ArticleJournal of neuroscience methods2012
Rapid expression profiling of brain microvascular endothelial cells by immuno-laser capture microdissection coupled to TaqMan(®) low density array.
Article in Journal of neuroscience methods, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
10 citing papers in PubMed.
- Immuno-laser capture microdissection of perfusion-fixed mouse brain tissue coupled to RNA-seq.Journal of neuroscience methods · 2025Article
- Analysis of neuroendocrine clones in NSCLCs using an immuno-guided laser-capture microdissection-based approach.Cell reports methods · 2022Article
- Proteome of the Luminal Surface of the Blood-Brain Barrier.Proteomes · 2021Review
- Human ES-derived MSCs correct TNF-α-mediated alterations in a blood-brain barrier model.Fluids and barriers of the CNS · 2019Article
- Altered Expression of Genes Encoding Neurotransmitter Receptors in GnRH Neurons of Proestrous Mice.Frontiers in cellular neuroscience · 2016Article
- In vitro model of the blood-brain barrier established by co-culture of primary cerebral microvascular endothelial and astrocyte cells.Neural regeneration research · 2015Article
- Transcript analysis of laser capture microdissected white matter astrocytes and higher phenol sulfotransferase 1A1 expression during autoimmune neuroinflammation.Journal of neuroinflammation · 2015Article
- Identification of ATP8B1 as a blood-brain barrier-enriched protein.Cellular and molecular neurobiology · 2014Article
- Field evaluation of TaqMan Array Card (TAC) for the simultaneous detection of multiple respiratory viruses in children with acute respiratory infection.Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology · 2013Article
- Active induction of experimental autoimmune encephalomyelitis by MOG35-55 peptide immunization is associated with differential responses in separate compartments of the choroid plexus.Fluids and barriers of the CNS · 2012Article
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Authors and funding
4 authors.
Funding
Abstract
Immuno-laser capture microdissection (immuno-LCM) enables highly selective retrieval of designated cell populations from their in situ locations in complex tissue like the brain. However, the amount of tissue acquired by immuno-LCM is extremely limited, and the RNA purification, amplification and labeling steps necessary for expression analysis by hybridization microarray are tedious and time consuming. This report therefore describes a protocol in which these RNA steps are eliminated altogether, yet allows for global gene profiling. Specifically, immuno-LCM tissue was solubilized and the extract directly subjected to reverse transcription to generate cDNA. Pre-amplification of cDNA was performed next, and then relative expression of 96 different immune-related genes simultaneously determined by quantitative real-time PCR using a microfluidic card TaqMan(®) Low Density Array (TLDA). This protocol was highly reproducible and extremely sensitive, demonstrating high correlation of raw Ct values among both technical and biological replicate samples when using only 1/32 of total pre-amplified cDNA obtained from as little as 500 LCM 'shots.' As this abridged protocol takes only approximately 7h from LCM tissue acquisition to analysis by TLDA, it can prove a very effective tool for both screening and validation purposes when investigating gene regulation in health and disease of the nervous system and other tissues.
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