microdissected Protocols

Category: Genetics and Genomics: Cancer Genetics Protocols

Investigators can utilize X chromosome inactivation (methylation) to determine the clonality status of a tumor or premalignant lesion in females. The technique is based on a methylation-sensitive restriction enzyme and analysis of a polymorphic locus on the X chromosome. Clonal cell populations will show "loss" of the non-methylated allele after restriction digest. The assay can be performed on DNA recovered from microdissected samples. Both frozen tissue and fixed-embedded tissue can be used. - [Read Clonality - X Chromosome Inactivation Assay Protocol]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Three Ambion kits were used to quantitate specific miRNAs and to detect differential miRNA expression in various mouse brain regions and cell types isolated by laser capture microdissection (LCM). These techniques can be applied to studying miRNA in other species, tissues, and cell types. Includes: Obtain Laser Capture Microdissected Samples; Isolate miRNA from LCM Samples; Quantitate miRNA by qRT-PCR. - [Read Detect and Quantitate MicroRNA in Laser Capture Microdissection Samples]

Category: RNAi Technology Protocols

Information on how detect and quantitate MicroRNA in laser capture microdissection samples. Includes: Obtain Laser Capture Microdissected Samples; Isolate miRNA from LCM Samples; Quantitate miRNA by qRT-PCR; Detection of miRNA in Microdissected Tissue from Mouse Brain by qRT-PCR; Differential Expression of MicroRNA in Whole Brain Tissue Compared to a More Homogeneous Population of Cells. - [Read Detect and Quantitate MicroRNA in Laser Capture Microdissection Samples]

Category: RNA Protocols: RT-PCR and cDNA synthesis

Gene-specific RT-PCR. Used for microdissected RNA, but may apply to other conditions. Molecular Profiling Initiative, NCI. - [Read Gene-specific RT-PCR]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Useful techniques to circumvent disruption of tissue structure in the analysis of gene expression are LCM and LDM. While they require specialized microscopes and systems, they are similar in that freshly-cut frozen tissue sections can be microdissected using either a general histological stain (like H&E) or by staining with fluorescently conjugated antibodies. The LCM system by Arcturus involves... - [Read Immunofluorescent Staining for the Laser Microdissection of Individual Cells Protocol]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Information on processing of microdissected tissue for molecular analysis. Includes: More than 10,000 Cells; Less than 10,000 Cells; Formalin-fixed; Paraffin-embedded Tissue; Other Fixatives; Timing. - [Read Processing of Microdissected Tissue for Molecular Analysis]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

The methodology and buffers utilized for processing microdissected samples for protein-based studies is variable depending on the downstream molecular analysis method. - [Read Processing of Microdissected Tissue for Molecular Analysis Proteomics-based Studies]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

RNA-based studies of processing of microdissected tissue for molecular analysis. Includes: Total RNA Isolation; Optional DNAse Treatment. - [Read Processing of Microdissected Tissue for Molecular Analysis RNA-based Studies]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocols for LCM preparation and analysis. Includes protocols: Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections; Preparation and LCM of Paraffin Embedded Tissue Sections; Standard Protocols for Microdissected Tissue Analysis. - [Read Protocols for LCM Preparation and Analysis]

Category: Primary Cell Isolation and Culture Protocols

The protocol is one example of differential gene expression analysis of cells obtained from microdissected tissue. Includes: Microdissection and RNA Isolation; Reverse Transcription; PCR; P.A.G.E.; Sequencing of Differentially Expressed Bands. - [Read Targeted Differential Display Protocol]