Sample preparation
The Advanced Microscopy Facility provides sample preparation assistance both through service work, and by documentation made available on this page. Below is a list of the preparation techniques you can read about.
Disclaimer
Information in this Web site is intended to assist investigators in utilizing sample preparation methodology. Protocols have been reviewed for completeness but not tested for results. They should be considered a starting point. We cannot take responsibility for results or guarantee success. Success depends on many factors including sample, equipment, and technique. Those using these protocols are encouraged to contact the contributors directly if they have questions.
A Focused Ion Beam system is used to prepare TEM samples, section samples for SEM imaging, sensor and other apparatus fabrication, and many other tasks.
Online articles
- Focused ion beam on Wikipedia.
- University logo carved onto human hair with focused ion beam
- Focused Ion Beam at IBM's Almaden Research Center
- Applications of focused ion beam
- An article on a TEM specimen liftout procedure
Selected papers
- Takeo Tanaka, Tsuyoshi Irisawa, Makoto Fujiwara, and Norio Gennai, Micrometer-scale fabrication and assembly using focused ion beam, Thin solid films volume 509 pp 113 (2006)
- P.R. Munroe, The application of focused ion beam microscopy in the material sciences, Materials Characterization volume 60 pp 2 (2009)
- H.C. Floresca, J Jeon, J.G. Wang, and M.J. Kim, The Focused Ion beam Fold-Out: Sample Preparation Method for Transmission Electron Microscopy, Microscopy and Microanalysis volume 15 number 6 pp 558-563 (2009)
- M. Kammler, R. Hull, M.C. Reuter, and F.M. Foss, Lateral control of self-assembled island nucleation by focused-ion-beam micropatterning, Applied Physics Letters volume 82 number 7 pp 1093-1095 (2003)
- Peter J. Heaney, Edward P. Vicenzi, Lucille A. Giannuzzi, and Kenneth J.T. Livi,Focused ion beam milling: A method of site-specific sample extraction for microanalysis of Earth and planetary materials, American Mineralologist volume 86 number 9 pp 1094-1099 (2001)
- G.M. Shedd, H. Lezec, A.D. Dubner, and J. Melngailis, Focused ion beam induced deposition of gold, Applied Physics Letters volume 49 number 23 pp 1584-1586 (1986)
- J. A. Veerman, A. M. Otter, L. Kuipers, and N. F. van Hulst, High definition aperture probes for near-field optical microscopy fabricated by focused ion beam milling, Applied Physics Letters volume 72 number 24 pp 3115-3117 (1998)
- Gang Xiong, D.A. Allwood, M. D. Cooke, and R. P. Cowburn, Magnetic nanoelements for magnetoelectronics made by focused-ion-beam milling, Applied Physics Letters volume 79 number 21 pp 3461-3463 (2001)
- Thoru Ishitani, Hideki Tsuboi, Toshie Yaguchi, and Hidemi Koike, Transmission Electron Microscope Sample Preparation Using a Focused Ion Beam, Journal of Electron Microscopy volume 43 number 5 pp 322-326 (1994)
- Steve Reyntjens and Robert Puers, A review of focused ion beam applications in microsystem technology, Journal of Micromechanics and Microengineering volume 11 number 4 pp 287-300 (2001)
- Y.Z. Huang, S. Lozano-Perez, R.M. Langford, J.M. Titchmarsh, and M.L. Jenkins,Preparation of transmission electron microscopy cross-section specimens of crack tips using focused ion beam milling, Journal of Microscopy volume 207 number 2 pp 129-136 (2002)
- L.A. Giannuzzi and F.A. Stevie, A review of focused ion beam milling techniques for TEM specimen preparation, Micron volume 30 number 3 pp 197-204 (1999)
- M. W. Phaneuf, Applications of focused ion beam microscopy to materials science specimens, Micron volume 30 number 3 pp 277-288 (1999)
- Shinji Matsui and Yukinori Ochiai, Focused ion beam applications to solid state devices, Nanotechnology volume 7 number 3 pp 247-258 (1996)
- David P. Adams, Michael J. Vasile, Gilbert Benavides and Ann N. Campbell,Micromilling of metal alloys with focused ion beam–fabricated tools, Precision Engineering volume 25 number 2 pp 107-113 (2001)
- D. J. Larson, D. T. Foord, A. K. Petford-Long, H. Liew, M. G. Blamire, A. Cerezo and G. D. W. Smith, Field-ion specimen preparation using focused ion-beam milling, Ultramicroscopy volume 79 number 1-4 pp 287-293 (1999)
- P.D. Prewett and E.M. Kellogg, Liquid metal ion sources for FIB microfabrication systems - recent advances, Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms
- Herman Carlo Floresca, Jangbae Jeon, Jinguo G. Wang and Moon J. Kim, The Focused Ion Beam Fold-Out: Sample Preparation Method for Transmission Electron Microscopy, Microscopy and Microanalysis volume 15 number 6 pp 558-563 (2009)
- K.C. Wong, C.M. Haslauer, N. Anantharamaiah, B. Poiurdeyhima, A.D. Batchelor and D.P. Griffis, Focused Ion Beam Characterization of Bicomponent Polymer Fibers, Microscopy and Microanalysis volume 16 number 3 pp 282-290 (2010)
- Stefano Rubino, Sultan Akhtar, Petter Melin, Andrew Searle, Paul Spellward, Klaus Leifer, A site-specific focused-ion-beam lift-out method for cryo Transmission Electron Microscopy, Journal of Structural Biology Volume 180, Issue 3, pp 572–576 (2012)
- Jörn T. Wätjen, Jonathan J. Scragg, Marika Edoff, Stefano Rubino, Charlotte Platzer-Björkman, Cu out-diffusion in kesterites—A transmission electron microscopy specimen preparation artifact, Applied Physics Letters 102 (5), 051902 (2013)
- Stefano Rubino, Petter Melin, Paul Spellward, Klaus Leifer, Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam, Journal of Visualized Experiments (89):e51463 (2014)
Microwave processing systems are used to prepare many types of biological samples for electron microscopy, paraffin embedding, immunolabeling, in situ hybridization, histological stains, and epitope retrieval, to name a few.
Learn about the terminology used when discussing biological sample preparation.
Conventional methods which are used to prepare biological samples for SEM imaging and EDX analysis.
Every step of the process affects the quality of the final electron micrograph. There are eight major steps to prepare a biological SEM specimen:
- Primary fixation
- Washing
- Secondary fixation
- Dehydration / exchange of water in cell for 100% ethanol
- Exchange of ethanol with liquid CO2
- Critical point drying
- Mounting
- Sputter coating
The documents below show variations of this basic method, as well as supporting methods.
Conventional methods which are used to prepare biological samples for TEM imaging.
Every step of the process affects the quality of the final electron micrograph. There are nine major steps to preparing a biological TEM specimen:
- Primary fixation
- Washing
- Secondary fixation
- Enbloc staining
- Dehydration
- Infiltration with transitional solvents
- Infiltration with resin
- Embedding
- Curing
The documents below show variations of this basic method, as well as supporting methods such as sectioning, immunolabelling and staining.
- Grid preparation
- Resin preparation
- Protocol for animal tissue
- Protocol for plant tissue
- Protocol for cells on a substrate
- TEM sectioning
- TEM staining
- Immunolabelling
We also recommend the following external links: