Skip to main content Accessibility help
×
Home

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

  • Asit Ranjan Mridha (a1), Indu Barwal (a2), Abhishek Gupta (a2), Abdul Majeed (a3), Adarsh W. Barwad (a1), Venkatesan Sampath Kumar (a3), Shivanand Gamanagatti (a4) and Subhash Chandra Yadav (a2)...

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.

Copyright

Corresponding author

*Author for correspondence: Subhash Chandra Yadav, E-mail: subhashmbu@gmail.com

References

Hide All
Anderson, TF (1951). Techniques for the preservation of three dimensional structure in preparing specimens for the electron microscope. Trans NY Acad Sci 13, 130134.
Aparisi, T (1978). Giant cell tumor of bone. Electron microscopic and histochemical investigations. Acta Orthop Scand Suppl 173, 138.
Arbeitsgemeinschaft, K, Becker, WT, Dohle, J, Bernd, L, Braun, A, Cserhati, M, Enderle, A, Hovy, L, Matejovsky, Z, Szendroi, M, Trieb, K & Tunn, PU (2008). Local recurrence of giant cell tumor of bone after intralesional treatment with and without adjuvant therapy. J Bone Joint Surg Am 90(5), 10601067.
Atkins, GJ, Kostakis, P, Vincent, C, Farrugia, AN, Houchins, JP, Findlay, DM, Evdokiou, A & Zannettino, AC (2006). RANK expression as a cell surface marker of human osteoclast precursors in peripheral blood, bone marrow, and giant cell tumors of bone. J Bone Miner Res 21(9), 13391349.
Balke, M, Schremper, L, Gebert, C, Ahrens, H, Streitbuerger, A, Koehler, G, Hardes, J & Gosheger, G (2008). Giant cell tumor of bone: Treatment and outcome of 214 cases. J Cancer Res Clin Oncol 134(9), 969978.
Bearer, EL & Orci, L (1986). A simple method for quick-freezing. J Electron Microsc Tech 3(2), 233241.
Boyde, A & Wood, C (1969). Preparation of animal tissues for surface-scanning electron microscopy. J Microsc 90(3), 221249.
Chakarun, CJ, Forrester, DM, Gottsegen, CJ, Patel, DB, White, EA & Matcuk, GR Jr. (2013). Giant cell tumor of bone: Review, mimics, and new developments in treatment. Radiographics 33(1), 197211.
Christine, D (1972). Preparation of alcohol-preserved larvae of Culicidae (Diptera) for scanning electron microscopy. Insect Syst Evol 3(3), 181188.
Dey, S (1993). A new rapid air-drying technique for scanning electron microscopy using tetramethylsilane: Application to mammalian tissue. Cytobios 73(292), 1723.
Frederik, PM & Busing, WM (1981). Ice crystal damage in frozen thin sections: Freezing effects and their restoration. J Microsc 121(Pt 2), 191199.
Gunning, WT & Crang, RE (1984). The usefulness of glutaraldehyde-carbohydrazidecopolymerization in biological specimen stabilization for scanning electron microscopy. J Electron Microsc Tech 1, 131140.
Hanaoka, H, Friedman, B & Mack, RP (1970). Ultrastructure and histogenesis of giant-cell tumor of bone. Cancer 25(6), 14081423.
Komiya, S (1982). Electron microscopy of bone tumors-osteosarcoma, chondrosarcoma, giant cell tumor of bone. Nihon Seikeigeka Gakkai Zasshi 56(7), 635657.
Liao, TS, Yurgelun, MB, Chang, SS, Zhang, HZ, Murakami, K, Blaine, TA, Parisien, MV, Kim, W, Winchester, RJ & Lee, FY (2005). Recruitment of osteoclast precursors by stromal cell derived factor-1 (SDF-1) in giant cell tumor of bone. J Orthop Res 23(1), 203209.
Morgan, T, Atkins, GJ, Trivett, MK, Johnson, SA, Kansara, M, Schlicht, SL, Slavin, JL, Simmons, P, Dickinson, I, Powell, G, Choong, PF, Holloway, AJ & Thomas, DM (2005). Molecular profiling of giant cell tumor of bone and the osteoclastic localization of ligand for receptor activator of nuclear factor kappaB. Am J Pathol 167(1), 117128.
Puri, A, Agarwal, MG, Shah, M, Jambhekar, NA, Anchan, C & Behle, S (2007). Giant cell tumor of bone in children and adolescents. J Pediatr Orthop 27(6), 635639.
Terracio, L & Schwabe, KG (1981). Freezing and drying of biological tissues for electron microscopy. J Histochem Cytochem 29(9), 10211028.
Zambo, I & Vesely, K (2014). WHO classification of tumours of soft tissue and bone 2013: The main changes compared to the 3rd edition. Cesk Patol 50(2), 6470.

Keywords

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

  • Asit Ranjan Mridha (a1), Indu Barwal (a2), Abhishek Gupta (a2), Abdul Majeed (a3), Adarsh W. Barwad (a1), Venkatesan Sampath Kumar (a3), Shivanand Gamanagatti (a4) and Subhash Chandra Yadav (a2)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed