Laser Capture Microdissection

Lu Charboneau1, Cloud P. Paweletz1, Lance A. Liotta1

1 National Institute of Health, National Cancer Institute, Bethesda, Maryland
Publication Name:  Current Protocols in Cell Biology
Unit Number:  Unit 2.5
DOI:  10.1002/0471143030.cb0205s10
Online Posting Date:  May, 2001
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Abstract

This unit describes laser capture microdissection (LCM) using the Pixcell II as a technique to provide the scientific community with the opportunity to perform molecular analyses on pure cell populations procured directly from tissues. After identifying specific cells of interest, the cells are captured by firing a near infrared laser through a thermaplastic polymer film that rests on top of the cells. The cells are then ready for molecular analyses.

     
 
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Table of Contents

  • Laser Capture Microdissection
  • Basic Protocol 1: Isolation of a Pure Cell Population from Tissue Sections
  • Support Protocol 1: Hematoxylin and Eosin Staining of Tissues for LCM
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Isolation of a Pure Cell Population from Tissue Sections

  Materials
  • Stained tissue samples, either frozen or formalin paraffin‐embedded, cut into 2‐ to 10‐µm sections, and mounted on plain, uncharged microscope slides
  • PixCell II Laser Capture Microdissection System (Arcturus Engineering)
  • CapSure transfer film (Arcturus Engineering)
  • Compressed gas duster
  • CapSure pads (Arcturus Engineering)
  • Cap removal tool (Arcturus Engineering)
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Figures

Videos

Literature Cited

Literature Cited
   Banks, R.E., Dunn, M.J., Forbes, M.A., Stanley, A., Pappin, D., Naven, T., Gough, M., Harnden, P., and Selby, P.J. 1999. The potential use of laser capture microdissection to selectively obtain distinct populations of cells for proteomic analysis—preliminary findings. Electrophoresis 20:689‐700.
   Emmert‐Buck, M.R., Bonner, R.F., Smith, P.D., Chuaqui, R.F., Zhuang, Z., Goldstein, S.R., Weiss, R.A., and Liotta, L.A. 1996. Laser capture microdissection. Science 274:998‐1001.
   Krizman, D.B., Chuaqui, R.F., Meltzer, P.S., Trent, J.M., Duray, P.H., Linehan, W.M., Liotta, L.A., and Emmert‐Buck, M.R. 1996. Construction of a representative cDNA library from prostatic intraepithelial neoplasia. Cancer Res. 56:5380‐5383.
   Simone, N.L., Bonner, R.F., Gillespie, J.W., Emmert‐Buck, M.R., and Liotta, L.A. 1998. Laser‐capture microdissection: Opening the microscopic frontier to molecular analysis. Trends Genet. 14:272‐276.
   Simone, N.L., Remaley, A.T., Charboneau, L., Petricoin, E.F. III, Glickman, J.W., Emmert‐Buck, M.R., Fleisher, T.A., and Liotta, L.A. 2000. Sensitive immunoassay of tissue cell proteins procured by laser capture microdissection. Am. J. Pathol. 156:445‐452.
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