High resolution three-dimensional histological reconstruction of glioma spheroids and application in research

Anke Bruning-Richardson, Nicholas Roberts, Doreen Crellin, Darren Treanor, Susan C Short

Research output: Contribution to journalMeeting Abstract

Abstract

BACKGROUND: Glioblastomas are highly malignant tumours characterized by an invasive phenotype, which limits response to standard treatment including surgery, radiation and chemotherapy. Targeting the migratory properties of glioma cells may offer an alternative means to improve disease management. We recently developed a 96 well-based in vitro collagen invasion assay to study the effect of anti-migratory drugs on glioma spheroids generated from patient derived and established cell lines. One limitation of this 3D technique is the quantitative assessment of drug activity as data is subjected to 2D analysis. Here we explored the feasibility of analysing spheroids and migratory cells using 3D image reconstruction. METHODS: Glioma spheroids (U87) were allowed to migrate into a collagen matrix over 72h and prepared for immunohistochemistry. Paraffin embedded spheroids were serially sectioned at 5 µM. A total of 50 slides per spheroid were stained using H&E for reconstruction of the whole spheroid and surrounding migratory rim. Slides were scanned with Aperio AT scanner at 20x magnification, and custom image registration software was used to generate and view 3D volumes. This software uses a novel method for slice-to-slice image registration using automatic registration algorithms custom designed for virtual slides and histopathology images RESULTS: By visualizing the fixed spheroids in 3D, we determined the number, morphology and distance traversed by migratory cells. We also ascertained differences between core and migrating cell morphology, number of cells and voxel sizes. CONCLUSIONS: We have demonstrated for the first time the feasibility of generating 3D histological reconstruction of glioma spheroids and migratory cells encased in collagen including whole slide scanning, image serving, registration and visualization in one procedure. Analysis of the glioma spheroids and the migratory cells allowed the quantitative analysis of research-relevant parameters. This technology can be used to analyse migratory behavior of glioma cells and drug activity in a 3D setting.
LanguageEnglish
Pages139-140
Number of pages2
JournalNeuro-Oncology
Volume16
Issue numberS5
DOIs
Publication statusPublished - Nov 2014
Externally publishedYes
Event19th Annual Scientific Meeting of the Society for Neuro-Oncology - Loews Hotel South Beach, Miami, United States
Duration: 13 Nov 201416 Nov 2014
Conference number: 19

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Glioma
Research
Collagen
Software
Pharmaceutical Preparations
Computer-Assisted Image Processing
Glioblastoma
Disease Management
Cell Size
Paraffin
Cell Count
Immunohistochemistry
Radiation
Technology
Phenotype
Drug Therapy
Cell Line
Neoplasms

Cite this

Bruning-Richardson, Anke ; Roberts, Nicholas ; Crellin, Doreen ; Treanor, Darren ; Short, Susan C. / High resolution three-dimensional histological reconstruction of glioma spheroids and application in research. In: Neuro-Oncology. 2014 ; Vol. 16, No. S5. pp. 139-140.
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title = "High resolution three-dimensional histological reconstruction of glioma spheroids and application in research",
abstract = "BACKGROUND: Glioblastomas are highly malignant tumours characterized by an invasive phenotype, which limits response to standard treatment including surgery, radiation and chemotherapy. Targeting the migratory properties of glioma cells may offer an alternative means to improve disease management. We recently developed a 96 well-based in vitro collagen invasion assay to study the effect of anti-migratory drugs on glioma spheroids generated from patient derived and established cell lines. One limitation of this 3D technique is the quantitative assessment of drug activity as data is subjected to 2D analysis. Here we explored the feasibility of analysing spheroids and migratory cells using 3D image reconstruction. METHODS: Glioma spheroids (U87) were allowed to migrate into a collagen matrix over 72h and prepared for immunohistochemistry. Paraffin embedded spheroids were serially sectioned at 5 µM. A total of 50 slides per spheroid were stained using H&E for reconstruction of the whole spheroid and surrounding migratory rim. Slides were scanned with Aperio AT scanner at 20x magnification, and custom image registration software was used to generate and view 3D volumes. This software uses a novel method for slice-to-slice image registration using automatic registration algorithms custom designed for virtual slides and histopathology images RESULTS: By visualizing the fixed spheroids in 3D, we determined the number, morphology and distance traversed by migratory cells. We also ascertained differences between core and migrating cell morphology, number of cells and voxel sizes. CONCLUSIONS: We have demonstrated for the first time the feasibility of generating 3D histological reconstruction of glioma spheroids and migratory cells encased in collagen including whole slide scanning, image serving, registration and visualization in one procedure. Analysis of the glioma spheroids and the migratory cells allowed the quantitative analysis of research-relevant parameters. This technology can be used to analyse migratory behavior of glioma cells and drug activity in a 3D setting.",
author = "Anke Bruning-Richardson and Nicholas Roberts and Doreen Crellin and Darren Treanor and Short, {Susan C}",
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doi = "10.1093/neuonc/nou264.9",
language = "English",
volume = "16",
pages = "139--140",
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High resolution three-dimensional histological reconstruction of glioma spheroids and application in research. / Bruning-Richardson, Anke; Roberts, Nicholas; Crellin, Doreen; Treanor, Darren; Short, Susan C.

In: Neuro-Oncology, Vol. 16, No. S5, 11.2014, p. 139-140.

Research output: Contribution to journalMeeting Abstract

TY - JOUR

T1 - High resolution three-dimensional histological reconstruction of glioma spheroids and application in research

AU - Bruning-Richardson, Anke

AU - Roberts, Nicholas

AU - Crellin, Doreen

AU - Treanor, Darren

AU - Short, Susan C

PY - 2014/11

Y1 - 2014/11

N2 - BACKGROUND: Glioblastomas are highly malignant tumours characterized by an invasive phenotype, which limits response to standard treatment including surgery, radiation and chemotherapy. Targeting the migratory properties of glioma cells may offer an alternative means to improve disease management. We recently developed a 96 well-based in vitro collagen invasion assay to study the effect of anti-migratory drugs on glioma spheroids generated from patient derived and established cell lines. One limitation of this 3D technique is the quantitative assessment of drug activity as data is subjected to 2D analysis. Here we explored the feasibility of analysing spheroids and migratory cells using 3D image reconstruction. METHODS: Glioma spheroids (U87) were allowed to migrate into a collagen matrix over 72h and prepared for immunohistochemistry. Paraffin embedded spheroids were serially sectioned at 5 µM. A total of 50 slides per spheroid were stained using H&E for reconstruction of the whole spheroid and surrounding migratory rim. Slides were scanned with Aperio AT scanner at 20x magnification, and custom image registration software was used to generate and view 3D volumes. This software uses a novel method for slice-to-slice image registration using automatic registration algorithms custom designed for virtual slides and histopathology images RESULTS: By visualizing the fixed spheroids in 3D, we determined the number, morphology and distance traversed by migratory cells. We also ascertained differences between core and migrating cell morphology, number of cells and voxel sizes. CONCLUSIONS: We have demonstrated for the first time the feasibility of generating 3D histological reconstruction of glioma spheroids and migratory cells encased in collagen including whole slide scanning, image serving, registration and visualization in one procedure. Analysis of the glioma spheroids and the migratory cells allowed the quantitative analysis of research-relevant parameters. This technology can be used to analyse migratory behavior of glioma cells and drug activity in a 3D setting.

AB - BACKGROUND: Glioblastomas are highly malignant tumours characterized by an invasive phenotype, which limits response to standard treatment including surgery, radiation and chemotherapy. Targeting the migratory properties of glioma cells may offer an alternative means to improve disease management. We recently developed a 96 well-based in vitro collagen invasion assay to study the effect of anti-migratory drugs on glioma spheroids generated from patient derived and established cell lines. One limitation of this 3D technique is the quantitative assessment of drug activity as data is subjected to 2D analysis. Here we explored the feasibility of analysing spheroids and migratory cells using 3D image reconstruction. METHODS: Glioma spheroids (U87) were allowed to migrate into a collagen matrix over 72h and prepared for immunohistochemistry. Paraffin embedded spheroids were serially sectioned at 5 µM. A total of 50 slides per spheroid were stained using H&E for reconstruction of the whole spheroid and surrounding migratory rim. Slides were scanned with Aperio AT scanner at 20x magnification, and custom image registration software was used to generate and view 3D volumes. This software uses a novel method for slice-to-slice image registration using automatic registration algorithms custom designed for virtual slides and histopathology images RESULTS: By visualizing the fixed spheroids in 3D, we determined the number, morphology and distance traversed by migratory cells. We also ascertained differences between core and migrating cell morphology, number of cells and voxel sizes. CONCLUSIONS: We have demonstrated for the first time the feasibility of generating 3D histological reconstruction of glioma spheroids and migratory cells encased in collagen including whole slide scanning, image serving, registration and visualization in one procedure. Analysis of the glioma spheroids and the migratory cells allowed the quantitative analysis of research-relevant parameters. This technology can be used to analyse migratory behavior of glioma cells and drug activity in a 3D setting.

U2 - 10.1093/neuonc/nou264.9

DO - 10.1093/neuonc/nou264.9

M3 - Meeting Abstract

VL - 16

SP - 139

EP - 140

JO - Neuro-Oncology

T2 - Neuro-Oncology

JF - Neuro-Oncology

SN - 1522-8517

IS - S5

ER -