Laser capture microdissection technology

Espina, Virginia; Heiby, Michael; Pierobon, Mariaelena; Liotta, Lance A.

doi:10.1586/14737159.7.5.647

Cited by 166 publications

(96 citation statements)

References 93 publications

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“…Over the last few years, advances in protein extraction methodologies have made it possible to retrieve invaluable proteomic data from FFPE materials using MS (40,43,44). The application of pre-digestion enrichment processes, such as LCM, ensures that downstream proteomic analyses reflect predominantly the cells of interest, and also improves the detection, coverage and reproducibility of peptide measurements (45).…”

Section: Discussionmentioning

confidence: 99%

In Silico Analysis Validates Proteomic Findings of Formalin-fixed Paraffin Embedded Cutaneous Squamous Cell Carcinoma Tissue

Azimi

Kaufman

Ali

et al. 2016

CGP

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Section: Discussionmentioning

confidence: 99%

In Silico Analysis Validates Proteomic Findings of Formalin-fixed Paraffin Embedded Cutaneous Squamous Cell Carcinoma Tissue

Azimi

Kaufman

Ali

et al. 2016

CGP

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“…This is expected to alter patterns of gene expression compared with cells in vivo. Therefore, we used laser-capture microdissection (Espina et al, 2007) combined with immunostaining for cellular recognition. As standard immunohistochemistry protocols lower RNA quality (Fend et al, 1999), we optimised a protocol to (Pascal et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

TEAD1 and c-Cbl are novel prostate basal cell markers that correlate with poor clinical outcome in prostate cancer

et al. 2008

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Prostate cancer is the most frequently diagnosed male cancer, and its clinical outcome is difficult to predict. The disease may involve the inappropriate expression of genes that normally control the proliferation of epithelial cells in the basal layer and their differentiation into luminal cells. Our aim was to identify novel basal cell markers and assess their prognostic and functional significance in prostate cancer. RNA from basal and luminal cells isolated from benign tissue by immunoguided laser-capture microdissection was subjected to expression profiling. We identified 112 and 267 genes defining basal and luminal populations, respectively. The transcription factor TEAD1 and the ubiquitin ligase c-Cbl were identified as novel basal cell markers. Knockdown of either marker using siRNA in prostate cell lines led to decreased cell growth in PC3 and disrupted acinar formation in a 3D culture system of RWPE1. Analyses of prostate cancer tissue microarray staining established that increased protein levels of either marker were associated with decreased patient survival independent of other clinicopathological metrics. These data are consistent with basal features impacting on the development and clinical course of prostate cancers.

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“…Although such selection is possible by laser-capture microdissection (LCM), successful use of this technology has so far overwhelmingly been in the areas of RNA and DNA assays. In the few cases where LCM was combined with MS analysis, many thousands cells were collected (taking presumably several hours or days of selection), to obtain the protein amounts typical for MS (6). Only a few investigators have attempted to measure smaller cell numbers.…”

Section: Ass Spectrometry (Ms) Is Inherently An Extremely Sensitivementioning

confidence: 99%

Quantitative proteomic analysis of single pancreatic islets

Waanders

Chwalek

Monetti

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

201

213

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Technological developments make mass spectrometry (MS)-based proteomics a central pillar of biochemical research. MS has been very successful in cell culture systems, where sample amounts are not limiting. To extend its capabilities to extremely small, physiologically distinct cell types isolated from tissue, we developed a high sensitivity chromatographic system that measures nanogram protein mixtures for 8 h with very high resolution. This technology is based on splitting gradient effluents into a capture capillary and provides an inherent technical replicate. In a single analysis, this allowed us to characterize kidney glomeruli isolated by laser capture microdissection to a depth of more than 2,400 proteins. From pooled pancreatic islets of Langerhans, another type of ''miniorgan,'' we obtained an in-depth proteome of 6,873 proteins, many of them involved in diabetes. We quantitatively compared the proteome of single islets, containing 2,000 -4,000 cells, treated with high or low glucose levels, and covered most of the characteristic functions of beta cells. Our ultrasensitive analysis recapitulated known hyperglycemic changes but we also find components up-regulated such as the mitochondrial stress regulator Park7. Direct proteomic analysis of functionally distinct cellular structures opens up perspectives in physiology and pathology.hyperglycemia ͉ liquid chromatography-mass spectrometry ͉ quantitative proteomics ͉ replay technology

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Laser capture microdissection technology

Cited by 166 publications

References 93 publications

In Silico Analysis Validates Proteomic Findings of Formalin-fixed Paraffin Embedded Cutaneous Squamous Cell Carcinoma Tissue

In Silico Analysis Validates Proteomic Findings of Formalin-fixed Paraffin Embedded Cutaneous Squamous Cell Carcinoma Tissue

TEAD1 and c-Cbl are novel prostate basal cell markers that correlate with poor clinical outcome in prostate cancer

Quantitative proteomic analysis of single pancreatic islets

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