Laser capture microdissection

Science

Volumn 274, Issue 5289, 1996, Pages 998-1001

  Emmert Buck, M R ^a   Bonner, R F ^b   Smith, P D ^b   Chuaqui, R F ^a   Zhuang, Z ^a   Goldstein, S R ^b   Weiss, R A ^a   Liotta, L A ^a  

^a NATIONAL CANCER INSTITUTE   (United States)

^b NATIONAL INSTITUTES OF HEALTH   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CARBON DIOXIDE LASERS; CELLS; DNA; ENZYMES; LASER PULSES; MICROSCOPIC EXAMINATION; POLYMERS; RNA; THERMOPLASTICS; THIN FILMS; TISSUE; VISUALIZATION;

ENZYME RECOVERY; ETHYLENE VINYL ACETATE POLYMER; HISTOPATHOLOGY; LASER CAPTURE MICRODISSECTION; THERMOPLASTIC FILM; TISSUE ANALYSIS;

LASER APPLICATIONS;

DNA; ENZYME; ETHYLENE VINYL ACETATE COPOLYMER; RNA;

ARTICLE; CARBON DIOXIDE LASER; CELL ADHESION; CELL POPULATION; DISSECTION; ENZYME ASSAY; GENETIC ANALYSIS; HUMAN; HUMAN TISSUE; POLYMERASE CHAIN REACTION; PRIORITY JOURNAL;

CELL SEPARATION; DISSECTION; DNA; FROZEN SECTIONS; HISTOLOGICAL TECHNIQUES; HUMANS; LASERS; PARAFFIN EMBEDDING; POLYMERASE CHAIN REACTION; POLYVINYLS; PRECANCEROUS CONDITIONS; RNA; TISSUE FIXATION;

EID: 0030575911     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.274.5289.998     Document Type: Article

References (25)

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13. Standard 6-μm sections from formalin- or alcohol-fixed, paraffin-embedded archival tissue samples were prepared on noncoated glass slides. Sections were deparaffinized, stained with hematoxylin and eosin, treated with 3% glycerol in water for 1 min, and air-dried for 5 min before LCM. Fresh tissue was snap-frozen immediately after surgery at -70°C. Cryostat sections (6 μm) were prepared on standard glass histology slides. Tissue sections were fixed in formalin or alcohol and stained with hematoxylin and eosin (Lemer Laboratories, Pittsburgh, PA). Sections were dehydrated in graded alcohols and air-dried for 5 min before LCM transfer.
14. -1 at the laser wavelength (10.6 μm). Because >90% of the laser radiation was absorbed within the thermoplastic film, little direct heating of the tissue specimen by the laser occurred. The glass slide provided a large heat sink that confined the full-thickness transient focal melting of the thermoplastic material to the targeted region. The focally molten plastic wet the targeted tissue. After cooling and recrystallization. the film formed a local surface bond to the targeted tissue that was stronger than the adhesion forces of the tissue to the slide. The film and targeted cells were removed from the tissue specimen, resulting in focal microtransfer of the targeted tissue to the film surface.
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24. We are currently optimizing thinner, smoother transfer films and finer laser activation, which may allow for reproducible transfer of targeted single cells (R. F. Bonner et al., in preparation).
25. We thank S. K. Apple, J. P. Struewing, W. M. Linehan, C. D. Vocke, G. Oliver, A. Lash, M. J. Roth, M. J. Merino, P. H. Duray, I. A. Lubensky, L. V. Debelenko, V. E. Norman, and A. Coleman for tissue samples, technical support, or editing of the manuscript.