1. Spatial Distribution
of Copper and Iron in
Cardiac Tissue
Grant Allen

2. Seminar Outline
 Electron Probe Microanalysis
 Nuclear Microscopy
 Acknowledgements
 Secondary Ion Mass Spectrometry
 Conclusion
 Introduction

3. Introduction
 Investigate the spatial distribution of Cu and Fe in cardiac tissue
 Analytical techniques:
 Cardiac tissue that exhibits marked histological damage may
possess elevated levels of Cu and Fe
Electron probe x-ray microanalysis (EPMA)
Secondary ion mass spectrometry (SIMS)
Nuclear microscopy (NM)
 Correlate topographical features with chemical composition
 UHV techniques influence method of sample preparation

4. Electron Probe Microanalysis
Image courtesy of the Microscopy and Microanalysis Facility at the Department of Materials Engineering – Monash University
 Detection limit in the region of 100 ppm
 Primary ion beam: 5-20 kV electrons
 Lateral resolution of 1 µm
 Quantitative
 Cryochamber
 Specimen maintained at 80 K

5. Electron Probe Microanalysis
Energy /keV

6. Nuclear Microscopy
 Rutherford Backscattering Spectroscopy (RBS) - normalisation
 Scanning Transmission Ion Microscopy (STIM) - structural information
 Particle Induced X-ray Emission (PIXE) - elemental analysis
 Secondary electrons – complementary topographical information
 Incident beam: 1.0-3.0 MeV H+ or He+
 Lateral resolution of between 0.1 and 10 µm
 Detection limit: ppb to ppm
 Quantitative
 UHV chamber
 Specimen section freeze dried

7. Secondary Ion Mass Spectrometry
Image courtesy of the Bristol University CVD Diamond Group
 Detection limit: ppb to ppm
 Primary ion beam: 1-30 KeV 133Cs+
 Non-quantitative analysis of biological specimens
 Lateral resolution of 1 µm is possible
 UHV chamber
 Specimen section freeze dried

8. Secondary Ion Mass Spectrometry

9. Conclusion
 Nuclear microscopy: provided the specimen is prepared in an
appropriate manner, determination of the spatial distribution of
metals in biological tissue is possible
 Secondary ion mass spectrometry: non-quantitative
 Electron probe x-ray microanalysis: insufficient sensitivity

10.  Prof. Garth Cooper (Protemix)
Acknowledgements
 Dr Anthony Phillips (Protemix)
 Catherine Hobbis (School of Engineering - EPMA)
 Dr Marcus Gustafsson (Department of Chemistry - SIMS)
 Dr V. John Kennedy (Institute of Geological and Nuclear
Sciences - NM)
 Dr Ritchie Sims (Department of Geology - EPMA)