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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

  •  on Wikipedia.
  • University logo carved onto human hair with 
  • Focused Ion Beam at 
  •  of focused ion beam
  • An  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, 
  • P.R. Munroe, The application of focused ion beam microscopy in the material sciences, 
  • 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, 
  • M. Kammler, R. Hull, M.C. Reuter, and F.M. Foss, Lateral control of self-assembled island nucleation by focused-ion-beam micropatterning, 
  • 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, 
  • G.M. Shedd, H. Lezec, A.D. Dubner, and J. Melngailis, Focused ion beam induced deposition of gold, 
  • 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, 
  • Gang Xiong, D.A. Allwood, M. D. Cooke, and R. P. Cowburn, Magnetic nanoelements for magnetoelectronics made by focused-ion-beam milling, 
  • Thoru Ishitani, Hideki Tsuboi, Toshie Yaguchi, and Hidemi Koike, Transmission Electron Microscope Sample Preparation Using a Focused Ion Beam, 
  • Steve Reyntjens and Robert Puers, A review of focused ion beam applications in microsystem technology, 
  • 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, 
  • L.A. Giannuzzi and F.A. Stevie, A review of focused ion beam milling techniques for TEM specimen preparation, 
  • M. W. Phaneuf, Applications of focused ion beam microscopy to materials science specimens, 
  • Shinji Matsui and Yukinori Ochiai, Focused ion beam applications to solid state devices, 
  • David P. Adams, Michael J. Vasile, Gilbert Benavides and Ann N. Campbell,Micromilling of metal alloys with focused ion beam–fabricated tools, 
  • 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, 
  • P.D. Prewett and E.M. Kellogg, Liquid metal ion sources for FIB microfabrication systems - recent advances, 
  • 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, 
  • 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, 
  • 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, 
  • 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, 
  • Stefano Rubino, Petter Melin, Paul Spellward, Klaus Leifer, Cryo-electron Microscopy Specimen Preparation By Means Of a Focused Ion Beam,

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:

  1. Primary fixation
  2. Washing
  3. Secondary fixation
  4. Dehydration / exchange of water in cell for 100% ethanol
  5. Exchange of ethanol with liquid CO2
  6. Critical point drying
  7. Mounting
  8. 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:

  1. Primary fixation
  2. Washing
  3. Secondary fixation
  4. Enbloc staining
  5. Dehydration
  6. Infiltration with transitional solvents
  7. Infiltration with resin
  8. Embedding
  9. Curing

The documents below show variations of this basic method, as well as supporting methods such as sectioning, immunolabelling and staining.

We also recommend the following external links:

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  • at .