Project description:The different stages of the optic fissure can be clearly visualized by making sagittal sections through the mouse eye during early development which represent the optic fissure at open (E10.5), closing (E11.5) and fused (E12.5) states. Laser capture microdissection (LCM) was employed to dissect tissue from the margins of the optic fissure consisting of the outer (presumptive RPE) and inner (presumptive neurosensory retina) layers of the retina. An approximately square-shaped block of optic fissure (50 x 50 mm) was dissected from each side of the fissure. Two rounds of linear amplification were performed on RNA isolated from each of the samples prior to microarray hybridization. Expression data were gathered in biological triplicate at E10.5, E11.5 and duplicate at E12.5, each array representing pooled optic fissure tissue from three embryos from a single litter. Expression signals were ascertained from 45,101 probe sets and normalized across arrays. Experiment Overall Design: Differential gene expression over the course of three days in laser capture microdissected tissue from wild type mouse embryos.
Project description:Liquid chromatography mass spectrometry was used to study tumor matrisome from laser capture microdissected FFPE tissue sections of human neck and squamous cell carcinoma (HnSCC) xenograft grown in mouse.
Project description:The objective of this study was to investigate how active dermal connective tissue composition dynamics affect long term protein expression in the epidermis. After surgically removing damaged skin to its full thickness at the site of a deep burn in ten patients, we randomized three adjacent test areas within the wounds to receive either: 1) no dermal component as control, 2) a permanent artificial dermal matrix or 3) a temporary dressing to induce formation of granulation tissue. Each area was then covered with an autologous superficial healty skin transplant as the epidermal indicator. One year later, biopsy samples were taken from each site. The epidermis and dermis were selectively microdissected by laser-capture from cut tissue biopsy sections, and analyzed using untargeted non-labelled quantitative proteomics.
Project description:Frozen blocks of human embryonic limb tissue were sectioned and laser capture microdissected to collect three human chodrocyte tissues - precursor, differentiated and hypertrophic chondrocytes.Total RNA including miRNAs was isolated and analyzed using Applied Biosystems OpenArray® Real-Time PCR system.
Project description:Microarray data from this study represent the first global transcriptional survey of gene expression during early compared to late diaphragm formation. Based on the assumption that Congenital diaphragmatic hernia (CDH) candidate genes will have similar temporal expression trends and will be part of the same biological pathways as those already implicated in CDH, we used known CDH-associated genes as “baits” to filter expression microarray data sets (GSEA and STEM) to identify a list of novel CDH candidate genes. Laser-capture microdissected C57BL/6J mouse embryonic diaphragm tissues were studied at three key time-points during normal development. We studied the primordial diaphragm’s pleuroperitoneal folds at E11.5 and E12.5, and the muscularized mature diaphragm at E16.5 to identify genes and pathways that characterize important stages in diaphragm morphogenesis.
Project description:We combined RNA sequencing of laser capture microdissected-tissues with single molecule transcript imaging to obtain a spatial gene expression map of the ILF and its associated FAE in the mouse small intestine.
Project description:E12.5 wild-type and NestinCre Foxa1/2 Flox/ Flox mutant embryos were quickly snap-frozen, then 10um-thick cryostat coronal sections of the midbrain were cut and mounted on membrane slides (Zeiss) and the floor plate was Laser Capture Microdissected. RNA was extracted using the Picopure RNA Isolation Kit RNA sequencing library was prepared using the Ovation RNA-Seq system (Nugen). RNA-Seq libraries were sequenced on the Illumina GAIIx.
Project description:The purpose of this study was to identify differentially expressed genes in laser-capture microdissected (LCM) invasive mammary carcinomas (IMCs).