sections Protocols

Category: Immunohistochemistry Protocols: Antigen Retrieval Protocols

Protocol used to for immunohistochemistry on paraffin-embedded sections. Based on use of microwave energy to effect antigen retrieval. The immunohistochemistry procedure, is for use of Biomeda's HistoScan kit based on a streptavidin-peroxidase/biotinylated second antibody detection system with 3-amino, 9-ethylcarbazole (AEC) as chromogen. Undoubtedly, other kits or home-made reagents will also work . - [Read Antigen Retrieval for Immunohistochemistry with Paraffin-Embedded Tissues Protocol]

Category: Radioactivity: Autoradiography

Antonio Simeone. MRC Centre for Developmental Neurobiology, New Hunt’s Houre. - [Read Autoradiography for mRNA detection in. mouse embryo tissue sections. PDF]

Category: Mouse Protocols

Protocol describes the strongest way for lineage tracing on sections by using biotin dextran amine. - [Read BDA Mouse Lineage Tracing on Sections Protocol]

Category: Neuroscience Protocols: Neuronal Stains Protocols: Stains for Senile Plaques Protocols

A silver stain to demonstrate neurofibrillary tangles, nerve fibers and senile plaques in Alzheimer's disease. The nerve fibers are sensitized with a silver solution.  The sections are treated with ammoniacal silver, and then reduced to a visible metallic silver. - [Read Bielschowsky Technique for Senile Plaques Protocol]

Category: Antibody Protocols: Antibody Labeling Protocols

Once tissues are fixed and permeabilized, the antibodies are added. These antibodies can be labeled directly or detected by a labeled secondary reagent. For indirect detection, any reagent that binds specifically to the primary antibody can be "tagged" and used to locate the antibody. The possible reagents include anti-immunoglobulin antibodies, protein A or G, or, if the first antibody is labeled with biotin, streptavidin. They can be labeled with enzymes or gold. - [Read Binding Antibodies to Tissue Sections Protocol]

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: Signalling Signal Transduction Protocols: Chemokine Protocols

Western Blotting chemokine receptors, Immunohistochemistry on Paraffin sections for chemokines. - [Read Chemokine and Interleukin Antibody Protocols]

Category: Neuroscience Protocols: Neuronal Stains Protocols: Stains for Senile Plaques Protocols

Protocol to demonstrate amyloid deposits in tissue sections. When stained with the Congo Red Stain the amyloid, with the aide of polarizing lenses, will birefringe an apple green color.  Under the microscope. - [Read Congo Red Putcher's Modification Amyloid Protocol]

Category: Cell Culture Protocols: Cell Staining Protocols

Counterstains are used to help differentiate the various cell types of subcellular structures seen in cell staining. They are essential for tissue sections, allowing the identification of the cell types, but also may be helpful in other staining reactions. - [Read Counterstains Protocol]

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for detection of HPV 11 DNA in paraffin-embedded laryngeal tissue with a DIG-labeled DNA probe. Includes: Probe preparation; Pretreatment of slides; Preparation of tissue sections; Hybridization; Washes; Detection of hybrids. - [Read Detection of HPV 11 DNA in Paraffin-Embedded Laryngeal Tissue with a DIG-Labeled DNA Probe Protocol]

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for detection of mRNA in tissue sections using DIG-labeled RNA and oligonucleotide probes. - [Read Detection of mRNA in Tissue Sections Using DIG-labeled RNA and Oligonucleotide Probes Protocol]

Category: Protein Protocols: Protein Extraction From Tissue

Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tissue sections. Ikedaa et al, 1998 JHC. - [Read Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tis]

Category: Lipid Protocols

Protocol for extraction method for formalin pigments from tissue sections. - [Read Extraction Method for Formalin Pigments from Tissue Sections Protocol]

Category: Histology Protocols: Histological Fixation

FORMALDEHYDE-BASED FIXATIVES, PARAFFIN-EMBEDDED SECTIONS, BOUIN’S SOLUTION, MERCURIC CHLORIDE-BASED FIXATIVES. FARMILO et al. - [Read Fixation.]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular.... - [Read Fluorescent In Situ Transcription in Cells and Tissues Protocol]

Category: Drosophila Protocols: Drosophila Dissection Protocols

Protocol for fly head sections using fly collars. - [Read Fly Head Sections Using Fly Collars Protocol]

Category: Gene Delivery Protocols: Biolistics Protocols

The technique has many advantages—plasmids may be used for delivery, DNA theoretically can be delivered to any cell type, and genes may be delivered to cells in vitro, ex vivo, or in vivo. DNA-coated gold particles are distributed evenly along the length of the tubing, which is subsequently cut into short sections of cartridges to be used in a gene gun. The Helios Gene Gun uses a pulse of helium to launch the DNA-coated particles, spreading them onto the target cells. - [Read Gene Delivery to Skin Using Biolistics Protocol]

Category: Molecular Imaging: Immunofluorescence Microscopy

General Immunofluorescence Protocol. Rosen Lab. Deparaffinize and rehydrate sections, Antigen retrieval methods, Proteinase K Antigen Retrieval, Urea Antigen Retrieval, Blocking buffers, - [Read General Immunofluorescence Protocol]

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Protocols for cell fixation and tissue fixation. HISTOLOGY-RELATED PROTOCOLS Fixation and Frozen Block techniques. Embedding Tissue Into Frozen Blocks. Fixing Cells with Fresh Paraformaldehyde. Preparation of Frozen Sections. Immunohistochemistry. Paraffin sections and Frozen sections. DERC Univ. Colorado PDF. - [Read HISTOLOGY-RELATED PROTOCOLS Fixation and Frozen Block PDF]

Category: Immunohistochemistry Protocols: Fixation Protocols

Information on histotechniques. Includes: Fixation - types of fixatives; factors affecting fixation; General usage of fixatives; Tissue Processing; Sectioning; Frozen Sections; Staining; H and E staining; Coverslipping; Decalcification; Artefacts in Histologic Sections; Problems in Tissue Processing. - [Read Histotechniques Fixation]

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: Immunohistochemistry Protocols: Antibody Staining Protocols

There are several strategies to visualize the antibody. For transmitted light microscopy, color development substrates for enzymes are often used. The antibody can be directly labeled with the enzyme. However, such a covalent link between an antibody and an enzyme might result in a loss of both enzyme and antibody activity. For these reasons several multistep staining procedures have been developed, where intermediate link antibodies are used. In this protocol use the Vectastain ABC-kit. - [Read Immunocytochemistry in Free-Floating Sections Protocol]

Category: Molecular Imaging: Immunofluorescence Microscopy

B or T cells in suspension, adherent cells on chambered coverglass or chamberslides, cryostat sections of unfixed, OCT embedded tissue. Susan Anderson. - [Read Immunofluorescence / confocal microscopy]

Category: Immunology: Immunofluorescence Protocols

Protocol for immunofluorescence staining. Includes: Cell Lines; Frozen Sections; Paraffin Sections; Pretreatments of Tissue Sections. - [Read Immunofluorescence Staining 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]