocal microscopy and computer-based analysis, we compared CD30+ neoplastic cells in Nodular Sclerosis cHL (NScCHL), Mixed Cellularity cHL (MCcHL), with reactive CD30+ cells in Adenoids (AD) and Lymphadenitis (LAD). We confirmed that the percentage of CD30+ cell volume can be calculated. The amount in lymphadenitis was approx. 1.5%, in adenoids around 2%, in MCcHL up to 4,5% whereas the values for NScHL rose to more than 8% of the total cell cytoplasm. In addition, CD30+ tumour cells (HRS-cells) in cHL had larger volumes, and more protrusions compared to CD30+ reactive cells. Furthermore, the formation of large cell networks turned out to be a typical characteristic of NScHL.ConclusionIn contrast to 2D histology, 3D laser scanning offers a visualisation of complete cells, their network interaction and spatial distribution in the tissue. The possibility to differentiate cells in regards to volume, surface, shape, and cluster formation enables a new view on further diagnostic and biological questions. 3D includes an increased amount of information as a basis of bioinformatical calculations.

Highlights

In diagnostic pathology, the examination of immunostained, thin tissue sections using a lightmicroscope is considered to be the standard [1]
In contrast to 2D histology, 3D laser scanning offers a visualisation of complete cells, their network interaction and spatial distribution in the tissue
It seems worthwhile to evaluate whether a 3D laser scanning approach of thick histological sections can add additional valid data to conventional histology, and in how far it might be superior to methods routinely used [12]

Summary

Introduction

The examination of immunostained, thin tissue sections using a lightmicroscope is considered to be the standard [1] Digital visualisation of these sections called Whole Slide Images (WSI), enabled diagnosis on computer screens and was extended by bioinformatic methods [2]. Has become an integral part of diagnostics in clinical medicine, especially in radiology [6] Such radiological approaches provide a deeper insight in human organ structures and enable a more accurate diagnosis as well as precise planning of operations, improved tumor radiation and therapeutic tracer application [7,8,9,10]. It seems worthwhile to evaluate whether a 3D laser scanning approach of thick histological sections can add additional valid data to conventional histology, and in how far it might be superior to methods routinely used [12]

Methods

Results

Conclusion