State of the art in antigen retrieval for immunohistochemistry.

The masking effects of antigens by chemical fixation, processing, embedding media interactions, represent a serious problem for immunohistochemical purposes. Fortunately, different approaches in antigen retrieval exist. These techniques are relatively recent and continuously expanding. This review focuses on the present state of the art in antigen retrieval methods for immunohistochemistry in light and electron microscopy. Moreover, a brief discussion on the chemical aspects of fixation, mechanism of retrieval, as well as its efficacy, is given.

[1]  K. Imahori,et al.  Immunocytochemistry of formalin-fixed human brain tissues: microwave irradiation of free-floating sections. , 1998, Brain research. Brain research protocols.

[2]  M. Wick,et al.  Fixation and Epitope Retrieval in Diagnostic Immunohistochemistry: A Concise Review with Practical Considerations , 2000, Applied immunohistochemistry & molecular morphology : AIMM.

[3]  M. Boon,et al.  Microwaves for immunohistochemistry. , 1994, Micron.

[4]  M. Waxham,et al.  Postembedding immunocytochemical localization of paramyxovirus antigens by light and electron microscopy. , 1982, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[5]  S. Park,et al.  Evaluation of Antigen Retrieval Buffer Systems , 2003, Journal of Molecular Histology.

[6]  R. Keijzer,et al.  Routine Formaldehyde Fixation Irreversibly Reduces Immunoreactivity of Bcl-2 in the Nuclear Compartment of Breast Cancer Cells, but not in the Cytoplasm , 2001, Applied immunohistochemistry & molecular morphology : AIMM.

[7]  Y. Okada,et al.  Application of Heat-induced Antigen Retrieval to Aldehyde-fixed Fresh Frozen Sections , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[8]  J. Gu,et al.  Antigen Retrieval Techniques: Immunohistochemistry and Molecular Morphology , 2000 .

[9]  B. Lane,et al.  DIFFERENTIAL STAINING OF ULTRATHN SECTIONS OF EPON-EMBEDDED TISSUES FOR LIGHT MICROSCOPY , 1965, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[10]  Stephen W Carmichael,et al.  Localization of Caveolin 1 in Aortic Valve Endothelial Cells Using Antigen Retrieval , 2002, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[11]  D. Hecke Routine Immunohistochemical Staining Today: Choices to Make, Challenges to Take , 2002 .

[12]  C. Sarraf Immunolabeling for electron microscopy. , 2000, Methods in molecular medicine.

[13]  A. Dodson Modern methods for diagnostic immunocytochemistry , 2002 .

[14]  K. Gatter,et al.  p53 expression in normal paraffin‐embedded tissue using different antibodies and antigen retrieval buffer systems , 2003, Histopathology.

[15]  A. Gown,et al.  Unmasking the mysteries of antigen or epitope retrieval and formalin fixation. , 2004, American journal of clinical pathology.

[16]  L. Larsson,et al.  Improved immunocytochemical detection of daunomycin , 2007, Histochemistry and Cell Biology.

[17]  Chiu Ky Use of microwaves for rapid immunoperoxidase staining of paraffin sections. , 1987 .

[18]  H. Fraenkel-conrat,et al.  The reaction of formaldehyde with proteins; demonstration of intermolecular cross-linking by means of osmotic pressure measurements. , 1949, The Journal of biological chemistry.

[19]  A. Leong,et al.  Superheating Antigen Retrieval , 2002, Applied immunohistochemistry & molecular morphology : AIMM.

[20]  S. Huang Immunohistochemical demonstration of hepatitis B core and surface antigens in paraffin sections. , 1975, Laboratory investigation; a journal of technical methods and pathology.

[21]  R. King,et al.  Ultrasound-accelerated Tissue Fixation/Processing Achieves Superior Morphology and Macromolecule Integrity with Storage Stability , 2006, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[22]  M. Namiki,et al.  Application of immunocryoultramicrotomy to free cells: Ultrastructural localization of myeloperoxidase in neutrophils of human peripheral blood , 1992, American journal of hematology.

[23]  R. Cote,et al.  Antigen Retrieval Technique: A Novel Approach to Immunohistochemistry on Routinely Processed Tissue Sections , 1996 .

[24]  R. Cote,et al.  Calcium-induced Modification of Protein Conformation Demonstrated by Immunohistochemistry: What Is the Signal? , 1999, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[25]  Thomas E. Creighton,et al.  Protein function : a practical approach , 1989 .

[26]  M. Mclaughlin,et al.  Antigen retrieval in cryostat tissue sections and cultured cells by treatment with sodium dodecyl sulfate (SDS) , 1996, Histochemistry and Cell Biology.

[27]  G. C. Moriarty ADENOHYPOPHYSIS: ULTRASTRUCTURAL CYTOCHEMISTRY A REVIEW , 1973, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[28]  H. Fraenkel-conrat,et al.  Reaction of formaldehyde with proteins; cross-linking of amino groups with phenol, imidazole, or indole groups. , 1948, The Journal of biological chemistry.

[29]  J. Povlishock,et al.  The visualization of a new class of traumatically injured axons through the use of a modified method of microwave antigen retrieval , 1999, Acta Neuropathologica.

[30]  C. Taylor Standardization in immunohistochemistry: the role of antigen retrieval in molecular morphology , 2006, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[31]  D. Rampling,et al.  Paraffin wax embedded muscle is suitable for the diagnosis of muscular dystrophy , 2001, Journal of clinical pathology.

[32]  S. Brorson The combination of high-accelerator epoxy resin and antigen retrieval to obtain more intense immunolabeling on epoxy sections than on LR-white sections for large proteins. , 1998, Micron.

[33]  H. Fraenkel-conrat,et al.  The reaction of formaldehyde with proteins; participation of indole groups; gramicidin. , 1947, The Journal of biological chemistry.

[34]  M. Rubin,et al.  Inadequate formalin fixation decreases reliability of p27 immunohistochemical staining: probing optimal fixation time using high-density tissue microarrays. , 2002, Human pathology.

[35]  I. Clemmensen,et al.  The value of hyaluronidase treatment of different tissues before demonstration of fibronectin by the indirect immunoperoxidase technique , 2004, Histochemistry.

[36]  Y. Shibuya,et al.  Comparison of immunostainings for proliferating cell nuclear antigen and Ki-67 in human extraocular lesions , 1997, Graefe's Archive for Clinical and Experimental Ophthalmology.

[37]  N. Singh,et al.  Immunohistochemical analysis of Hodgkin's disease using microwave heating. , 1993, Journal of clinical pathology.

[38]  M. Hauer-Jensen,et al.  Effects of Fixative and Fixation Time for Quantitative Computerized Image Analysis of Immunohistochemical Staining , 1999 .

[39]  W. Hennink,et al.  Identification of Formaldehyde-induced Modifications in Proteins , 2004, Journal of Biological Chemistry.

[40]  W. Grizzle,et al.  Methods of Antigen Recovery Vary in Their Usefulness in Unmasking Specific Antigens in Immunohistochemistry , 2000, Applied immunohistochemistry & molecular morphology : AIMM.

[41]  S. Brorson Improved immunogold labeling of epoxy sections by the use of propylene oxide as additional agent in dehydration, infiltration and embedding. , 1996, Micron.

[42]  D. French,et al.  The Reactions of Formaldehyde with Amino Acids and Proteins , 1945 .

[43]  A. Rhodes Quality assurance in immunohistochemistry. , 2003, The American journal of surgical pathology.

[44]  L. Rangell,et al.  Application of Microwave Technology to the Processing and Immunolabeling of Plastic-embedded and Cryosections , 2000, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[45]  S. Brorson Heat-induced antigen retrieval of epoxy sections for electron microscopy. , 2001, Histology and histopathology.

[46]  K. Schmid,et al.  Possible role of tissue‐bound calcium ions in citrate‐mediated high‐temperature antigen retrieval , 1994, The Journal of pathology.

[47]  S. Cruwys,et al.  An evaluation of the immunohistochemistry benefits of boric acid antigen retrieval on rat decalcified joint tissues. , 2007, Journal of immunological methods.

[48]  M. Mehrabian,et al.  Contact-site cross-linking agents , 1981, Molecular and Cellular Biochemistry.

[49]  R. Warnke,et al.  Suppression of endogenous avidin-binding activity in tissues and its relevance to biotin-avidin detection systems. , 1981, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[50]  H. Uylings,et al.  Antigen retrieval in formaldehyde-fixed human brain tissue. , 1998, Methods.

[51]  B. Rosario,et al.  A SIMPLE METHOD FOR REMOVING THE RESIN FROM EPOXY-EMBEDDED TISSUE , 1961, The Journal of biophysical and biochemical cytology.

[52]  M. Werner,et al.  Antigen retrieval, signal amplification and intensification in immunohistochemistry , 1996, Histochemistry and Cell Biology.

[53]  H. Uylings,et al.  An optimal antigen retrieval method suitable for different antibodies on human brain tissue stored for several years in formaldehyde fixative , 1997, Journal of Neuroscience Methods.

[54]  S. Bogen,et al.  A molecular model of antigen retrieval using a peptide array. , 2006, American journal of clinical pathology.

[55]  R. Cote,et al.  Antigen retrieval for immunohistochemistry status and need for greater standardization , 1996 .

[56]  Y. Fukuda,et al.  Optimizing collagen antigen unmasking in paraffin-embedded tissues , 2004, The Histochemical Journal.

[57]  M. C. Pangborn,et al.  THE REACTION OF FORMALDEHYDE WITH AMINO ACIDS , 1936 .

[58]  R. Cote,et al.  Use of pH 9.5 Tris-HCl buffer containing 5% urea for antigen retrieval immunohistochemistry. , 1996, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[59]  J. Nesland,et al.  Ultrastructural and electron immunohistochemical features of medullary thyroid carcinoma , 2004, Virchows Archiv A.

[60]  J. Baselga,et al.  Pharmacodynamic studies of gefitinib in tumor biopsy specimens from patients with advanced gastric carcinoma. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[61]  P. V. von Hippel,et al.  Formaldehyde as a probe of DNA structure. r. Mechanism of the initial reaction of Formaldehyde with DNA. , 1977, Biochemistry.

[62]  J. M. Robinson,et al.  Antigen retrieval in cells and tissues: enhancement with sodium dodecyl sulfate , 2001, Histochemistry and Cell Biology.

[63]  R. Horobin Staining plastic sections , 1983 .

[64]  C. Taylor,et al.  Paraffin section immunocytochemistry for estrogen receptor: The time has come , 1996, Cancer.

[65]  C. Rodning,et al.  Technical parameters of immunostaining of osmicated tissue in epoxy sections. , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[66]  A. Leong,et al.  Microwave procedures for electron microscopy and resin-embedded sections. , 1998, Micron.

[67]  Johannes Gerdes,et al.  Monoclonal antibodies against recombinant parts of the Ki‐67 antigen (MIB 1 and MIB 3) detect proliferating cells in microwave‐processed formalin‐fixed paraffin sections , 1992, The Journal of pathology.

[68]  S. Brorson A method for measurements of the efficiency of immunogold labelling of epoxy-embedded proteins subjected to different retrieval techniques. , 2004, Micron.

[69]  P. Ruck,et al.  A comparison of methods for heat-mediated antigen retrieval for immunoelectron microscopy: demonstration of cytokeratin No. 18 in normal and neoplastic hepatocytes. , 1996, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[70]  T. Wight,et al.  Colloidal gold immunostaining on deplasticized ultra-thin sections. , 1988, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[71]  G. Houen,et al.  Interference of sugars with the binding of biotin to streptavidin and avidin. , 1997, Journal of immunological methods.

[72]  H. Furuoka,et al.  Effective antigen-retrieval method for immunohistochemical detection of abnormal isoform of prion proteins in animals , 2005, Acta Neuropathologica.

[73]  G. H. Nguyen,et al.  Increased level of immunogold labeling of epoxy sections by rising the temperature significantly beyond 100 degrees C in the antigen retrieval medium. , 2001, Micron.

[74]  L. Hafer,et al.  Antibodies immunoreactive with formalin-fixed tissue antigens recognize linear protein epitopes. , 2006, American journal of clinical pathology.

[75]  A. Johnson,et al.  Ultrastructural immunogold labeling of glial filaments in osmicated and unosmicated epoxy-embedded tissue. , 1990, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[76]  Johannes Gerdes,et al.  Antigen unmasking on formalin‐fixed, paraffin‐embedded tissue sections , 1993, The Journal of pathology.

[77]  M. Key,et al.  Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections. , 1991, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[78]  D. Vandré,et al.  Efficient Immunocytochemical Labeling of Leukocyte Microtubules with FluoroNanogold: An Important Tool for Correlative Microscopy , 1997, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[79]  S. Vidal,et al.  An easy method for the removal of Epon resin from semi-thin sections. Application of the avidin-biotin technique , 1995, The Histochemical Journal.

[80]  T. Kosaka,et al.  Use of high concentrations of glutaraldehyde for immunocytochemistry of transmitter-synthesizing enzymes in the central nervous system , 1986, Neuroscience.

[81]  Y. Okada,et al.  Mechanisms of heat-induced antigen retrieval: analyses in vitro employing SDS-PAGE and immunohistochemistry. , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[82]  D. Hopwood,et al.  Differentiating the effects of microwave and heat on tissue proteins and their crosslinking by formaldehyde , 1988, The Histochemical Journal.

[83]  M. Silver,et al.  Immunogold detection of chromogranin A in the neuroendocrine tumor. , 1993, Ultrastructural pathology.

[84]  M. Ding,et al.  The microtubule cytoskeleton in human phagocytic leukocytes is a highly dynamic structure. , 1995, European journal of cell biology.

[85]  P. di Sant'Agnese,et al.  Androgen receptor immunostaining and its tissue distribution in formalin-fixed, paraffin-embedded sections after microwave treatment. , 1994, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[86]  A. R. Hansen,et al.  A comparative study of the immunogold labeling on H(2)O(2)-treated and heated epoxy sections. , 2001, Micron.

[87]  S. Yamashita,et al.  Immunolocalization of Aldolase A Subunit Using Monoclonal Antibody in Rabbit Tissues , 1997 .

[88]  K. Dill,et al.  Denatured states of proteins. , 1991, Annual review of biochemistry.

[89]  C. Dreyer,et al.  Urea reactivates antigens in paraffin sections for immunofluorescent staining. , 1982, Stain technology.

[90]  S. Pfeiffer Use of hydrogen peroxide to accelerate staining of ultrathin Spurr sections. , 1982, Stain technology.

[91]  M. T. Silva Electron microscopic study on the effect of the oxidation of ultrathin sections of Bacillus cereus and Bacillus megaterium. , 1967, Journal of ultrastructure research.

[92]  A. Bateman,et al.  immunocytochemistry: a study of 80 antibodies , 2022 .

[93]  J. Southgate,et al.  Expression of an Antigen Associated with Basal Bodies of Human Ciliated Epithelial Cells , 2004, The Histochemical Journal.

[94]  B. Soltys,et al.  Interrelationships of endoplasmic reticulum, mitochondria, intermediate filaments, and microtubules--a quadruple fluorescence labeling study. , 1992, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[95]  R. Cote,et al.  A Modified Reduced-Temperature Antigen Retrieval Protocol Effective for Use With a Polyclonal Antibody to Cyclooxygenase-2 (PG 27) , 2002, Applied immunohistochemistry & molecular morphology : AIMM.

[96]  D. J. Long,et al.  Microwave oven-based technique for immunofluorescent staining of paraffin-embedded tissues , 2008, Journal of Molecular Histology.

[97]  T. Nunoya,et al.  Variations in effectiveness of antigen retrieval pretreatments for diagnostic immunohistochemistry. , 1998, Research in veterinary science.

[98]  J. Pollock,et al.  The detection of CD2+, CD4+, CD8+, and WC1+ T lymphocytes, B cells and macrophages in fixed and paraffin embedded bovine tissue using a range of antigen recovery and signal amplification techniques. , 1999, Veterinary immunology and immunopathology.

[99]  R. Ravid,et al.  Activation of the human supraptic and paraventricular nucleus neurons with aging and in Alzheimer's disease as judged from increasing size of the Golgi apparatus , 1993, Brain Research.

[100]  Young Kee Shin,et al.  The Enhanced Reactivity of Endogenous Biotin-like Molecules by Antigen Retrieval Procedures and Signal Amplification with Tyramine , 2002, The Histochemical Journal.

[101]  S. Brorson,et al.  Mechanism for antigen detection on deplasticized epoxy sections. , 1995, Micron.

[102]  H. Sugimura,et al.  Alteration of immunoreactivity by hydrated autoclaving, microwave treatment, and simple heating of paraffin‐embedded tissue sections , 1994, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[103]  M. Haritani,et al.  Hydrated autoclave pretreatment enhancement of prion protein immunoreactivity in formalin-fixed bovine spongiform encephalopathy-affected brain , 2004, Acta Neuropathologica.

[104]  L. Larsson Immunocytochemistry: Theory and Practice , 1988 .

[105]  J. Roth,et al.  Unconventional antigen retrieval for carbohydrate and protein antigens , 1998, Histochemistry and Cell Biology.

[106]  M. Maxwell Two rapid and simple methods used for the removal of resins from 1.0 μm thick epoxy sections , 1978, Journal of microscopy.

[107]  E. Chi‐Ahumada,et al.  Application of the Critical Molar Concentration Concept to Heat-mediated Antigen Retrieval in Immunohistochemistry , 2000, The Histochemical Journal.

[108]  T. Ezaki,et al.  The identification of proliferating cell nuclear antigen (PCNA) on rat tissue cryosections and its application to double immunostaining with other markers. , 1995, Archives of histology and cytology.

[109]  Dr. Gareth Griffiths Fine Structure Immunocytochemistry , 1993, Springer Berlin Heidelberg.

[110]  M. Kook,et al.  Optimal Conditions for the Retrieval of CD4 and CD8 Antigens in Formalin-Fixed, Paraffin-Embedded Tissues , 2003, Journal of Molecular Histology.

[111]  S. Bogen,et al.  A high throughput combinatorial library technique for identifying formalin-sensitive epitopes. , 2006, Journal of immunological methods.

[112]  Clive R Taylor,et al.  Recommendations for Improved Standardization of Immunohistochemistry , 2007, Applied immunohistochemistry & molecular morphology : AIMM.

[113]  T. Boenisch Formalin-Fixed and Heat-Retrieved Tissue Antigens: A Comparison of Their Immunoreactivity in Experimental Antibody Diluents , 2001, Applied immunohistochemistry & molecular morphology : AIMM.

[114]  T. Boenisch Effect of Heat-Induced Antigen Retrieval Following Inconsistent Formalin Fixation , 2005, Applied immunohistochemistry & molecular morphology : AIMM.

[115]  T. Iwadare,et al.  A solution for removal of resin from epoxy sections. , 1990, Stain technology.

[116]  C. Chang,et al.  Immunostaining of cytology smears: A comparative study to identify the most suitable method of smear preparation and fixation with reference to commonly used immunomarkers , 2003, Diagnostic cytopathology.

[117]  J. Hendricks,et al.  Effect of fixation time and microwave oven heating time on retrieval of the Ki-67 antigen from paraffin-embedded tissue. , 1993, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[118]  P. Maxwell,et al.  Audit and internal quality control in immunohistochemistry , 2000, Journal of clinical pathology.

[119]  R. Cote,et al.  Antigen retrieval immunohistochemistry used for routinely processed celloidin-embedded human temporal bone sections: standardization and development. , 1998, Auris, nasus, larynx.

[120]  K. Imahori,et al.  Immunocytochemistry of tau phosphoserine 413 and tau protein kinase I in Alzheimer pathology , 1996, Brain Research.

[121]  C. Foster,et al.  Primer: tissue fixation and preservation for optimal molecular analysis of urologic tissues , 2006, Nature Clinical Practice Urology.

[122]  Y. Okada,et al.  Mechanisms of Heat-induced Antigen Retrieval: Does pH or Ionic Strength of the Solution Play a Role for Refolding Antigens? , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[123]  J. Petrali,et al.  Immunolocalization of MAP-2 in Routinely Formalin-Fixed, Paraffin-Embedded Guinea Pig Brain Sections Using Microwave Irradiation: A Comparison of Different Combinations of Antibody Clones and Antigen Retrieval Buffer Solutions , 2005, Microscopy and Microanalysis.

[124]  Anthony S-Y Leong,et al.  How Does Antigen Retrieval Work? , 2007, Advances in anatomic pathology.

[125]  B. Mepham,et al.  The use of proteolytic enzymes to improve immunoglobulin staining by the PAP technique , 1979, The Histochemical Journal.

[126]  G. Jacobsen,et al.  Microwave processing for immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA) in formalin‐fixed and paraffin‐embedded tissue , 1994, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[127]  S. Brorson Deplasticizing or etching of epoxy sections with different concentrations of sodium ethoxide to enhance the immunogold labeling. , 2001, Micron.

[128]  S. Brorson,et al.  Increased yield of immunogold labeling of epoxy sections by adding para-phenylendiamine in the tissue processing. , 1999, Micron.

[129]  T. Hashimoto,et al.  Immunocytochemical localization of peroxisomal enzymes in human liver biopsies. , 1987, The American journal of pathology.

[130]  H. Ino Antigen Retrieval by Heating En Bloc for Pre-fixed Frozen Material , 2003, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[131]  Brian J Cummings,et al.  Optimization of techniques for the maximal detection and quantification of Alzheimer’s-related neuropathology with digital imaging , 2002, Neurobiology of Aging.

[132]  A. Chott,et al.  Effect of formalin tissue fixation and processing on immunohistochemistry. , 2000, The American journal of surgical pathology.

[133]  I. Baas,et al.  Detection of p53 overexpression in routinely paraffin-embedded tissue of human carcinomas using a novel target unmasking fluid. , 1993, The American journal of pathology.

[134]  E. Kellenberger,et al.  The efficiency of immunolabel on Lowicryl sections compared to theoretical predictions. , 1987, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[135]  M. Engel,et al.  Paediatric rhabdomyosarcoma: MyoD1 demonstration in routinely processed tissue sections using wet heat pretreatment (pressure cooking) for antigen retrieval. , 1997, Journal of clinical pathology.

[136]  G. Bullock,et al.  Techniques in Immunocytochemistry , 1983 .

[137]  R. Curran,et al.  The unmasking of antigens in paraffin sections of tissue by trypsin , 1977, Experientia.

[138]  K. Helander,et al.  Kinetic studies of formaldehyde binding in tissue. , 1994, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[139]  K. Kitahara,et al.  Immunohistochemical detection of retinal cones in monkey retina: light and electron microscopic study. , 2000, Japanese journal of ophthalmology.

[140]  T. Ezaki [Antigen retrieval: its significance and drawbacks in immunohistochemistry]. , 1996, Kaibogaku zasshi. Journal of anatomy.

[141]  S. Vidal,et al.  An easy method for the removal of Epon resin from semi-thin sections. Application of the avidin-biotin technique , 2004, The Histochemical Journal.

[142]  M. Boon,et al.  Notes on the application of microwaves for antigen retrieval in paraffin and plastic tissue sections. , 1993, European journal of morphology.

[143]  S. Prusiner,et al.  Formic acid pretreatment enhances immunostaining of cerebral and systemic amyloids. , 1987, Laboratory investigation; a journal of technical methods and pathology.

[144]  J. Roth Post‐embedding cytochemistry with gold‐labelled reagents: a review , 1986, Journal of microscopy.

[145]  Shuji Yamashita,et al.  Heat-induced antigen retrieval: mechanisms and application to histochemistry. , 2007, Progress in histochemistry and cytochemistry.

[146]  R. Larson,et al.  Effects of Delayed or Prolonged Fixation on Immunohistochemical Detection of Bovine Viral Diarrhea Virus Type I in Skin of Two Persistently Infected Calves , 2005, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[147]  J. Stirling,et al.  Antigen unmasking for immunoelectron microscopy: labeling is improved by treating with sodium ethoxide or sodium metaperiodate, then heating on retrieval medium. , 1995, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[148]  T. Hashimoto,et al.  Light microscopic immunocytochemical demonstration of peroxisomal enzymes in epon sections , 2004, Histochemistry.

[149]  J. Baker Cytological technique : the principles underlying routine methods , 1960 .

[150]  P. Maxwell,et al.  Immunohistochemical staining of plastic embedded bone marrow trephine biopsy specimens after microwave heating. , 1995, Journal of clinical pathology.

[151]  S. Brorson pH-dependent effect of heat-induced antigen retrieval of epoxy section for electron microscopy. , 2002, Micron.

[152]  R. Cote,et al.  Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections. , 1993, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[153]  M. Werner,et al.  Effects of antigen retrieval by microwave heating in formalin-fixed tissue sections on a broad panel of antibodies , 1994, Histochemistry.

[154]  J. Ward,et al.  Immunohistochemical Markers for the Rodent Immune System , 2006, Toxicologic pathology.

[155]  S. Brorson,et al.  Improved technique for immunoelectron microscopy. How to prepare epoxy resin to obtain approximately the same immunogold labeling for epoxy sections as for acrylic sections without any etching. , 1996, Micron.

[156]  A. Reiner,et al.  A simple and sensitive antigen retrieval method for free-floating and slide-mounted tissue sections , 1999, Journal of Neuroscience Methods.

[157]  W. Grizzle,et al.  Advanced Concepts in Fixation: 1. Effects of Fixation on Immunohistochemistry, Reversibility of Fixation and Recovery of Proteins, Nucleic Acids, and other Molecules from Fixed and Processed Tissues. 2. Developmental Methods of Fixation , 2001 .

[158]  F. Reinholt,et al.  Heat-induced retrieval of immunogold labeling for nucleobindin and osteoadherin from Lowicryl sections of bone. , 2006, Micron.

[159]  H. Uylings,et al.  Microwave-stimulated antigen retrieval is pH and temperature dependent. , 1994, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[160]  J. Whitehead,et al.  Prevention of nonspecific binding of avidin. , 1990, Methods in enzymology.

[161]  L. Luciano,et al.  Re-evaluation of Epoxy Resin Sections for Light and Electron Microscopic Immunostaining , 2001, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[162]  L. Kääriäinen,et al.  A method for exposing hidden antigenic sites in paraformaldehyde-fixed cultured cells, applied to initially unreactive antibodies. , 1993, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[163]  P. Roller,et al.  Formaldehyde fixation. , 1985, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[164]  H. Shin,et al.  Methods in pathology. Optimization of proliferating cell nuclear antigen immunohistochemical staining by microwave heating in zinc sulfate solution. , 1994, Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc.

[165]  M. Hayat Microscopy, Immunohistochemistry, and Antigen Retrieval Methods: For Light and Electron Microscopy , 2002 .

[166]  M. Arenas,et al.  Ultrastructural Staining with Sodium Metaperiodate and Sodium Borohydride , 2002, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[167]  S. Yokota,et al.  Immunofluorescence technique for 100-nm-thick semithin sections of Epon-embedded tissues , 2001, Histochemistry and Cell Biology.

[168]  A. Katoh,et al.  Nonspecific Antigen Retrieval Solutions , 1994 .

[169]  O. Ottersen Quantitative electron microscopic immunocytochemistry of neuroactive amino acids , 2004, Anatomy and Embryology.

[170]  A. Pearse Histochemistry: Theoretical and Applied , 1953 .

[171]  R. Brown,et al.  Utility of microwave-citrate antigen retrieval in diagnostic immunohistochemistry. , 1995, Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc.

[172]  K. Kashima,et al.  An Antigen Retrieval Method Using an Alkaline Solution Allows Immunoelectron Microscopic Identification of Secretory Granules in Conventional Epoxy-embedded Tissue Sections , 2003, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[173]  R. Cote,et al.  Antigen Retrieval Immunohistochemistry: Past, Present, and Future , 1997, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[174]  H. Battifora,et al.  The influence of protease digestion and duration of fixation on the immunostaining of keratins. A comparison of formalin and ethanol fixation. , 1986, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[175]  S. Prusiner,et al.  Immuno-gold localization of prion filaments in scrapie-infected hamster brains. , 1987, Laboratory investigation; a journal of technical methods and pathology.

[176]  C. Balaban,et al.  Immunohistochemistry of Lymphocytes and Macrophages in Human Celloidin-Embedded Temporal Bone Sections with Acute Otitis Media , 1997, The Annals of otology, rhinology, and laryngology.

[177]  Huang Sn Immunohistochemical demonstration of hepatitis B core and surface antigens in paraffin sections. , 1975 .

[178]  T. O'Leary,et al.  Effects of formaldehyde fixation on protein secondary structure: a calorimetric and infrared spectroscopic investigation. , 1991, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[179]  A. Douglas-Jones,et al.  The effect of fixation and processing on the sensitivity of oestrogen receptor assay by immunohistochemistry in breast carcinoma. , 2002, Journal of clinical pathology.

[180]  A. Leong,et al.  The Effects of Progressive Formaldehyde Fixation on the Preservation of Tissue Antigens , 1989, Pathology.

[181]  J. Manning,et al.  Effect of Trypsinization on the Immunostaining of Formalin‐Fixed, Paraffin‐Embedded Tissues , 1988, The American journal of surgical pathology.

[182]  E. StrØM,et al.  Immunoelectron microscopy on epoxy sections without deplasticizing to detect glomerular immunoglobulin and complement deposits in renal diseases , 1997, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[183]  C. Hann,et al.  Antigen Retrieval of Basement Membrane Proteins from Archival Eye Tissues , 2001, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[184]  K. Stacey,et al.  THE REACTIONS OF THE “RADIOMIMETIC” ALKYLATING AGENTS WITH MACROMOLECULES IN VITRO , 1958, Annals of the New York Academy of Sciences.

[185]  H. Puchtler,et al.  On the chemistry of formaldehyde fixation and its effects on immunohistochemical reactions , 2004, Histochemistry.

[186]  T. Boenisch Heat-induced Antigen Retrieval: What Are We Retrieving? , 2006, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[187]  M. Shires,et al.  Post-embedding Double-labeling of Antigen-retrieved Ultrathin Sections Using a Silver Enhancement-controlled Sequential Immunogold (SECSI) Technique , 2004, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[188]  M. Bendayan,et al.  Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. , 1983, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[189]  T. Kitamoto,et al.  Immunoreactivity of cerebral amyloidosis is enhanced by protein denaturation treatments , 1991, Acta Neuropathologica.

[190]  M. A. Hayat,et al.  Principles and Techniques of Electron Microscopy , 1975 .

[191]  S. Pileri,et al.  Antigen retrieval techniques in immunohistochemistry: comparison of different methods , 1997, The Journal of pathology.

[192]  J. Hata,et al.  Immunohistochemical Identification of Aggregated Actin Filaments in Formalin‐Fixed, Paraffin‐Embedded Sections: I. A Study of Infantile Digital Fibromatosis by a New Pretreatment , 1992, The American journal of surgical pathology.

[193]  A. Nanci,et al.  Effect of tissue processing on colloidal gold cytochemistry. , 1987, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[194]  J. Hauw,et al.  PrP immunohistochemistry: Different protocols, including a procedure for long formalin fixation, and a proposed schematic classification for deposits in sporadic Creutzfeldt‐Jakob disease , 2000, Microscopy research and technique.

[195]  B. K. Hartman,et al.  Importance of fixation in immunohistochemistry: use of formaldehyde solutions at variable pH for the localization of tyrosine hydroxylase. , 1981, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[196]  R. Allison,et al.  p53, PCNA and Ki-67 expression in oral squamous cell carcinomas: the vagaries of fixation and microwave enhancement of immunocytochemistry. , 2007, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[197]  A. Feller,et al.  Constant detection of surface and cytoplasmic immunoglobulin heavy and light chain expression in formalin‐fixed and paraffin‐embedded material , 1993, The Journal of pathology.

[198]  T J O'Leary,et al.  Standardization in Immunohistochemistry , 2001, Applied immunohistochemistry & molecular morphology : AIMM.

[199]  E. Christensen,et al.  Immunocytochemistry of renal membrane proteins on epoxy sections. , 2007, Kidney international.

[200]  B. Jasani,et al.  ROLE OF CALCIUM CHELATION IN HIGH‐TEMPERATURE ANTIGEN RETRIEVAL AT DIFFERENT pH VALUES , 1997, The Journal of pathology.

[201]  A. Roessner,et al.  An Evaluation of Antigen Retrieval Procedures for Immunoelectron Microscopic Classification of Amyloid Deposits , 1999, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[202]  Y. Sugisaki,et al.  Reversing the Effects of Formalin Fixation with Citraconic Anhydride and Heat: A Universal Antigen Retrieval Method , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[203]  B. Jasani,et al.  The role and mechanism of high-temperature antigen retrieval in diagnostic pathology , 2001 .

[204]  M. Asaka,et al.  Antigen Retrieval Trial for Post-embedding Immunoelectron Microscopy by Heating with Several Unmasking Solutions , 2003, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[205]  S. Huang,et al.  Application of immunofluorescent staining on paraffin sections improved by trypsin digestion. , 1976, Laboratory investigation; a journal of technical methods and pathology.

[206]  R. Merriam The Contribution of Lower Oxides of Osmium to the Density of Biological Specimens in Electron Microscopy , 1958, The Journal of biophysical and biochemical cytology.

[207]  S. Yamashita,et al.  Immunohistochemical study of. GAMMA.-glutamyl transpeptidase with monoclonal antibodies. I Preparation and characteristics of monoclonal antibodies to. GAMMA.-glutamyl transpeptidase , 1986 .

[208]  R. Dapson Fixation for the 1990's: a review of needs and accomplishments. , 1993, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[209]  D. Grube,et al.  Serial semithin sections in immunohistochemistry: techniques and applications. , 1986, Archivum histologicum Japonicum = Nihon soshikigaku kiroku.

[210]  A. Merighi,et al.  Post-embedding immunogold staining , 1993 .

[211]  V. Rait,et al.  Modeling formalin fixation and antigen retrieval with bovine pancreatic ribonuclease A: I—Structural and functional alterations , 2004, Laboratory Investigation.

[212]  A. Tiebosch,et al.  Low-temperature heating overnight in tris-HCl buffer pH 9 is a good alternative for antigen retrieval in formalin-fixed paraffin-embedded tissue , 1998 .

[213]  M. Tabaton,et al.  Formic acid treatment exposes hidden neurofilament and tau epitopes in abnormal cytoskeletal filaments from patients with progressive supranuclear palsy and Alzheimer's disease , 1990, Neuroscience Letters.

[214]  Hector Battifora More on Standardization , 1998 .

[215]  S R Shi,et al.  DEVELOPMENT OF AN OPTIMAL PROTOCOL FOR ANTIGEN RETRIEVAL: A ‘TEST BATTERY’ APPROACH EXEMPLIFIED WITH REFERENCE TO THE STAINING OF RETINOBLASTOMA PROTEIN (pRB) IN FORMALIN‐FIXED PARAFFIN SECTIONS , 1996, The Journal of pathology.

[216]  A. Leong,et al.  Microwave-Stimulated Antigen Retrieval for Immunohistology and Immunoelectron Microscopy of Resin-Embedded Sections , 1998 .

[217]  Clive R Taylor,et al.  Evaluation of the Value of Frozen Tissue Section Used as “Golden Standard” for Immunohistochemistry , 2007, American journal of clinical pathology.

[218]  Mcnicol,et al.  Optimizing immunohistochemistry: antigen retrieval and signal amplification , 1998, Histopathology.

[219]  T. O'Leary,et al.  A molecular mechanism of formalin fixation and antigen retrieval. , 2004, American journal of clinical pathology.

[220]  K. Wong,et al.  Localization of dengue virus in naturally infected human tissues, by immunohistochemistry and in situ hybridization. , 2004, The Journal of infectious diseases.

[221]  M. de Rosa,et al.  Non‐thermal effects of microwaves on proteins: thermophilic enzymes as model system , 1997, FEBS letters.

[222]  S. Yamashita,et al.  IMMUNOHISTOCHEMICAL STUDY OF GAMMA-GLUTAMYL TRANSPEPTIDASE WITH MONOCLONAL ANTIBODIES : II. AN IMMUNOELECTRON MICROSCOPIC STUDY IN RAT KIDNEY , 1989 .

[223]  A. Serizawa,et al.  Heat‐induced antigen retrieval of proliferating cell nuclear antigen and p53 protein in formalin‐fixed, paraffin‐embedded sections , 1994, Pathology international.

[224]  R. Cote,et al.  Comparative study of antigen retrieval heating methods: microwave, microwave and pressure cooker, autoclave, and steamer. , 1996, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[225]  S. DeArmond,et al.  Post-embedding immunoperoxidase staining of glial fibrillary acidic protein for light and electron microscopy , 1981, Journal of Neuroimmunology.

[226]  R. McConnell,et al.  Microwave antigen retrieval for immunocytochemistry on formalin‐fixed, paraffin‐embedded post‐mortem CNS tissue , 1995, The Journal of pathology.

[227]  A. Merighi Post-embedding electron microscopic immunocytochemistry , 1992 .

[228]  J. Said,et al.  Detection of estrogen receptors with monoclonal antibodies in routinely processed formalin-fixed paraffin sections of breast carcinoma. Use of DNase pretreatment to enhance sensitivity of the reaction. , 1987, American journal of clinical pathology.

[229]  F. Mostofi,et al.  Ultrasound-accelerated formalin fixation of tissue improves morphology, antigen and mRNA preservation , 2005, Modern Pathology.

[230]  W. Hennink,et al.  Identification of formaldehyde-induced modifications in proteins: reactions with insulin. , 2006, Bioconjugate chemistry.

[231]  R. E. Cunningham,et al.  ‘Tissue surrogates’ as a model for archival formalin-fixed paraffin-embedded tissues , 2007, Laboratory Investigation.

[232]  L. Kruger,et al.  SELECTIVE STAINING OF NERVOUS TISSUE FOR LIGHT MICROSCOPY FOLLOWING PREPARATION FOR ELECTRON MICROSCOPY , 1968, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[233]  H. Stein,et al.  Use of freeze-dried paraffin-embedded sections for immunohistologic staining with monoclonal antibodies. , 1985, Laboratory investigation; a journal of technical methods and pathology.

[234]  A. Norton,et al.  Brief, high‐temperature heat denaturation (pressure cooking): A simple and effective method of antigen retrieval for routinely processed tissues , 1994, The Journal of pathology.

[235]  K. Tsuneyama,et al.  Enhanced expression of basement‐membrane‐type heparan sulfate proteoglycan in tumor fibro‐myxoid stroma of intrahepatic cholangiocarcinoma , 2001, Pathology international.

[236]  A. Bankfalvi,et al.  Wet autoclave pretreatment for antigen retrieval in diagnostic immunohistochemistry , 1994, The Journal of pathology.

[237]  T Ezaki,et al.  Antigen retrieval on formaldehyde-fixed paraffin sections: its potential drawbacks and optimization for double immunostaining. , 2000, Micron.

[238]  A. Tandon,et al.  A technique for retrieving antigens in formalin-fixed, routinely acid-decalcified, celloidin-embedded human temporal bone sections for immunohistochemistry. , 1992, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[239]  H. Merz,et al.  Constant Detection of CD2, CD3, CD4, and CD5 in Fixed and Paraffin-embedded Tissue Using the Peroxidase-mediated Deposition of Biotin-Tyramide , 1997, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[240]  Clive R. Taylor,et al.  Standardization of immunohistochemistry based on antigen retrieval technique for routine formalin-fixed tissue sections , 1998 .

[241]  T. Iwaki,et al.  Hydrated autoclave pretreatment enhances tau immunoreactivity in formalin-fixed normal and Alzheimer's disease brain tissues. , 1991, Laboratory investigation; a journal of technical methods and pathology.

[242]  P. Lukinmaa,et al.  Immunohistochemical Localization of Types I, V, and VI Collagen in Human Permanent Teeth and Periodontal Ligament , 1992, Journal of dental research.

[243]  Susan Van Noorden,et al.  Failure of Ultrasonic Vibration as a Means of Antigen Retrieval in Routine Diagnostic Immunocytochemistry , 2000, Applied immunohistochemistry & molecular morphology (Print).

[244]  A. Habeeb,et al.  Reaction of proteins with glutaraldehyde. , 1968, Archives of biochemistry and biophysics.

[245]  K. Cooper,et al.  Beware of biotin: a source of false-positive immunohistochemistry , 2001 .

[246]  K. Kashima,et al.  Ultrastructure and chromogranin A immunogold labelling of ECL cell carcinoids , 2005, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[247]  T. Seidal,et al.  Interpretation and Quantification of Immunostains , 2001, The American journal of surgical pathology.

[248]  O. Suhr,et al.  Evaluation of immunohistochemical staining of human duodenal endocrine cells after microwave antigen retrieval , 1997, The Histochemical Journal.

[249]  R. Cote,et al.  Antigen retrieval immunohistochemistry under the influence of pH using monoclonal antibodies. , 1995, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[250]  D. Arber Effect of Prolonged Formalin Fixation on the Immunohistochemical Reactivity of Breast Markers , 2002, Applied immunohistochemistry & molecular morphology : AIMM.

[251]  R. Burry Electron Microscopic Immunocytochemistry , 2010 .

[252]  J. Buxbaum,et al.  Pepsin pretreatment allows collagen IV immunostaining of blood vessels in adult mouse brain , 2007, Journal of Neuroscience Methods.

[253]  A. Chan,et al.  The contribution of bifunctional SkipDewax pretreatment solution, rabbit monoclonal antibodies, and polymer detection systems in immunohistochemistry. , 2007, Archives of pathology & laboratory medicine.

[254]  A. Leong,et al.  Newer developments in immunohistology , 2006, Journal of Clinical Pathology.

[255]  F. Bianchi,et al.  Immunohistochemical demonstration of the small GTPase RhoA on epoxy-resin embedded sections. , 2004, Micron.