scholarly journals Tissue preparation for laser capture microdissection and RNA extraction from fresh frozen breast tissue

BioTechniques ◽  
2007 ◽  
Vol 43 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Mary Morrogh ◽  
Narciso Olvera ◽  
Faina Bogomolniy ◽  
Patrick I. Borgen ◽  
Tari A. King
Keyword(s):  
Laser Capture Microdissection ◽  
Breast Tissue ◽  
Rna Extraction ◽  
Tissue Preparation ◽  
Fresh Frozen ◽  
Laser Capture

Detection of Cancer-Related Proteins in Fresh-Frozen Ovarian Cancer Samples Using Laser Capture Microdissection

2008 ◽  
pp. 35-45
Author(s):  
Dan-Arin Silasi ◽  
Ayesha B. Alvero ◽  
Jechiel Mor ◽  
Rui Chen ◽  
Han-Hsuan Fu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Laser Capture Microdissection ◽  
Fresh Frozen ◽  
Related Proteins ◽  
Laser Capture

Detection of Cancer-Related Proteins in Fresh-Frozen Ovarian Cancer Samples Using Laser Capture Microdissection

2008 ◽  
pp. 35-46
Author(s):  
Dan-Arin Silasi ◽  
Ayesha B. Alvero ◽  
Jechiel Mor ◽  
Rui Chen ◽  
Han-Hsuan Fu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Laser Capture Microdissection ◽  
Fresh Frozen ◽  
Related Proteins ◽  
Laser Capture

Application of Novel Laser Capture Microdissection and RT-PCR to a Functional Analysis of Osteopontin in Mouse Growth Plate Cartilage

2001 ◽  
Vol 7 (S2) ◽  
pp. 44-45
Author(s):  
R. Jacquet ◽  
J. Hillyer ◽  
J. Zhang ◽  
W. J. Landis
Keyword(s):  
Growth Plate ◽  
Laser Capture Microdissection ◽  
Critical Role ◽  
Rna Extraction ◽  
Normal Growth ◽  
Alternative Possibilities ◽  
Rt Pcr ◽  
Hypertrophic Growth ◽  
Growth Plate Cartilage ◽  
Laser Capture

The long bones of vertebrates, such as the tibiae and femurs of humans, extend in length by means of genotypic and phenotypic changes orchestrated by the chondrocytes comprising growth plate cartilage. Among the constituents synthesized by these cartilage cells, osteopontin (OPN), a phosphorylated glycoprotein, is thought to play a critical role in events leading to normal growth plate function and ultimate mineralization of the deeper zones of this cartilage region. The precise role of OPN, however, is uncertain with regard to mineralization, and present evidence supports the alternative possibilities that the protein may be either facilitative or inhibitory to mineral deposition. in order to investigate OPN function in a model growth plate, cartilage from normal 1-11 day old postnatal mice was examined by the novel techique of laser capture microdissection (LCM) followed by RT-PCR to obtain a measure of OPN gene expression by chondrocytes of known age and specific location in the plate. LCM permits identification of individual or clusters of cells within a tissue section and subsequent unique isolation (“capture”) of such cells for a variety of molecular analyses.In this study, mouse tibiae were dissected, placed in RNAlater (Ambion, Austin, TX) to preserve message, and stored at −20°C. Sections (5 μm thick) of fresh frozen developing epiphyseal growth plates were obtained in a cryostat maintained at −20°C, fixed briefly in 70% ethanol, and stained with eosin. Sections were examined in a Pixcell LCM system (Arcturus Engineering, Mountain View, CA) where chondrocytes were attached to the surface of polymer film substrates and lifted free of sections. in separate experiments, ∼200-1200 cells were captured and analyzed. Substrates were transferred to Eppendorf tubes containing RNA extraction buffer. RNA was extracted from cells by microisolation (Stratagene, La Jolla, CA), DNAse-treated, reverse-transcribed, and then subjected to PCR (40 cycles) with AmpliTaq DNA polymerase (PE Applied Biosystems, Foster City, CA). Ethidium bromide agarose gels revealed OPN mRNA from groups of chondrocytes isolated from whole cartilage and from resting, proliferating, and hypertrophic growth plate zones from the mouse tibiae. Brain cells captured by LCM from the same mouse sections served as positive controls and reactions containing no reverse transcriptase were negative controls. 18S rRNA was used as a marker for semiquantitation and standardization of expressed message from captured cells.

scholarly journals Laser Capture Microdissection of Uredinia Formed by Melampsora larici-populina Revealed a Transcriptional Switch Between Biotrophy and Sporulation

2010 ◽  
Vol 23 (10) ◽  
pp. 1275-1286 ◽  
Author(s):  
Stéphane Hacquard ◽  
Christine Delaruelle ◽  
Valérie Legué ◽  
Emilie Tisserant ◽  
Annegret Kohler ◽  
...  
Keyword(s):  
Laser Capture Microdissection ◽  
Molecular Mechanisms ◽  
Rust Fungi ◽  
Tissue Preparation ◽  
Cell Defense ◽  
Genome Wide ◽  
Study Gene Expression ◽  
Transcriptional Switch ◽  
Host Infection ◽  
Laser Capture

The foliar rust caused by the basidiomycete Melampsora larici-populina is the main disease affecting poplar plantations in Europe. The biotrophic status of rust fungi is a major limitation to study gene expression of cell or tissue types during host infection. At the uredinial stage, infected poplar leaves contain distinct rust tissues such as haustoria, infection hyphae, and uredinia with sporogenous hyphae and newly formed asexual urediniospores. Laser capture microdissection (LCM) was used to isolate three areas corresponding to uredinia and subjacent zones in the host mesophyll for expression analysis with M. larici-populina whole-genome exon oligoarrays. Optimization of tissue preparation prior to LCM allowed isolation of RNA of good integrity for genome-wide expression profiling. Our results indicate that the poplar rust uredinial stage is marked by distinct genetic programs related to biotrophy in the host palisade mesophyll and to sporulation in the uredinium. A strong induction of transcripts encoding small secreted proteins, likely containing rust effectors, is observed in the mesophyll, suggesting a late maintenance of suppression of host defense in the tissue containing haustoria and infection hyphae. On the other hand, cell cycle and cell defense rescue transcripts are strongly accumulated in the sporulation area. This combined LCM-transcriptomic approach brings new insights on the molecular mechanisms underlying urediniospore formation in rust fungi.

Fixation and Laser Capture Microdissection of Plant Tissue for RNA Extraction and RNASeq Library Preparation

2018 ◽  
Vol 3 (1) ◽  
pp. 14-32
Author(s):  
Suchitra Chavan ◽  
Elise Schnabel ◽  
Christopher Saski ◽  
Julia Frugoli
Keyword(s):  
Laser Capture Microdissection ◽  
Plant Tissue ◽  
Rna Extraction ◽  
Library Preparation ◽  
Laser Capture

A Laser Capture Microdissection Protocol That Yields High Quality RNA from Fresh-frozen Mouse Bones

10.3791/60197 ◽  
2019 ◽  
Author(s):  
Ana Marek ◽  
Christiane Schüler ◽  
María Satué ◽  
Barbara Haigl ◽  
Reinhold G. Erben
Keyword(s):  
Laser Capture Microdissection ◽  
High Quality ◽  
Fresh Frozen ◽  
Laser Capture
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