mRNA extraction and subsequent RT-polymerase chain reaction (PCR)-based expression analysis from laser-microdissected material is by now a well-established and reproducible method. Most routinely stored tissue samples are preserved as formalin-fixed, paraffin-embedded materials. While this allows for a convenient storage and stable preservation of nucleic acids, deparaffinization before staining for laser microdissection may result in a significant loss of mRNA quality and consequently of PCR sensitivity. We describe a method of isolating anatomic compartments from non-deparaffinized, formalin-fixed, and paraffin-embedded tissues by laser-assisted microdissection which allows for a highly efficient mRNA retrieval.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-61779-163-5_5 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Differential somatic coding variant landscapes between laser microdissected luminal epithelial cells from canine mammary invasive ductal solid carcinoma and comedocarcinoma.
BMC Cancer
December 2024
Department of Veterinary Biosciences, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland.
Background: Breast cancer (BC) is the most common cancer in women. Likewise, canine mammary tumors (CMT) represent the most common cancer in intact female dogs and develop in the majority spontaneously. Similarities exist in clinical presentation, histopathology, biomarkers, and treatment.
View Article and Find Full Text PDFA framework for ultra-low-input spatial tissue proteomics.
Cell Syst
November 2023
Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), Spatial Proteomics Group, Berlin, Germany. Electronic address:
Spatial proteomics combining microscopy-based cell phenotyping with ultrasensitive mass-spectrometry-based proteomics is an emerging and powerful concept to study cell function and heterogeneity in (patho)physiology. However, optimized workflows that preserve morphological information for phenotype discovery and maximize proteome coverage of few or even single cells from laser microdissected tissue are currently lacking. Here, we report a robust and scalable workflow for the proteomic analysis of ultra-low-input archival material.
View Article and Find Full Text PDFMass Spectrometry-Based Analysis of Histone Posttranslational Modifications from Laser Microdissected Samples.
Methods Mol Biol
September 2023
Department of Experimental Oncology, IEO, European Institute of Oncology IRCCS, Milan, Italy.
The analysis of histone posttranslational modifications (PTMs) in clinical samples has gained considerable interest due to the increasing knowledge about the implication of epigenetics in a multitude of physiological and pathological processes. Mass spectrometry (MS) has emerged as the most accurate and versatile tool to detect and quantify histone PTMs and has also been applied to clinical specimens, thanks to protocols developed during the past years. However, the requirement for relatively large amounts of material has so far impaired the application of these approaches to samples available in limited amounts.
View Article and Find Full Text PDFDisease-related protein co-expression networks are associated with the prognosis of resectable node-positive pancreatic ductal adenocarcinoma.
Sci Rep
August 2022
Department of Surgery, National Hospital Organization Sendai Medical Center, Sendai, Miyagi, 983-8520, Japan.
Pancreatic ductal adenocarcinoma (PDAC) is a multifactorial disease, the molecular profile of which remains unclear. This study aimed at unveiling the disease-related protein networks associated with different outcomes of resectable, node-positive PDAC cases. We assessed laser-microdissected cancerous cells from PDAC tissues of a poor outcome group (POG; n = 4) and a better outcome group (BOG; n = 4).
View Article and Find Full Text PDFSpatial epi-proteomics enabled by histone post-translational modification analysis from low-abundance clinical samples.
Clin Epigenetics
July 2021
Department of Experimental Oncology, IEO, European Institute of Oncology IRCCS, Milan, Italy.
Article Synopsis
- This study investigates how to analyze histone post-translational modifications (PTMs) from small clinical samples, specifically targeting breast cancer tissues, to enhance our understanding of epigenetic mechanisms in diseases.
- Researchers developed a streamlined method that allows for the analysis of histone PTMs using as few as 1000 cells, significantly reducing the sample size needed compared to previous methods.
- The successful application of this method in identifying histone mark differences in tumor regions suggests potential for new approaches in spatial epi-proteomics, which could lead to discovering new epigenetic biomarkers and understanding tumor heterogeneity better.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!