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Processing Techniques for Scanning Electron Microscopy Imaging of Giant Cells from Giant Cell Tumors of Bone

Published online by Cambridge University Press:  30 August 2019

Asit Ranjan Mridha ,
Indu Barwal ,
Abhishek Gupta Open the ORCID record for Abhishek Gupta [Opens in a new window] ,
Abdul Majeed ,
Adarsh W. Barwad ,
Venkatesan Sampath Kumar ,
Shivanand Gamanagatti  and
Subhash Chandra Yadav
Asit Ranjan Mridha
Affiliation:
Department of Pathology, All India Institute of Medical Sciences, New Delhi 110029, India
Indu Barwal
Affiliation:
Department of Anatomy, All India Institute of Medical Sciences, New Delhi 110029, India
Abhishek Gupta
Affiliation:
Department of Anatomy, All India Institute of Medical Sciences, New Delhi 110029, India
Abdul Majeed
Affiliation:
Department of Orthopaedics, All India Institute of Medical Sciences, New Delhi 110029, India
Adarsh W. Barwad
Affiliation:
Department of Pathology, All India Institute of Medical Sciences, New Delhi 110029, India
Venkatesan Sampath Kumar
Affiliation:
Department of Orthopaedics, All India Institute of Medical Sciences, New Delhi 110029, India
Shivanand Gamanagatti
Affiliation:
Department of Radiodiagnosis, All India Institute of Medical Sciences, New Delhi 110029, India
Subhash Chandra Yadav*
Affiliation:
Department of Anatomy, All India Institute of Medical Sciences, New Delhi 110029, India
*
*Author for correspondence: Subhash Chandra Yadav, E-mail: subhashmbu@gmail.com
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Abstract

Giant cell tumor (GCT) of bone is a common benign lesion that causes significant morbidity due to the failure of modern medical and surgical treatment. Surface ultra-structures of giant cells (GCs) may help in distinguishing aggressive tumors from indolent GC lesions. This study aimed to standardize scanning electron microscopic (SEM) imaging of GC from GCT of bone. Fresh GCT collected in Dulbecco's Modified Eagle Medium was washed to remove blood, homogenized, or treated with collagenase to isolate the GCs. Mechanically homogenized and collagenase-digested GCs were imaged on SEM after commonly used drying methodologies such as air-drying, tetramethylsilane (TMS)-drying, freeze-drying, and critical point-drying (CPD) for the optimization of sample processing. The collagenase-treated samples yielded a greater number of isolated GC and showed better surface morphology in comparison to mechanical homogenization. Air-drying was associated with marked cell shrinkage, and freeze-dried samples showed severe cell damage. TMS methodology partially preserved the cell contour and surface structures, although the cell shape was distorted. GC images with optimum surface morphology including membrane folding and microvesicular structures on the surface were observed only in collagenase-treated and critical point-dried samples. Collagenase digestion and critical point/TMS-drying should be performed for optimal SEM imaging of individual GCs.

Keywords

boneGCTgiant cell tumorscanning electron microscopySEM of giant cell
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 25 , Issue 6 , December 2019 , pp. 1376 - 1382
Copyright
Copyright © Microscopy Society of America 2019 

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