Abbreviations:EVA (ethylene-vinyl acetate), FFPE (formalin-fixed paraffin-embedded), IHC (immunohistochemical), IR-LCM (infrared LCM), LCM (laser capture microdissection), UV-LCM (ultraviolet LCM)
- •Recognize the newest techniques in biomedical research.
- •Describe how these techniques can be utilized and their limitations.
- •Describe the potential impact of these techniques.
What LCM Does
- •LCM is a technique that isolates cells of interest or even a single cell from a heterogeneous tissue specimen using a laser source under microscopic visualization.
- •Cells isolated by LCM contain intact DNA, RNA, and proteins for downstream molecular analysis.
- •LCM is capable of isolating diseased cells from the primary lesion without altering their molecular signatures.
- •LCM can be applied to a wide variety of tissue and cellular preparations.
- •In the absence of a cover slip, the optical resolution of complex tissues may be limited.
- •The reliance on visual identification of target cells creates room for human error.
- •Unlike IR-LCM, UV-LCM is limited by the potential to induce UV damage in the circumferential cells, which may be subsequently collected for analysis.
Principles of LCM
Advantages and Limitations
LCM in Cutaneous Research
Summary and Future Directions
Conflict of Interest
Multiple Choice Questions
- 1.What does LCM do?
- A.Sorts cells based on morphology (size, granularity, density)
- B.Sorts cells with either IR or UV laser technology
- C.Collects cells of interest with laser technology
- D.Photoablates cells of interest with laser technology
- 2.What is the thinnest diameter of the UV-LCM laser beam?
- A.0.5 μm
- B.5.0 μm
- C.7.5 μm
- D.30 μm
- 3.Which technique uses a thermosensitive EVA film to sequester cells of interest?
- C.Laser microbeam microdissection
- 4.In the absence of a cover slip, the optical resolution of complex tissues may be limited. This issue can be addressed by:
- A.Using a temporary cover slip
- B.Increasing the thickness of the tissue section
- C.Decreasing the thickness of the tissue section
- D.Staining with immunohistochemistry
- 5.Which of the following statements regarding the LCM technique is NOT true?
- A.UV-LCM is better suited for single cell microdissection
- B.IR-LCM is more time consuming than UV-LCM
- C.The EVA membrane undergoes conformational change when exposed to IR laser energy
- D.The downstream analysis of proteins is limited in FFPE tissue sections due to undesirable protein and nucleic acid crosslinking
- Quiz and brief explanation of correct answers
- Teaching Slides
- Laser capture microdissection.Science. 1996; 274: 998-1001
- Laser-capture microdissection.Nat Protoc. 2006; 1: 586-603
- Immuno-LCM: laser capture microdissection of immunostained frozen sections for mRNA analysis.Am J Pathol. 1999; 154: 61-66
- Laser capture microdissection in pathology.J Clin Pathol. 2000; 53: 666-672
- Narrow band ultraviolet B treatment for human vitiligo is associated with proliferation, migration, and differentiation of melanocyte precursors.J Invest Dermatol. 2015; 135: 2068-2076
- In vitro cancer cell-ECM interactions inform in vivo cancer treatment.Adv Drug Deliv Rev. 2016; 97: 270-279
- Age-associated increase in skin fibroblast-derived prostaglandin E2 contributes to reduced collagen levels in elderly human skin.J Invest Dermatol. 2015; 135: 2181-2188
- Laser capture microdissection in the tissue biorepository.J Biomol Tech. 2010; 21: 120-125
- Laser capture microdissection for the investigative pathologist.Vet Pathol. 2014; 51: 257-269
- Gene expression differences predict treatment outcome of Merkel cell carcinoma patients.J Skin Cancer. 2014; 2014: 596459
- Identification of expressed genes by laser-mediated manipulation of single cells.Nat Biotechnol. 1998; 16: 737-742
- A novel hypoxia-associated subset of FN1 high MITF low melanoma cells: identification, characterization, and prognostic value.Mod Pathol. 2014; 27: 1088-1100
- Microarray analysis of fluoro-gold labeled rat dopamine neurons harvested by laser capture microdissection.J Neurosci Methods. 2005; 143: 95-106
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