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MicroRNA Profiling Using RNA from Microdissected Immunostained Tissue

mRNA expression profiling has been used to define molecular subtypes of human breast cancer. Also microRNAs have been investigated in these breast cancer subtypes. However, little is known regarding the microRNA signature of healthy luminal and basal breast epithelial cells. Theref ...

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Profiling Solid Tumor Heterogeneity by LCM and Biological MS of Fresh-Frozen Tissue Sections

The heterogeneous nature of solid tumors represents a common problem in mass spectrometry (MS)-based analysis of fresh-frozen tissue specimens. Here, we describe a method that relies on synergy between laser capture microdissection (LCM) and MS for enhanced molecular profiling of so ...

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Amplification Testing in Breast Cancer by Multiplex Ligation-Dependent Probe Amplification of Microdissected Tissue

This chapter describes a method for the rapid assessment of gene copy numbers in laser-microdissected materials using multiplex ligation-dependent probe amplification (MLPA). An MLPA is a powerful multiplex PCR technique that can identify gains, amplification, or losses of up to 50 ge ...

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Probing the CNS Microvascular Endothelium by Immune-Guided Laser-Capture Microdissection Coupled to Quantitative RT-PCR

Laser-capture microdissection (LCM) allows for retrieval of distinct populations of cells from their closely surrounding neighbors in situ. As such, LCM is highly advantageous for investigating gene expression along the central nervous system (CNS) microvascular endotheli ...

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Laser-Capture Microdissection for Factor VIII-Expressing Endothelial Cells in Cancer Tissues

The immunostaining based Laser-capture microdissection (LCM) method called immune-LCM allows us to quantify the mRNA. Immune-LCM has recently been introduced to enhance identification of cells carrying a particular protein from frozen tissue samples. We have recently perform ...

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Laser-Capture Microdissection and Analysis of Liver Endothelial Cells from Patients with BuddChiari Syndrome

Myeloproliferative neoplasms (MPN) are clonal hematological malignancies that are frequently �associated with an acquired somatic mutation in JAK2 (JAK2V617F). Patients with MPN are at a high risk of developing thrombotic events. Endothelial cell (EC) abnormalities are thou ...

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Laser-Capture Microdissection of Hyperlipidemic/ApoE/ Mouse Aorta Atherosclerosis

Atherosclerosis is a transmural chronic inflammatory condition of small and large arteries that is associated with adaptive immune responses at all disease stages. However, impacts of adaptive immune reactions on clinically apparent atherosclerosis such as intima lesion (pla ...

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Specific RNA Collection from the Rat Endolymphatic Sac by Laser-Capture Microdissection (LCM): LCM of a Very Small Organ Surrounded by Bony Tissues

Laser-capture microdissection (LCM) is an excellent tool to selectively obtain target tissue or cells. The endolymphatic sac (ES) is part of the inner ear, and a large part of the ES is located in the temporal bone. The rat ES is conventionally harvested using stereomicroscopy. In this method, conta ...

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Gene Expression Profiling in Laser-Microdissected Bone Marrow Megakaryocytes

Gene expression analysis of the megakaryocytic lineage requires isolation of megakaryocytes from their bone marrow microenvironment. Laser microdissection of megakaryocytes from diagnostic bone marrow biopsies allows analysis of standardised formalin-fixed samp ...

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Laser-Capture Microdissection of Developing Barley Seeds and cDNA Array Analysis of Selected Tissues

Laser microdissection provides a useful method for isolating specific cell types from complex biological samples for downstream applications. In contrast to the texture of mammalian cells, most plant tissues exhibit a cell organization with hard, cellulose-containing cell wal ...

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The Use of Laser Capture Microdissection on Adult Human Articular Cartilage for Gene Expression Analysis

The integrity of articular cartilage is maintained by chondrocytes, the sole type of cell that resides within the tissue. The noncalcified region of articular cartilage can be divided into three zones based on histological features, in which the chondrocyte metabolism is known to differ o ...

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Gene Expression Using the PALM System

The PALM Robot MicroBeam laser microdissection system can isolate specified cells from complex tissues section, in a rapid and precise manner. Combined with other methods, PALM may be used for gene expression elucidating the role of specialized cell type in physiological and pathologi ...

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Quantitative RT-PCR Gene Expression Analysis of a Laser Microdissected Placenta: An Approach to Study Preeclampsia

Preeclampsia is still one of the leading causes of maternal and neonatal mortality and morbidity. Despite intensive research, the cause of preeclampsia has not yet been established. However, for a variety of reasons, the numerous aspects of normal and pathological pregnancies, particu ...

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Immunoguided Microdissection Techniques

Over the past 15 years, laser-based microdissection has improved the precision by which scientists can procure cells of interest from a heterogeneous tissue section. However, for studies that require a large amount of material (e.g., proteomics) or for cells that are scattered and difficult ...

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Optimized RNA Extraction from Non-deparaffinized, Laser-Microdissected Material

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-embe ...

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Detection and Quantification of MicroRNAs in Laser-Microdissected Formalin-Fixed Paraffin-Embedded Breast Cancer Tissues

MicroRNAs (miRNAs) are a class of small endogenous non-coding RNAs that regulate gene expression post-transcriptionally through targeting protein-coding mRNAs for cleavage or translational repression, and thus play key roles in cellular fate-determinant pathways. Both pr ...

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Laser Capture Microdissection Applications in Breast Cancer Proteomics

Breast cancer tissues are characterized by cellular heterogeneity, representing a mixture of, e.g., healthy epithelial ducts, invasive or in situ tumor cells, surrounding stroma, infiltrating immune cells, blood vessels, and capillaries. As a consequence, protein extracts from wh ...

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Proteomic Analysis of Laser Microdissected Ovarian Cancer Tissue with SELDI-TOF MS

The evolution of ovarian cancer will highly depend on platinum sensitivity or resistance. In an attempt to better understand this mechanism of resistance, we combined laser microdissection (LMD) of ovarian tumor cells with mass spectrometric techniques. To obtain disease-specif ...

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LCM Assisted Biomarker Discovery from Archival Neoplastic Gastrointestinal Tissues

Expression array analysis of epithelial mRNA to identify biomarkers of premalignant and malignant conditions in the gastrointestinal (GI) tract is an area of intense study. Archived formalin-fixed paraffin-embedded (FFPE) tissues documenting these changes are readily avai ...

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Application of Laser Microdissection and Quantitative PCR to Assess the Response of Esophageal Cancer to Neoadjuvant Chemo-Radiotherapy

Tissues are complicated three-dimensional structures, composed of different types of interacting cells. Since the cell population of interest might constitute only a minor fraction of the total tissue volume, the problem of tissue heterogeneity has been a major barrier to the molecul ...

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