Scanning electron microscopy of bone: instrument, specimen, and issues

wbldb

Boyde, Alan¹; Jones, Sheila J.¹

Scanning electron microscopy of bone: instrument, specimen, and issues

Microsc Res Tech. February 1, 1996;33(2):92-120

©1996 Wiley-Liss, Inc

Affiliations

¹Department of Anatomy and Developmental Biology, University College London, London WC1E 6BT, United Kingdom
Abstract and keywords

There are many ways available now to maximise and analyse the information that can be obtained on the structure and constitution of bone using SEM. This paper considers a range of methods and the problems that arise relating to instrumentation and methodology as they apply to the use of SEM in the study of bone. In addition to the review content, some novel technical approaches to the SEM of bone are considered here for the first time; these include low kV imaging for the detection of new surface bone packets (and residual demineralized matrix after resorption), low kV BSE imaging of uncoated, embedded, and unembedded samples, environmental SEM for the study of wet tissue, low distortion, very low magnification imaging for the study of cancellous bone architecture, the use of multiple detectors for fast electrons in improving the imaging of porous samples, and high resolution, low voltage imaging for the study of collagen degradation during bone resorption.

Keywords: Scanning electron microscopy; Bone; Mineralisation; Resorption; Osteoporosis
Links

DOI: 10.1002/(SICI)1097-0029(19960201)33:2<92::AID-JEMT2>3.0.CO;2-0

PubMed: 8845522

WoS: A1996TN26600002
History

Received: 1994-02-16

Accepted: 1994-10-03

Cited Works (21)

Year	Entry
1971	Whitehouse WJ, Dyson ED, Jackson CK. The scanning electron microscope in studies of trabecular bone from a human vertebral body. J Anat. April 1971;108(3):481-496.
1993	Riggs CM, Lanyon LE, Boyde A. Functional associations between collagen fibre orientation and locomotor strain direction in cortical bone of the equine radius. Anat Embryol. March 1993;187(3):231-238.
1993	Grynpas M. Age and disease-related changes in the mineral of bone. Calcif Tiss Int. February 1993;53(suppl 1):S57-S64.
1993	Kneissel M. Vergleich von nicht-invasiven und invasiven Methoden zur Erfassung des alters- und geschlechtsabhängigen trabekulären Knochenverlustes an archäologischem Material [PhD thesis]. University of Vienna; 1993.
1969	Boyde A, Hobdell MH. Scanning electron microscopy of primary membrane bone. Z Zellforsch. 1969;99(1):98-108.
1988	Burr DB, Schaffler MB, Frederickson RG. Composition of the cement line and its possible mechanical role as a local interface in human compact bone. J Biomech. 1988;21(11):939-945.
1969	Boyde A, Hobdell MH. Scanning electron microscopy of lamellar bone. Z Zellforsch. 1969;93(2):213-231.
1990	Boyce TM, Bloebaum RD, Bachus KN, Skedros JG. Reproducible methods for calibrating the backscattered electron signal for quantitative assessment of mineral content in bone. Scanning Microsc. September 1990;4(3):591-600.
1986	Reid SA. A study of lamellar organisation in juvenile and adult human bone. Anat Embryol. 1986;174(3):329-338.
1987	Reid SA, Boyde A. Changes in the mineral density distribution in human bone with age: image analysis using backscattered electrons in the SEM. J Bone Miner Res. February 1987;2(1):13-22.
1994	Schaffler MB, Pitchford WC, Choi K, Riddle JM. Examination of compact bone microdamage using back-scattered electron microscopy. Bone. September–October 1994;15(5):483-488.
1993	Boyde A, Elliott JC, Jones SJ. Stereology and histogram analysis of backscattered electron images: age changes in bone. Bone. May–June 1993;14(3):205-210.
1984	Parfitt AM. Age-related structural changes in trabecular and cortical bone: cellular mechanisms and biomechanical consequences. Calcif Tiss Int. 1984;36(suppl 1):S123-S128.
1990	Mosekilde L. Consequences of the remodelling process for vertebral trabecular bone structure: a scanning electron microscopy study (uncoupling of unloaded structures). Bone Miner. July 1990;10(1):13-35.
1993	Mosekilde L. Vertebral structure and strength in vivo and in vitro. Calcif Tiss Int. February 1993;53(suppl 1):S121-S126.
1968	Baud CA. Submicroscopic structure and functional aspects of the osteocyte. Clin Orthop Relat Res. January–February 1968;56:227-236.
1969	Bélanger LF. Osteocytic osteolysis. Calcif Tiss Res. December 1969;4(1):1-12.
1993	Marotti G. A new theory of bone lamellation. Calcif Tiss Int. February 1993;53(suppl 1):S47-S56.
1986	Dempster DW, Shane E, Horbert W, Lindsay R. A simple method for correlative light and scanning electron microscopy of human iliac crest bone biopsies: qualitative observations in normal and osteoporotic subjects. J Bone Miner Res. February 1986;1(1):15-21.
1983	Boyde A, Jones SJ. Back-scattered electron imaging of skeletal tissues. Metab Bone Dis Rel Res. 1983;5(3):145-150.
1986	Boyde A, Maconnachie E, Reid SA, Delling G, Mundy GR. Scanning electron-microscopy in bone pathology: review of methods, potential and applications. Scan Electron Microsc. 1986;1986(4):1537-1554.

Cited By (25)

Year	Entry
2005	Skedros JG, Holmes JL, Vajda EG, Bloebaum AD. Cement lines of secondary osteons in human bone are not mineral‐deficient: new data in a historical perspective. Anat Rec. September 2005;286A(1):781-803.
2002	Benezra Rosen V, Hobbs LW, Spector M. The ultrastructure of anorganic bovine bone and selected synthetic hyroxyapatites used as bone graft substitute materials. Biomaterials. February 2002;23(3):921-928.
1997	Roschger P, Fratzl P, Klaushofer K, Rodan G. Mineralization of cancellous bone after alendronate and sodium fluoride treatment: a quantitative backscattered electron imaging study on minipig ribs. Bone. May 1997;20(5):393-397.
1997	Bloebaum RD, Skedros JG, Vajda EG, Bachus KN, Constantz BR. Determining mineral content variations in bone using backscattered electron imaging. Bone. May 1997;20(5):485-490.
1998	Boyde A, Compston JE, Reeve J, Bell KL, Noble BS, Jones SJ, Loveridge N. Effect of estrogen suppression on the mineralization density of iliac crest biopsies in young women as assessed by backscattered electron imaging. Bone. March 1998;22(3):241-250.
1998	Roschger P, Fratzl P, Eschberger J, Klaushofer K. Validation of quantitative backscattered electron imaging for the measurement of mineral density distribution in human bone biopsies. Bone. October 1998;23(4):319-326.
2004	Loveridge N, Power J, Reeve J, Boyde A. Bone mineralization density and femoral neck fragility. Bone. October 2004;35(4):929-941.
2009	Busse B, Hahn M, Soltau M, Zustin J, Püschel K, Duda GN, Amling M. Increased calcium content and inhomogeneity of mineralization render bone toughness in osteoporosis: mineralization, morphology and biomechanics of human single trabeculae. Bone. December 2009;45(6):1034-1043.
2010	Schneider P, Meier M, Wepf R, Müller R. Towards quantitative 3d imaging of the osteocyte lacuno-canalicular network. Bone. November 2010;47(5):848-858.
2014	Dong P, Haupert S, Hesse B, Langer M, Gouttenoire P-J, Bousson V, Peyrin F. 3D osteocyte lacunar morphometric properties and distributions in human femoral cortical bone using synchrotron radiation micro-CT images. Bone. March 2014;60:172-185.
2016	Mirzaali MJ, Schwiedrzik JJ, Thaiwichai S, Best JP, Michler J, Zysset PK, Wolfram U. Mechanical properties of cortical bone and their relationships with age, gender, composition and microindentation properties in the elderly. Bone. December 2016;93:196-211.
1999	Rinnerthaler S, Roschger P, Jakob HF, Nader A, Klaushofer K, Fratzl P. Scanning small angle X-ray scattering analysis of human bone sections. Calcif Tiss Int. May 1999;64(5):422-429.
2016	Hunt HB, Donnelly E. Bone quality assessment techniques: geometric, compositional, and mechanical characterization from macroscale to nanoscale. Clin Rev Bone Miner Metab. September 2016;14(3):133-149.
2003	Boyde A. The real response of bone to exercise. J Anat. August 2003;203(2):173-189.
2016	Georgiadis M, Müller R, Schneider P. Techniques to assess bone ultrastructure organization: orientation and arrangement of mineralized collagen fibrils. J R Soc Interface. June 1, 2016;13(119):20160088.
2005	Gupta HS, Schratter S, Tesch W, Roschger P, Berzlanovich A, Schoeberl T, Klaushofer K, Fratzl P. Two different correlations between nanoindentation modulus and mineral content in the bone–cartilage interface. J Struct Biol. February 2005;149(2):138-148.
2020	Madi K, Staines KA, Bay BK, Javaheri B, Geng H, Bodey AJ, Cartmell S, Pitsillides AA, Lee PD. In situ characterization of nanoscale strains in loaded whole joints via synchrotron X-ray tomography. Nat Biomed Eng. March 2020;4(3):343-354.
2017	Heveran CM. Bone Tissue Material Properties Are Altered in Chronic Kidney Disease to Lower Fracture Resistance Determining Bone Quality [PhD thesis]. Boulder, CO: University of Colorado; 2017.
2018	Hunt HB. Characterization of Material Properties, Microarchitecture, and Mechanics of Bone from Subjects With Type 2 Diabetes Mellitus [PhD thesis]. Ithaca, NY: Cornell University; December 2018.
2010	Feng L. Multi-Scale Characterization of Swine Femoral Cortical Bone and Long Bone Defect Repair by Regeneration [PhD thesis]. University of Illinois at Urbana-Champaign; 2010.
2016	Creighton E-R. Investigating Hyperglycemic Bone Formation With High Resolution Microscopy Techniques [Master's thesis]. Hamilton, ON: McMaster University; August 2016.
2011	Ahmed F. Multiscale Quantitative Imaging of Human Femoral Heads Using X-Ray Microtomography [PhD thesis]. Queen Mary University of London; 2011.
2012	Jimenez Palomar I. Mechanical Properties of Bone At the Sub-lamellar Level [PhD thesis]. London, England: Queen Mary University of London; September 2012.
2012	Tjhia CKH. Bone Quality of Patients With Atypical Fractures and Severely Suppressed Bone Turnover [PhD thesis]. Davis, University of California; 2012.
1998	Vajda EG. Age-Related Changes in the Microstructure and Mineralization of the Female Proximal Femur [PhD thesis]. University of Utah; March 1998.