Identification of Vascular Breast Tumor Markers by Laser Capture Microdissection and Label-Free LC−MS

2011 ◽  
Vol 10 (5) ◽  
pp. 2479-2493 ◽  
Author(s):  
Jennifer J. Hill ◽  
Tammy-Lynn Tremblay ◽  
Ally Pen ◽  
Jie Li ◽  
Anna C. Robotham ◽  
...  
Keyword(s):  
Tumor Markers ◽  
Laser Capture Microdissection ◽  
Breast Tumor ◽  
Label Free ◽  
Laser Capture

scholarly journals Label-Free Comparative Proteomic Analysis Combined with Laser-Capture Microdissection Suggests Important Roles of Stress Responses in the Black Layer of Maize Kernels

2020 ◽  
Vol 21 (4) ◽  
pp. 1369
Author(s):  
Quanquan Chen ◽  
Ran Huang ◽  
Zhenxiang Xu ◽  
Yaxin Zhang ◽  
Li Li ◽  
...  
Keyword(s):  
Laser Capture Microdissection ◽  
Stress Responses ◽  
Label Free ◽  
Kernel Development ◽  
Flight Mass Spectrometry ◽  
Specific Proteins ◽  
Response To Stress ◽  
Black Layer ◽  
Maize Kernels ◽  
Laser Capture

The black layer (BL) is traditionally used as an indicator for kernel harvesting in maize, as it turns visibly dark when the kernel reaches physiological maturity. However, the molecular roles of BL in kernel development have not been fully elucidated. In this work, microscopy images showed that BL began to appear at a growth stage earlier than 10 days after pollination (DAP), and its color gradually deepened to become dark as the development period progressed. Scanning electron microscopy observations revealed that BL is a tissue structure composed of several layers of cells that are gradually squeezed and compressed during kernel development. Laser-capture microdissection (LCM) was used to sample BL and its neighboring inner tissue, basal endosperm transfer layer (BETL), and outer tissue, inner epidermis (IEP), from 20 DAP of kernels. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry profiling (MALDI-TOF MS profiling) detected 41, 104, and 120 proteins from LCM-sampled BL, BETL, and IEP, respectively. Gene ontology (GO) analysis indicated that the 41 BL proteins were primarily involved in the response to stress and stimuli. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis found that the BL proteins were enriched in several defense pathways, such as the ascorbate and aldarate metabolic pathways. Among the 41 BL proteins, six were BL-specific proteins that were only detected from BL. Annotations of five BL-specific proteins were related to stress responses. During kernel development, transcriptional expression of most BL proteins showed an increase, followed by a decrease, and reached a maximum zero to 20 DAP. These results suggest a role for BL in stress responses for protecting filial tissue against threats from maternal sides, which helps to elucidate the biological functions of BL.

scholarly journals Laser Capture Microdissection-Assisted Protein Biomarker Discovery from Coccidioides-Infected Lung Tissue

2020 ◽  
Vol 6 (4) ◽  
pp. 365
Author(s):  
Natalie M. Mitchell ◽  
Surendra Dasari ◽  
Thomas E. Grys ◽  
Douglas F. Lake
Keyword(s):  
Laser Capture Microdissection ◽  
Biomarker Discovery ◽  
Pathogenic Fungus ◽  
Dependent Manner ◽  
Label Free ◽  
Human Lungs ◽  
Human Proteins ◽  
Laser Capture

Laser capture microdissection (LCM) coupled to label-free quantitative mass spectrometry is a viable strategy to identify biomarkers from infected tissues. In this study, LCM was employed to take a “snapshot” of proteins produced in vivo during Coccidiodies spp. infection in human lungs. Proteomic analysis of LCM lung sections revealed hundreds of hosts and Coccidioidal proteins. Twenty-seven highly abundant Coccidioides spp. proteins were identified which do not share significant sequence orthology with human proteins. Three of the 27 Coccidioidal proteins are also potential Coccidoides-specific biomarkers, as they also do not share sequence homology to any other pathogenic fungus or microbe. Gene ontology analysis of the 27 biomarker candidate proteins revealed enriched hydrolase activity and increased purine and carbohydrate metabolism functions. Finally, we provide proteomic evidence that all 27 biomarker candidates are produced by the fungus when grown in vitro in a media- and growth-phase dependent manner.

scholarly journals A quantitative proteomic workflow for characterization of frozen clinical biopsies: Laser capture microdissection coupled with label-free mass spectrometry

2012 ◽  
Vol 77 ◽  
pp. 433-440 ◽  
Author(s):  
John P. Shapiro ◽  
Sabyasachi Biswas ◽  
Anand S. Merchant ◽  
Anjali Satoskar ◽  
Cenny Taslim ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Laser Capture Microdissection ◽  
Label Free ◽  
Laser Capture

Isolating Human Notochord Using Laser Capture Microdissection

2016 ◽  
Vol 77 (S 02) ◽  
Author(s):  
Youssef Yakkioui ◽  
Remco Santegoeds ◽  
Koo van Overbeeke ◽  
Andreas Herrler ◽  
Yasin Temel
Keyword(s):  
Laser Capture Microdissection ◽  
Laser Capture

scholarly journals Stromal MED12 exon 2 mutations in complex fibroadenomas of the breast

2020 ◽  
pp. jclinpath-2020-207062
Author(s):  
Edaise M da Silva ◽  
Francisco Beca ◽  
Ana Paula Martins Sebastiao ◽  
Melissa P Murray ◽  
Catarina Silveira ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Laser Capture Microdissection ◽  
Sanger Sequencing ◽  
Mediator Complex ◽  
Telomerase Reverse Transcriptase ◽  
Exon 2 ◽  
Stromal Component ◽  
Hotspot Mutations ◽  
Laser Capture

AimsHere we explore the presence of mediator complex subunit 12 (MED12) exon 2 and telomerase reverse transcriptase (TERT) promoter hotspot mutations in complex fibroadenomas (CFAs) of the breast.MethodsThe stromal components from 18 CFAs were subjected to Sanger sequencing of MED12 exon 2 and the TERT promoter hotspot loci. The epithelial and stromal components of two MED12 mutated CFAs were subjected to laser capture microdissection, and Sanger sequencing of MED12 exon 2, TERT promoter and PIK3CA exons 9 and 20, separately.ResultsMED12 exon 2 mutations were identified in the stroma of 17% of CFAs. The analyses of epithelial and stromal components, microdissected separately, revealed that MED12 mutations were restricted to the stroma. No TERT promoter or PIK3CA mutations in exons 9 and 20 were detected in analysed CFAs.ConclusionsLike conventional fibroadenomas, MED12 exon 2 mutations appear to be restricted to the stromal component of CFAs, supporting the notion that CFAs are stromal neoplasms.

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