microdissection Protocols

Category: RNA Protocols: RT-PCR and cDNA synthesis

Method of synthesizing cDNA from Frozen Tissue Sections Protocol. Includes LCM - Laser Capture Microdissection Protocols. NIH - [Read cDNA from Frozen Tissue Sections and LCM]

Category: PCR Protocols: Real-Time PCR Protocols

Protocol describes the real-time PCR using RNA purified from laser-captured tissues Laser Capture Microdissection (LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cells. - [Read cDNA Synthesis and Real-Time PCR Using RNA from Laser-Captured Cells 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: Histology Protocols: Tissue Sample Sectioning

PREPARATION OF PLANT TISSUE FOR LASER MICRODISSECTION. Schnable Labs, Plant Genomics. - [Read FROZEN SECTIONING PROTOCOL Plant Tissue PDF]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Human embryonic stem cells (hESCs) are pluripotent cells capable of differentiation to representatives of all three germ layers. Includes: Isolation of Primary Mouse Embryo Fibroblasts; Thawing and preparing p1 MEF feeder plates; Preparation of MEF- Conditioned Medium (MEF-CM; Microdissection Passaging of hESCs; Bulk passaging of hESC; Cryopreservation of hESCs; Thawing of hESCs; Karyotyping. - [Read Human Embryonic Stem Cell Protocols]

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

After fixation, frozen sections are immunostained under RNase-free conditions using a rapid three-step streptavidin-biotin technique followed by dehydration. The immunostained sections are ready for LCM. Includes: Development of Immuno-LCM. - [Read Immuno-Laser Capture Microdissection Protocol]

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

Protocol for Immunoblot. Includes: Staining and Laser Capture Microdissection; Protein Separation by Polyacrylamide Gel Electrophoresis; Electrophoretic Transfer To a Membrane (Nylon, PVDF or Nitrocellulose); Primary and Secondary Antibody Incubations; Visualization. - [Read Immunoblot Protocol]

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

LCM utilizes an infrared laser integrated into a standard microscope. A transparent cap is attached to a thermoplastic transparent membrane which lies directly on the surface of a routinely prepared tissue section on a glass slide. The investigator examines the tissue section microscopically and activates the laser when the desired cells underlie the target. This in turn activates the membrane with subsequent binding and procurement of the cells of interest. - [Read Laser Capture Microdissection (LCM)]

Category: Histology Protocols: Laser Capture and Microdissection

Laser Capture Microdissection of Living in vitro Cells. This PDF describes a precise, rapid and convenient Laser Capture Microdissection (LCM) method for the positive selection of living adherent cells and the successful subsequent re-cultivation of homogenous sub-populations. Arcturus. - [Read Laser Capture Microdissection of Living in vitro Cells PDF]

Category: Histology Protocols: Laser Capture and Microdissection

Preparation of Frozen Tissue for LCM, Preparing Sections from Frozen Tissue for Laser Microdissection, Preparing Sections from Fixed, Paraffin-embedded Tissue for Laser Microdissection Protocols and methods. Laser Microdissection Laboratory. UAB. - [Read Laser Capture Microdissection Protocols]

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

LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, etc. Protocol provides a thorough description of LCM techniques, with an emphasis on tips and troubleshooting advice derived from LCM users. The total time required to carry out this protocol is typically 1–1.5 h. - [Read Laser-capture Microdissection Protocol]

Category: Primary Cell Isolation and Culture Protocols

There are several manual methods that can be used to perform tissue microdissection. Techniques using hand-held tools as well as mechanical micromanipulator-based approaches have been described. However, speed and precision are the most important parameters and any method that achieves these is adequate. Investigators should also expect to invest time initially by practicing on 10 to 20 cases to begin to feel comfortable with the technique. - [Read Manual Microdissection]

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

Human tissues are comprised of multiple interacting cell populations in a complex three dimensional arrangement with each cellular phenotype determined by a unique profile of mRNA and protein expression. Before microdissection techniques were developed, the only analysis tools for phenotypic studies were primarily immunohistochemistry and in-situ hybridization. While useful, these tools are limited to single gene analysis and, in general, do not allow qualitative studies. - [Read Microdissection Overview]

Category: Primary Cell Isolation and Culture Protocols

Overview of microdissection. Includes: The Goals of Microdissection; The Development of Microdissection. - [Read Microdissection Overview]

Category: Histology Protocols: Tissue Sample Sectioning

FIXATION, DEHYDRATION, PARAPLAST INFILTRATION AND SECTIONING PROTOCOL WITH TISSUE COLLECTION PROCEDURE USING THE PALM LASER MICRODISSECTION SYSTEM. Schnable Labs, Plant Genomics. - [Read Paraffin Sectioning for LCM Plant Tissue]

Category: Histology Protocols: Laser Capture and Microdissection

SINGLE CELL CAPTURE AND ANALYSIS. NIH LASER CAPTURE MICRODISSECTION PROTOCOLS. Protocol for Laser Capture Microdissection. - [Read SINGLE CELL CAPTURE AND ANALYSIS]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols: Tissue Preparation for LCM Protocols

Protocol for slide preparation for laser capture microdissection (LCM) for subsequent DNA, RNA, and protein analysis. - [Read Slide Preparation for Laser Capture Microdissection (LCM) Protocol]

Category: Primary Cell Isolation and Culture Protocols

Slide Preparation for Manual Microdissection for Subsequent DNA, RNA, and Protein Analysis. Manual microdissection and subsequent molecular analysis can be carried out on slides stained using standard hematoxylin and eosin methods. However, if cell types that are (or are not) expressing a specific protein are required for a study, then more advanced slide preparation methods such as Immuno-LCM may be utilized. - [Read Slide Preparation for Manual Microdissection Protocol]

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]