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Integrated Biometal Science View Online Volume 1 Volume NANOCHEMISTRY New Edition of | Leading Textbook Number 2 | NANOCHEMISTRY 2009 Metallomics A Chemical Approach to Nanomaterials Integrated biometal science Geoffrey A Ozin, André C Arsenault and Ludovico Cademartiri www.rsc.org/metallomics Volume 1 | Number 2 | March 2009 | Pages 117–176 Metallomics This up-date of the globally successful 1st Edition highlights the latest breakthroughs using new case histories, problems and teaching principles. Extracts of reviews from the 1st Edition “A gem in the scientific literature...a beautifully written and richly illustrated book that is unlike any other.” Downloaded on 19 February 2011 Science, 21 July 2006, Vol 313 “An invaluable reference book for undergraduate and Published on 13 February 2009 http://pubs.rsc.org | doi:10.1039/B901310P graduate students. As a superb textbook for teaching of materials chemistry and nanotechnology.” Advanced Materials, 1/2006 “...wonderful book...insightful perspective on nanochemistry.” Physical Sciences Educational Reviews, October 2006, Vol. 7, Issue 2 Hardback | 876 pages | ISBN 9781847558954 | 2008 | £45.00 020963 Pages ISSN 1756-5901 www.rsc.org/books PAPER MINIREVIEW Registered Charity Number 207890 117–176 Doble et al. Ortega Elemental bio-imaging of calcium Synchrotron radiation for direct phosphate crystal deposits in knee analysis of metalloproteins on samples from arthritic patients electrophoresis gels 1756-5901(2009)1:2;1-5 View Online PAPER www.rsc.org/metallomics | Metallomics Elemental bio-imaging of calcium phosphate crystal deposits in knee samples from arthritic patients Christine Austin,a Dominic Hare,a Andrew L. Rozelle,b William H. Robinson,b Rudolf Grimmc and Philip Doble*a Received 20th January 2009, Accepted 20th January 2009 First published as an Advance Article on the web 13th February 2009 DOI: 10.1039/b901310p Laser ablation inductively coupled plasma mass spectrometry (LA ICP-MS) was employed to image deposits of calcium phosphate based crystals in knee cartilage and synovial fluid from arthritic patients. A reaction/collision cell containing hydrogen minimised plasma interferences on calcium and also improved the image quality without significant sensitivity reduction. Areas of high calcium and phosphorus intensities consistent with crystal deposits were observed for both the cartilage and synovial fluid samples. These areas were also characterised by high magnesium and strontium intensities. Distribution patterns of other elements such as copper and sulfur did not correlate with the crystal deposits. Filtered and non-filtered solutions of calcium phosphate crystals grown in synthetic synovial fluid were also imaged as further evidence of crystal deposits. The crystal deposits were detected in the unfiltered solution, and were absent from the filtered solutions. Introduction Other techniques for the determination of BCP crystals include scanning electron microscopy (SEM) and transmission The importance of calcium pyrophosphate dihydrate (CPPD) electron microscopy (TEM). These methods are expensive, and basic calcium phosphates (BCP) in rheumatology was first complex and not widely available for routine analysis in a 1 discovered in the early 1960s. Since then, the concurrence of clinical setting. Techniques recently applied to crystal detection BCP (hydroxyapatite, octacalcium phosphate, tricalcium and identification include atomic force microscopy (AFM), phosphate) and CPPD crystals, and degenerative joint disease Fourier-transform infrared spectroscopy (FTIR) and Raman Downloaded on 19 February 2011 has been well established. Though there is ample data spectroscopy. AFM and Raman show great potential for this available to support the role of BCP crystals in cartilage application. Additionally, FTIR and Raman spectrometers degeneration, it is still unclear whether other calcium-containing may be made compact, making the technology capable of crystals play a direct driving role in disease conception and 3 Published on 13 February 2009 http://pubs.rsc.org | doi:10.1039/B901310P point-of-care operation. A comprehensive review of the 2–4 progression, or are merely markers of joint damage. analytical tools available for the detection and identification The identification of specific crystal types in synovial fluid is of calcium phosphate crystals was recently published by currently the most popular method of diagnosing all common Yavorskyy et al.3 The review made it evident that a simple, 3 forms of crystal-associated arthritis. In general, crystal inexpensive technique which is capable of detecting and identi- arthritides cannot be distinguished from other causes of fying calcium phosphate-based crystals accurately and with a 5 arthritis based on history or physical examination. Despite high sensitivity would be invaluable in the diagnosis of crystal the multitude of techniques that have been applied to BCP and associated arthritides. CPPD crystal detection, clinical diagnosis almost exclusively Laser ablation inductively coupled plasma mass spectro- relies on light microscopy (LM). However, LM lacks the metry (LA ICP-MS) is increasingly applied to the study of resolution required to image small crystals (o1 mm), is non- metals in biological samples.8,9 LA ICP-MS offers direct specific (though CPPD crystals can generally be distinguished multielemental analysis of solids and liquids with trace and 3,5–7 under polarised LM) and accuracy is operator dependent. ultra trace detection capabilities, and a linear dynamic range Sensitivity can be improved by staining the sample with of seven or more orders of magnitude. Though its application calcium specific dyes, particularly for BCP crystals, which to imaging of soft tissues is emerging, it is considered a mature are often too small to be seen by LM. However these are technique for the analysis of geological and metallurgical often subject to false positives. samples.10 In LA ICP-MS systems, a focused laser beam is used to mobilise sample material as droplets or vapour from a Elemental Bio-imaging Facility, Department of Chemistry, Materials the sample surface.9 The material is then transported to the and Forensic Science, University of Technology Sydney, plasma (usually by argon carrier gas) where the material is Sydney NSW 2007, Australia. E-mail: [email protected]; Fax: +61 2 9514 1460; Tel: +61 2 9514 1792 ionised and carried through to the mass spectrometer, which b Division of Immunology and Rheumatology, Stanford University selectively detects ions at a given mass-to-charge ratio. School of Medicine, 300 Pasteur Drive, Stanford, LA ICP-MS may be utilised to construct maps that provide California 94305-5166, USA c Agilent Technologies, Inc., Integrated Biology Solutions Unit, elemental spatial information at trace and bulk levels 11 Santa Clara, California, USA of biological samples such as small tumours in rat brains, 142 | Metallomics, 2009, 1, 142–147 This journal is c The Royal Society of Chemistry 2009 View Online Table 1 Optimised experimental parameters for LA ICP-MS analysis Agilent 7500ce ICP-MS New wave UP213 laser ablation Rf power 1370 W Wavelength 213 nm Cooling gas flow rate 15 l minÀ1 Repetition frequency 20 Hz Carrier gas flow rate 1.25 l minÀ1 Laser energy density 0.2 J cmÀ2 (at 30%) Hydrogen flow rate 2.5 ml minÀ1 Cartilage samples Sample depth 5.0 mm Spot size 55 mm QP bias À16 V Scan rate 30 mmsÀ1 OctP bias À20 V Synovial fluid Scan mode Peak hopping Spot size 100 mm Dwell time 0.1 s, 0.2 s for P Scan rate 40 mmsÀ1 Synthetic synovial fluid Spot size 55 mm Scan rate 50 mmsÀ1 b-amyloid plaques in mouse models of Alzheimer’s disease,12 10 Â 10 mm) were imaged in approximately 20 h, whilst and the distribution of platinum in mouse kidneys treated with synovial fluid took approximately 2.5 h to image. cis-platin.13 The environmental sciences have also utilised the Imaging was performed using the software package ENVI potential of this technique to analyse the environmental v4.2 (Research Systems Inc., Boulder, CO, USA). The samples changes or pollutant accumulation evident from the changing were systematically scanned across the entire area of the metal concentrations in increment growth layers of many types sample. For cartilage samples, the distance between adjacent of organisms such as scales, shells, otoliths and tree xylem.14 scan lines was 29 mm, which ensured complete ablation of the Recently, Chaudhri et al.,15 determined the elemental tissue sample. For the synovial spots, the line scans were distribution of bladder and kidney stones by LA ICP-MS. placed 50 mm apart. The data files generated by ChemStation LA ICP-MS has also been employed to study element time (Agilent Technologies) were then collected and imported into profiles in teeth16 and bones,17 and to determine strontium ENVI. As no quantification data was collected, images only ratios in calcium phosphates.18 Quantitative analysis of heavy show relative signal intensities. metals in bone employing reference standards and spiking hydroxapatite standards has also been reported.17,19,20 However, LA ICP-MS has not been used to image crystal Sample preparation deposits in biological samples. Element concentrations in All experiments were performed in compliance with the relevant synovial fluids from osteoarthritic patients have been deter- Downloaded on 19 February 2011 laws and institutional guidelines of the University of Technology mined by ICP-MS.21 Sydney and Stanford University. The study was approved by the This study demonstrates the potential of LA ICP-MS for ethics committees of the University of Technology Sydney and crystal detection in biological samples. Cartilage sections were Published on 13 February 2009 http://pubs.rsc.org | doi:10.1039/B901310P Stanford University. Human samples were collected at Stanford obtained from knee or hip arthroplasty and were imaged by University after informed consent and under Institutional LA ICP-MS for the presence of crystal-associated elements. Review Board (IRB)-approved protocols. Cartilage sections were Additionally, eight synovial fluid samples, six from patients obtained after either knee or hip arthroplasty. Sections were cut diagnosed with osteoarthritis (OA) and two from patients to a thickness of 14 mm and then dried.
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