Resolving Questions of Biological Interface Chemistry with TOF-SIMS and FIB-TOF Tomography
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1 Resolving Questions of Biological Interface Chemistry with TOF-SIMS and FIB-TOF Tomography Gregory L. Fisher, John S. Hammond & Scott R. Bryan, Physical Electronics With acknowledgements to: Prof. Reinhard Jetter, Dr. Phil Wong, Christopher Buschhaus at UBC Prof. Hans Lust, Dr. Priit Möller, Rait Kanarbik at Univ. of Tartu
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3 Why the Interest in Epicuticular Waxes? The cuticle is of ecological importance because it forms the interface between plant organs and the environment. The function(s) of plant cuticles can best be understood on the basis of the characteristic cuticular wax composition and the biosynthetic origin of the cuticular wax components.
4 Verification of Cuticular Wax Components e.g. Chemical Imaging of the Stem The vascular and cuticular regions are observed along with the substrate. Data removed due to pending publication.
5 Specialized Arabidopsis Cells spores stomates trichome SEM Images dev.biologists.org botanicalgarden.ubc.ca planttrichome.org TOF-SIMS Images Data removed due to pending publication.
6 Arabidopsis th. Abaxial Leaf & Stomates Data removed due to pending publication.
7 Chemical Characterization of Stomates Data removed due to pending publication.
8 Chemical Characterization of Trichomes Data removed due to pending publication.
9 Summary Specialized cells within a single organ appear to have distinct epicuticular wax compositions. Chemical imaging of intact organs provides verification that specific spectral features are, in fact, components of epicuticular wax. The observation and identification of specific wax components on specialized cuticular cells by TOF-SIMS is relatively straight forward. Such an analysis is difficult or impossible with more traditional extraction & chromatography or microscopy methods. These observations and chemical characterization are easily achieved owing to the unique attributes of the TRIFT analyzer. These results have been confirmed by duplicate analyses.
10 3D Imaging by FIB-TOF Tomography tomography sample preparation FIB sectioning TOF-SIMS ion & electron imaging image processing Sample stage is not moved between sectioning and ion/electron imaging. FIB Imaging Plane z y x LMIG
11 Why Use FIB-TOF Tomography? Need for isotopic, chemical and molecular analysis which is not possible with FIB-SEM/EDS or FIB-SIMS. Other advantages of TOF-SIMS surface and trace level sensitivity, analysis of insulators and dielectrics, fast data acquisition, full mass spectrum at every image pixel, & retrospective analysis. 3D imaging without the artifacts of sputter depth profiling differential sputtering loss of true 3D distributions accumulated beam damage loss of chemical / molecular info particles, pores & voids loss of true 3D distributions
12 3D FIB-TOF Tomography of CuW Composite LMIG-induced secondary electron (SE) images. The roughness observed ~ 10µm at the bottom of the FIB crater is the result of differential sputtering. Depth profile analysis would produce 3D images without the true distributions of W and Cu. 20 µm x 20 µm FOV. However, the elemental/chemical distribution is perfectly preserved in the FIB-milled section. Twenty (20) FIB-TOF tomography cycles at 0.5 µm each.
13 Profiles & 3D Images from FIB-TOF Data Imaged volume is 20 µm x 20 µm x 10 µm Counts Per Second 10 5 Total Ion WO Cu Depth (nm)
14 Why Study SOFCs? Understand growth conditions, materials and performance: Unintentional contaminants Layer and interfacial homogeneity; interlayer diffusion Optimized growth conditions Evaluate materials versus conversion efficiency Fuel cell geometry versus start-up time and operational efficiency Understand electro-chemical degradation over lifetime: Cathodic and electrolytic poisoning Change in electrolytic O 2- transport resistance versus temperature Goals to reduce the operational temperature (currently > 600 o C) and the start-up time (currently hours)
15 Initial FIB-Milled Crater in a SOFC LMIG-induced secondary electron (SE) images. (A) (B) (C) cathode electrolyte electrolyte anode PrSrCoOx Gd-doped CeOx Sc-stabilized ZrOx SE Image (after milling). SE Image (after polish). SE Image (after polish).
16 Measuring the FIB-Milled Z-Scale LMIG-induced secondary electron (SE) images. Z-direction 300 µm x 300 µm FOV.
17 Mass Spectral Characterization Data removed due to pending publication. With the TRIFT mass analyzer, the full mass spectrum is collected at every image pixel.
18 Identification of Matrix Components Isotopic abundances are used to confirm peak assignments. Data removed due to pending publication. Example showing identification of Sr, Y and Zr.
19 3D Tomography Data of a SOFC Imaged volume is 50 µm x 50 µm x 10 µm. 50 µm in X Data removed due to pending publication. 50 µm in Y 50 µm in Y 10 µm in Z 50 µm in X 10 µm in Z
20 3D FIB-TOF Imaging of a SOFC Imaged volume is 50 µm x 50 µm x 10 µm. Data removed due to pending publication. Sr +, Ce +, Zr + and K + 3D iso-surface overlays.
21 3D FIB-TOF Imaging of a SOFC Imaged volume is 50 µm x 50 µm x 10 µm.
22 Developments in Organic FIB-TOF Analysis of 10% Cocaine-Doped PLGA Spheres After FIB milling, can molecular ion signals be recovered by cluster ion sputter cleaning?
23 FIB Cut of Cocaine (10 wt.%)/plga Sphere Side view of FIB-milled sphere. Following high kv FIB milling. FIB cut direction
24 Depth Profile of Cocaine/PLGA Particle 20 kev C 60+ sputter removal of FIB straggle Cocaine [M+H] + image at surface. Counts Per Second Si (28m/z) Ca (40m/z) C 3 H 4 O (56m/z) C 2 H 3 (27m/z) Cocaine [M+H] + image below straggle region M+H (304m/z) Depth (nm)
25 Summary & Outlook TRIFT analyzer allows ion and electron imaging of a FIB-milled specimen without moving the sample from the milling position. This is advantageous for 3D imaging by FIB-TOF tomography. The TRIFT analyzer produces high sensitivity, useful mass resolution and high mass scale linearity for exploratory analysis. FIB milling of bulk insulators may be achieved, with or without a conductive capping layer. The need for post-mill FIB polishing is evident. Cluster ion sputtering may also be used to remove the FIB straggle. Work on 3D FIB-TOF imaging of cells and tissues is underway. The sample preparation, e.g. soft-matrix embedding, is found to be important.
26 PHI TRIFT V nanotof Thank You!
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