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

Asit Ranjan Mridha, Indu Barwal, Abhishek Gupta, Abdul Majeed, Adarsh W. Barwad, Venkatesan Sampath Kumar, Shivanand Gamanagatti, Subhash Chandra Yadav

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)1376-1382
Number of pages7
JournalMicroscopy and Microanalysis
Volume25
Issue number6
DOIs
StatePublished - 1 Dec 2019
Externally publishedYes

Keywords

  • GCT
  • SEM of giant cell
  • bone
  • giant cell tumor
  • scanning electron microscopy

ASJC Scopus subject areas

  • Instrumentation

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