Histology / Immunohistochemistry techniques protocols and methods

Technique / Histology / Immunohistochemistry techniques

Histology-related protocols new recommended protocol
An extremly useful immunohistochemistry staining protocols by The Diabetes and Endocrinology Research Center (DERC) at the University of Colorado Health Sciences Center in Denver, Colorado. (27 pages PDF) ...

Immunohistochemistry with Whole Mouse Embryos new protocol
Protocol on immunohistochemistry with whole mouse embryos from Sato's lab at University Texas Southwestern medical center at Dallas. The PDF version of the protocol can be downloaded from the above page. ...

Immunostaining with Paraffin Sections new protocol
From Sato's lab at University of Texas Southwestern medical center. Staining procedure for paraffin sections with antibodies to laminin, fibronectin, collagen type IV. PDF version of the protocol is available on the web page. ...

Immunohistochemistry Staining, Frozen Tissues (Acetone-Fixed) protocol
This protocol is used for for either Purified or Biotinylated primary antibodies for immunohistochemical staining of mouse frozen tissue sections. (ebioscience inc.) ...

DETECTION OF b-GALACTOSIDASE AND ALKALINE PHOSPHATASE ACTIVITIES IN TISSUE protocol
By Constance Cepko et al at Cepko lab, Harvard medical school. ...

Preparation Of Paraffin-Embedded Sections For Immunohistochemistry protocol
Protocol from Chemicon. Tissue sections (5-7 ¦Ìm thick) are cut from paraffin-embedded blocks on a microtome and mounted from warm water (40¡ãC) onto adhesive microscope slides. Sections are allowed to dry overnight at room temperature or 40¡ãC ... ... ...

Double Immunohistochemistry protocol
Procedure: First stain HRP Developing solution Second stain AP Developing solution Protocol by Giorgio Cattoretti, Columbia University. ...

Making Fluorescence Mounting Medium (Antifade) new protocol
Recipe and protocol for making Fluorescence Mounting Medium (Antifade). From Spector's lab at Cold Spring Harbor Lab. ...

Acid Cleaning Coverslips Procedure protocol
Short description on how to clean coverslips for immunofluorescent staining. Spector lab at Cold Spring Harbor Lab. ...

Triple Labeling Immunofluorescence Protocols for Tissue Sections and Cell Cultures new protocol
Triple Labeling Immunofluorescence Protocols for Tissue Sections and Cell Cultures. By Ian Jones at University of Bath, UK. ...

Tissue immunolabelling protocol: ABC/DAB protocol
Tissue immunolabelling protocol using ABC/DAB reaction. By Ian Jones at University of Bath, UK. ...

Immunocytochemistry on free-floating sections protocol
Immunocytochemistry on free-floating sections protocol by Ann Marie Johnson, Fisher Scientific. ...

Staining Methods for cell death protocol
Staining method for cell death: necrosis and apoptosis based on morphology after staining. (Finkbeiner Lab, UCSF) ...

GENERAL IMMUNOFLUORESCENCE PROTOCOL protocol
A pdf general protocol on immunofluorescence staining including antigen retrieval methods. Protocol from Rosen lab, Baylor college of medicine. ...

Contrasting and Drying of Cryosections protocol
This step is essential for fine structure preservation as well as for producing enough contrast to visualize subcellular detail in the electron microscope. (Eye Hearing Institute) ...

Enzymatic Protocol 1.0 new protocol
Protocol for enzymatic stain frozen cells and tissues, as well as paraffin-embedded tissues. (R&D Systems) Contents: Overview Materials Sample Preparation Tissue Fixation and Mounting - Cryostat Sections Tissue Fixation and Mounting - Para ...

Double immunofluorescence staining protocol new protocol
A brief protocol introducing double staining fluorescent simultaneous protocol from ABCam inc. A. Cell lines, cytology smears, cytospin preparations. B. Frozen (cryostat) sections. C. Paraffin-embedded sections. ...

Immunostaining Tutorial new recommended review
A nice pdf tutorial with detailed instruction and images on general immunostaining procedure written by Sandy Grimm, PhD from Rosen lab at Baylor college of medicine. This immunostaining protocol is for paraffin embedded tissue sections. ...

Basic Immunohistochemistry Protocol hot recommended site
Placed on WWW by: Michael Serfas General Note: Protocols for immunohistochemistry vary widely, due to the differences between antigens and their recognition by antibody. Some epitopes are destroyed by the high temperatures and organic solvents used in ...

IHC world immunohistochemistry protocol database site
Protocols and recipes about antibody staining, antigen retrieval, blocking solutions, antibody dilution buffers, washing buffers, chromagen substrate solutions, counterstain solutions, special stains, TUNEL staining, general IHC protocols, general histology an ...

Immunostaining Protocol with Video Demonstration new recommended troubleshooting
A step by step immunostaining protocol with video demonstration on how to perform immunocytochemistry fluorescent staining on cell cultures on coverslips. You are recommended to first download their detailed ...

Troubleshooting on immunohistology methods new recommended troubleshooting
Troubleshooting guide on weak staining, overstaining or high background in immunohiostology staining (IHCWorld.com) ...

Immunohistochemistry Troubleshooting Guide new recommended troubleshooting
Troubleshoot problems : Lack of Staining, overstaining and high background. (from R&D systems) ...

Technical notes on confocal and multi-photon imaging sample processing new recommended troubleshooting
Some technical suggestions for the novice or not so novice user regarding a variety of issues related to confocal and multi-photon imaging. (Confocal Microscopy Core Facility at Brigham and Women's Hospital, Havard University) ...

Problem with nonspecific antibody binding troubleshooting
I am currently developing a protocol for immunofluorescent detection of the stress proteins hsp70 and cpn60 on cryosectioned Mytilus mantle tissue. The problem I am having is nonspecific binding of the secondary antibody (conjugated to fluorescein), l ...

Immunocytochemistry double staining with rabbit antibody troubleshooting
I wish to stain fixed cells for two different poteins. I have selective antibodies to each and one which recognises both. Unfortunately, they ae all raised in rabbit. Is there a simple way of labelling the antibodies directly, so that I could double-s ...

Immunofluorescent staining autofluoresence problem troubleshooting
Anyone know how to reduce/abolish autofluorescence on formain fixed, paraffin embedded sections of human endometrium? (Obviously dewaxed before use, but which give a high fluorescent background never having seen any antibody). ...

Red fluorescent DNA staining dye troubleshooting
Currently I am examining living cells with Green Fluorescent Protein and I want to combine this with a red fluorescent dye to visualize DNA in vital cells using confocal laser scanning microscopy. I cannot use Hoechst or other dyes with UV excitation, ...

Fluorescence Microscopy Filter Sets troubleshooting
I understand that GFP has become popular of late. However, I'd like to know how a researcher understands what filter sets and flourochromes are required to view a speciman optimally. If I have a filter set at my disposal, how can I find out specificall ...

Immunocytochemistry: Common problems and some answers new troubleshooting
Results obtained from immunocytochemical labeling can often differ from what was expected. Sometimes this may result in the formulation of new theories but often good results are ignored because of faulty interpretations. (House Ear Institute) ...

Immunohistochemistry Query DB new database
Developed by Dennis M. Frisman. The database allows histologist 1) list the antibodies that can differentiate between tumors entered by the user (e.g., lung adenocarcinoma vs. breast carcinoma), 2) rank the antibodies in terms of their ability to differentiat ...

General Immunohistochemistry Protocol
(Sharma Laboratory at University of Chicago.)
A general IHC staining protocol.

Preparation and Staining of Frozen Tissue Sections
(Pharmingen immunohistochemical staining protocols)
I. Preparation of Frozen Sections for Sectioning. II. Sectioning of Frozen Tissue. III. Standard Immonuhistochemical Staining Procedure for Frozen Sections

Antibody Search Database
(Antibody search engine for all commercially available antibodies from over 300 antibody vendors.)

immunohistochemistry and immunocytochemistry methods books
(Biowww Bookshelf)
Technical books on IHC study

Glataraldehyde for cell fixation
(Forum)
about fixation of the cells with glutaraldehyde

DAPI staining before secondary antibody
(Forum)
DAPI staining problem

in situ hybridization: all turn blue!!
(Forum)
whole mount in situ hybridization problem

double labeling
(Forum)
two different antibodies both polyclonal goat IgG

monoclonal GFAP immunostaining
(Forum)
nonspecific binding of secondary antibody

blocking endogenous biotin
(Forum)
stain PFA-fixed HeLa/Caco cells with a biotinylated primary Ab and detecting with Streptavidin-FITC

Eliminating non-specific staining for ICC immunocytochemistry
(Forum)
problem of non-specific staining

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Last update:

10-May-2008 01:25 am

Related new papers and reviews

Immunohistochemistry.

This unit describes several methods for localizing specific antigens in various tissue and cell preparations using immunohistochemistry (IHC). Protocols describe preparation of suitable material for IHC including fresh, unfixed, frozen tissue specimens; unfixed cells, either freshly isolated or from suspension or adherent cultures, or fixed, paraffin-embedded tissue sections. By careful selection of reagents, it is possible to detect two antigens simultaneously. For antigens that are sensitive to fixative, it may be necessary to unmask the antigen by a new technique called "antigen retrieval". If there is cross-reactivity between the secondary antibody and antigens present in the target cells or tissue, the secondary antibody can be preabsorbed. The different IHC protocols are represented schematically and summarized in a table, which also lists advantages and disadvantages of each approach. Causes of background staining and ways to eliminate it are also discussed.

Direct immunofluorescence and immunohistochemistry in diagnostics of glomerulonephritis.

The needle biopsies from 60 transplanted and native kidneys have been processed and a prospective analysis of pattern, intensity and distribution of immunoglobulin deposits (IgA, IgG and IgM) and complement components (C3c and C1q) identified in these lesions has been carried out by immunohistochemistry with three step immunoperoxidase, in the period from 2000 to 2004. Those deposits were previously detected and analyzed by immunofluorescence. The samples consisted of 30 renal biopsies, previously diagnosed with glomerulonephritis and positive immunofluorescence and 30 renal biopsies without morphologic changes and deposits on immunofluorescence. 78,7% of the analyzed samples showed the identical results of the deposits of immunoglobulin and components of the complement with both, immunohistochemistry and immunofluorescence method. Sensitivity of the immunohistochemistry method with three step immunoperoxidase for all analyzed immunoglobulin and complement components is high (0,93), while specificity for the same method is 0,79. Standardized method of the three step immunoperoxidase on the paraffin embedded, formalin fixed needle renal biopsies could successfully replace the immunofluorescence method in diagnostic of GN, with the emphasis on a follow up and control of each single step in the procedure of the method.