Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC–MS—A technical report
Laser capture microdissection (LCM) allows microscopic procurement of specific cell types from tissue sections. Here, we present an optimized workflow for coupling LCM to LC–MS/MS including: sectioning of tissue, a standard LCM workflow, protein digestion and advanced LC–MS/MS. Soluble proteins extr...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2016-03-01
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| Series: | EuPA Open Proteomics |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2212968515300246 |
| _version_ | 1811308590805286912 |
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| author | L. Staunton C. Tonry R. Lis S. Finn J. O’Leary M. Loda M. Bowden S.R. Pennington |
| author_facet | L. Staunton C. Tonry R. Lis S. Finn J. O’Leary M. Loda M. Bowden S.R. Pennington |
| author_sort | L. Staunton |
| collection | DOAJ |
| description | Laser capture microdissection (LCM) allows microscopic procurement of specific cell types from tissue sections. Here, we present an optimized workflow for coupling LCM to LC–MS/MS including: sectioning of tissue, a standard LCM workflow, protein digestion and advanced LC–MS/MS. Soluble proteins extracted from benign epithelial cells, their associated stroma, tumor epithelial cells and their associated stromal cells from a single patient tissue sample were digested and profiled using advanced LC–MS/MS. The correlation between technical replicates was R2 = 0.99 with a mean % CV of 9.55% ± 8.73. The correlation between sample replicates was R2 = 0.97 with a mean % CV of 13.83% ± 10.17. This represents a robust, systematic approach for profiling of the tumor microenvironment using LCM coupled to label-free LC–MS/MS. |
| first_indexed | 2024-04-13T09:27:06Z |
| format | Article |
| id | doaj.art-bf7aa297eabe4956b77a518f6631f6b5 |
| institution | Directory Open Access Journal |
| issn | 2212-9685 |
| language | English |
| last_indexed | 2024-04-13T09:27:06Z |
| publishDate | 2016-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | EuPA Open Proteomics |
| spelling | doaj.art-bf7aa297eabe4956b77a518f6631f6b52022-12-22T02:52:26ZengElsevierEuPA Open Proteomics2212-96852016-03-0110C192310.1016/j.euprot.2015.11.001Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC–MS—A technical reportL. Staunton0C. Tonry1R. Lis2S. Finn3J. O’Leary4M. Loda5M. Bowden6S.R. Pennington7Conway Institute, University College Dublin, Belfield, Dublin 4, IrelandConway Institute, University College Dublin, Belfield, Dublin 4, IrelandCenter for Molecular Oncologic Pathology, Dana-Farber Cancer Institute, 450 Brookline Ave., Boston, MA, USASt James’s Hospital, James’s St., Dublin 8, IrelandSt James’s Hospital, James’s St., Dublin 8, IrelandCenter for Molecular Oncologic Pathology, Dana-Farber Cancer Institute, 450 Brookline Ave., Boston, MA, USACenter for Molecular Oncologic Pathology, Dana-Farber Cancer Institute, 450 Brookline Ave., Boston, MA, USAConway Institute, University College Dublin, Belfield, Dublin 4, IrelandLaser capture microdissection (LCM) allows microscopic procurement of specific cell types from tissue sections. Here, we present an optimized workflow for coupling LCM to LC–MS/MS including: sectioning of tissue, a standard LCM workflow, protein digestion and advanced LC–MS/MS. Soluble proteins extracted from benign epithelial cells, their associated stroma, tumor epithelial cells and their associated stromal cells from a single patient tissue sample were digested and profiled using advanced LC–MS/MS. The correlation between technical replicates was R2 = 0.99 with a mean % CV of 9.55% ± 8.73. The correlation between sample replicates was R2 = 0.97 with a mean % CV of 13.83% ± 10.17. This represents a robust, systematic approach for profiling of the tumor microenvironment using LCM coupled to label-free LC–MS/MS.http://www.sciencedirect.com/science/article/pii/S2212968515300246Laser capture microdissectionLabel-free LC–MS/MS |
| spellingShingle | L. Staunton C. Tonry R. Lis S. Finn J. O’Leary M. Loda M. Bowden S.R. Pennington Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC–MS—A technical report EuPA Open Proteomics Laser capture microdissection Label-free LC–MS/ MS |
| title | Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC–MS—A technical report |
| title_full | Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC–MS—A technical report |
| title_fullStr | Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC–MS—A technical report |
| title_full_unstemmed | Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC–MS—A technical report |
| title_short | Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC–MS—A technical report |
| title_sort | profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to lc ms a technical report |
| topic | Laser capture microdissection Label-free LC–MS/ MS |
| url | http://www.sciencedirect.com/science/article/pii/S2212968515300246 |
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