86 results on '"Shan-Rong Shi"'
Search Results
2. Application of Immunohistochemistry in Cytology
- Author
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Yan Shi, Melissa Yee-Chang, and Shan-Rong Shi
- Subjects
Medical Laboratory Technology ,Histology ,Pathology and Forensic Medicine - Published
- 2022
3. Commentary on 'Antigen Retrieval Immunohistochemistry: Past, Present, and Future'
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Clive R. Taylor and Shan-Rong Shi
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Histology ,Anatomy - Published
- 2022
4. Antigen Retrieval Immunohistochemistry Based Research and Diagnostics
- Author
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Shan-Rong Shi, Clive R. Taylor, Shan-Rong Shi, Clive R. Taylor and Shan-Rong Shi, Clive R. Taylor, Shan-Rong Shi, Clive R. Taylor
- Published
- 2011
5. New Dimensions of Antigen Retrieval Technique: 28 Years of Development, Practice, and Expansion
- Author
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Shan-Rong Shi, Jiang Gu, Clive R. Taylor, and Yan Shi
- Subjects
0301 basic medicine ,Proteomics ,Histology ,Tissue Fixation ,Formalin fixed paraffin embedded ,Computer science ,Computational biology ,Kidney ,Mass Spectrometry ,Pathology and Forensic Medicine ,Heating ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Animals ,Humans ,Paraffin embedding ,Antigens ,Paraffin Embedding ,Extramural ,Kidney pathology ,Kidney metabolism ,Immunohistochemistry ,Medical Laboratory Technology ,030104 developmental biology ,Antigen retrieval ,chemistry ,030220 oncology & carcinogenesis - Abstract
This review article summarized recent advances in the heat-induced antigen retrieval technique with numerous scientific fields in addition to immunohistochemistry. Particularly, proteomics including imaging mass spectrometry, extraction of proteins from formalin-fixed, paraffin-embedded (FFPE) tissues. Some novel approaches such as FFPE tissue-based renal immunopathology based on modified double heating protocols are also introduced in this review for further development. In general, the FFPE tissue housed in pathology worldwide is an invaluable treasure, and the simple method of heat-induced antigen retrieval is the gold key to open the door of this treasure.
- Published
- 2019
6. Protein Extraction from Formalin-fixed, Paraffin-embedded Tissue Sections: Quality Evaluation by Mass Spectrometry
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Cheng Lee, Clive R. Taylor, Cheng Liu, Shan-Rong Shi, and Brian M. Balgley
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Quality Control ,Histology ,Formalin fixed paraffin embedded ,Mass spectrometry ,Mass Spectrometry ,Heating ,Fixatives ,Mice ,chemistry.chemical_compound ,Fresh Tissue ,Formaldehyde ,Protein purification ,Animals ,Humans ,Mammary Glands, Human ,Gel electrophoresis ,Paraffin Embedding ,Chromatography ,Extraction (chemistry) ,Proteins ,Hydrogen-Ion Concentration ,Radioimmunoprecipitation Assay ,Molecular biology ,Kidney Neoplasms ,Liver ,Antigen retrieval ,chemistry ,Electrophoresis, Polyacrylamide Gel ,Anatomy - Abstract
A satisfactory protocol of protein extraction has been established based on the heat-induced antigen retrieval (AR) technique widely applied in immunohistochemistry for archival formalin-fixed, paraffin-embedded (FFPE) tissue sections. Based on AR, an initial serial experiment to identify an optimal protocol of heat-induced protein extraction was carried out using FFPE mouse tissues. The optimal protocol for extraction of proteins was then performed on an archival FFPE tissue of human renal carcinoma. FFPE sections were boiled in a retrieval solution of Tris-HCl containing 2% SDS, followed by incubation. Fresh tissue taken from the same case of renal carcinoma was processed for extraction of proteins by a conventional method using radioimmunoprecipitation assay solution, to compare the efficiency of protein extraction from FFPE tissue sections with extraction from fresh tissue. As a control, further sections of the same FFPE sample were processed by the same procedure without heating treatment. Evaluation of the quality of protein extracted from FFPE tissue was done using gel electrophoresis and mass spectrometry, showing most identified proteins extracted from FFPE tissue sections were overlapped with those extracted from fresh tissue.
- Published
- 2006
7. Protein-embedding Technique: A Potential Approach to Standardization of Immunohistochemistry for Formalin-fixed, Paraffin-embedded Tissue Sections
- Author
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Jeanette Perez, Shan-Rong Shi, Clive R. Taylor, and Cheng Liu
- Subjects
Histology ,Materials science ,Formalin fixed paraffin embedded ,Polymers ,Microsphere ,law.invention ,Matrix (chemical analysis) ,Fixatives ,Mice ,chemistry.chemical_compound ,law ,Formaldehyde ,Microtome ,Animals ,Horses ,Paraffin Embedding ,Goats ,S100 Proteins ,Antibodies, Monoclonal ,Proteins ,Microtomy ,Reference Standards ,Immunohistochemistry ,Molecular biology ,Microspheres ,Tissue sections ,Antigen retrieval ,chemistry ,Keratins ,Embedding ,Anatomy ,Biomedical engineering - Abstract
A serial study was performed to develop a protein-embedding technique for standardization of immunohistochemistry (IHC) on formalin-fixed, paraffin-embedded (FFPE) tissue sections. A protein carrier matrix must have two phases, a liquid phase to allow uniform mixing of a protein and a solid phase forming a ‘block’ that can be fixed and processed in the same manner as human tissue. This standardized protein block would serve as a source of thin sections for control of IHC and therefore must also withstand the boiling conditions of antigen retrieval (AR). After multiple experiments, a method was developed utilizing polymer microsphere (beads) as a support medium for protein. The method showed particular promise for quantitative IHC.
- Published
- 2005
8. Hyperphosphorylation of pRb: a mechanism for RB tumour suppressor pathway inactivation in bladder cancer
- Author
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Peter A. Jones, Carlos Cordon-Cardo, Peter J. Goebell, Yuen Kai Fung, Mohammad Alavi-Tafreshi, Shan Rong Shi, Ben George, Sunanda J. Chatterjee, Ram H. Datar, and Richard J. Cote
- Subjects
Tumor suppressor gene ,Cell growth ,Blotting, Western ,Hyperphosphorylation ,Transfection ,Biology ,Retinoblastoma Protein ,Neoplasm Proteins ,Pathology and Forensic Medicine ,Immunoenzyme Techniques ,Blot ,Loss of heterozygosity ,Cyclin D1 ,Urinary Bladder Neoplasms ,Image Processing, Computer-Assisted ,Tumor Cells, Cultured ,Cancer research ,Humans ,Phosphorylation ,biological phenomena, cell phenomena, and immunity ,neoplasms ,Cell Division ,Cyclin-Dependent Kinase Inhibitor p16 - Abstract
Loss of heterozygosity, mutations or deletions of the RB1 gene usually result in loss of pRb expression, which has been regarded as an indicator of loss of pRb function in human tumours. It has previously been shown that in addition to loss of pRb expression, aberrantly high (pRb2+) pRb expression also indicates loss of pRb function in bladder tumours compared with moderate (normal, pRb1+) pRb expression. The aim of this study was to elucidate the mechanism by which pRb is functionally inactivated in bladder tumours expressing aberrantly high levels of pRb. Constitutive phosphorylation was therefore investigated as a mechanism of pRb inactivation in bladder tumours. Of 28 bladder tumours examined, western blotting demonstrated pRb hyperphosphorylation in 5/7 (71%) pRb2+ bladder tumours compared with only 4/11 (36%) pRb1+ tumours (p = 0.002). All cases with undetectable pRb showed moderate to high p16 expression and none showed cyclin D1 expression by immunohistochemistry. All pRb1+ tumours with underphosphorylated pRb showed p16 but not cyclin D1 expression. All pRb2+ tumours with hyperphosphorylated pRb showed loss of p16 expression and/or cyclin D1 overexpression. Thus, elevated pRb expression was associated with pRb hyperphosphorylation, which, in turn, was associated with loss of p16 expression and/or increased cyclin D1 expression. In order to analyse this association in vitro, T24 cells, which express high levels of pRb, were transfected with p16 cDNA. Transfection with p16 cDNA resulted in a marked decrease in pRb phosphorylation, decreased cell proliferation, and a change in expression of pRb from high to moderate phenotype as assessed by immunohistochemistry. This paper gives the biological basis for constitutive alteration of pRb function in human tumours in the presence of an intact, expressed pRb protein; the mechanism of pRb inactivation is through hyperphosphorylation, which results from loss of p16 expression and/or cyclin D1 overexpression. Immunohistochemical expression of pRb appears to be a reliable indicator of pRb function.
- Published
- 2004
9. Combined Effects of p53, p21, and pRb Expression in the Progression of Bladder Transitional Cell Carcinoma
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Donald G. Skinner, Peter J. Goebell, Susan Groshen, Richard J. Cote, John P. Stein, Ram H. Datar, Shan Rong Shi, Ben George, Sunanda J. Chatterjee, Conway Gee, Lillian L. Young, and David Youssefzadeh
- Subjects
Cancer Research ,medicine.medical_specialty ,Pathology ,Urinary bladder ,business.industry ,Proportional hazards model ,medicine.medical_treatment ,Urinary system ,Urology ,medicine.disease ,Cystectomy ,medicine.anatomical_structure ,Transitional cell carcinoma ,Oncology ,Carcinoma ,medicine ,Lymph ,business ,Survival analysis - Abstract
Purpose To determine the combined effects of p53, p21, and pRb alterations in predicting the progression of bladder transitional cell carcinoma. Patients and Methods p53, p21, and pRb expression was examined immunohistochemically on archival radical cystectomy samples from 164 patients with invasive or high-grade recurrent superficial transitional cell carcinoma (TCC; lymph node–negative, 117 patients; lymph node–positive, 47 patients). Median follow-up was 8.6 years. Based on percentage of nuclear reactivity, p53 was considered as wild-type (0% to 10%) or altered (> 10%); p21 was scored as wild-type (>10%) or altered (< 10%); and pRb status was considered wild-type (1% to 50%) or altered (0% or > 50%). Results As individual determinants, the p53, p21, and pRb status were independent predictors of time to recurrence (P < .001, P < .001, and P < .001, respectively), and overall survival (P < .001, P = .002, and P = .001, respectively). By examining these determinants in combination, patients were categorized as group I (no alteration in any determinant, 47 patients), group II (any one determinant altered, 51 patients), group III (any two determinants altered, 42 patients), and group IV (all three determinants altered, 24 patients). The 5-year recurrence rates in these groups were 23%, 32%, 57%, and 93%, respectively (log-rank P < .001), and the 5-year survival rates were 70%, 58%, 33%, and 8%, respectively (log-rank P < .001). After stratifying by stage, the number of altered proteins remained significantly associated with time to recurrence and overall survival. Conclusion This study suggests that alterations in p53, p21, and pRb act in cooperative or synergistic ways to promote bladder cancer progression. Examining these determinants in combination provides additional information above the use of a single determinant alone.
- Published
- 2004
10. Antigen retrieval immunohistochemistry used for routinely processed celloidin-embedded human temporal bone sections: standardization and development
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Shan Rong Shi, Richard J. Cote, and Clive R. Taylor
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Pathology ,medicine.medical_specialty ,Tissue Fixation ,Standardization ,computer.software_genre ,Sensitivity and Specificity ,chemistry.chemical_compound ,Molecular level ,Temporal bone ,Humans ,Medicine ,Ear Diseases ,Molecular Biology ,business.industry ,Collodion ,Reproducibility of Results ,Temporal Bone ,General Medicine ,Immunohistochemistry ,Otorhinolaryngology ,Antigen retrieval ,chemistry ,Ear, Inner ,Tissue Adhesives ,Surgery ,Artificial intelligence ,business ,computer ,Natural language processing - Abstract
The use of immunohistochemistry (IHC) in routinely processed celloidin-embedded human temporal bone section has created a fruitful field in understanding the pathogenesis and pathophysiology of the human inner ear at a molecular level since the early 1990s when the antigen retrieval (AR) technique was developed. This review article focuses on several critical technical issues of AR technique based predominantly on our experiences and suggestions concerning further development and standardization of AR-IHC for IHC study of human temporal bone section, as well as other tissues embedded in celloidin. Examples of using AR-IHC in human temporal bone sections collected include our unpublished data in order to indicate the potential utility of this novel method. Suggestions of further development of AR techniques are proposed for references and comments.
- Published
- 1998
11. Standardization of Immunohistochemistry Based on Antigen Retrieval Technique for Routine Formalin-fixed Tissue Sections
- Author
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Benjaporn Chaiwun, Taiying Chen, Yan Shi, Shan Rong Shi, Clive R. Taylor, Richard J. Cote, Debra Hawes, and Lillian L. Young
- Subjects
Medical Laboratory Technology ,chemistry.chemical_compound ,Pathology ,medicine.medical_specialty ,Tissue sections ,Antigen retrieval ,chemistry ,medicine ,Immunohistochemistry ,Formalin fixed ,Anatomy - Published
- 1998
12. Antigen Retrieval Immunohistochemistry: Practice and Development
- Author
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Lillian L. Young, Clive R. Taylor, Shan Rong Shi, and Richard J. Cote
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Medical Laboratory Technology ,chemistry.chemical_compound ,Histology ,Antigen retrieval ,chemistry ,business.industry ,Medicine ,Anatomy ,Bioinformatics ,business ,Data science ,Review article - Abstract
The antigen retrieval (AR) technique is a novel, simple, and reliable approach to routine immunohistochemistry (IHC) developed in 1991. It has increasingly been used in pathology and analytical morphology, with a wide fruitful field opened for both clinical and research projects. Hundreds of articles concerning AR-IHC have been published in a variety of journals all over the world. As a recently developed technique, the wide use of AR has raised numerous questions for further study. This review article focuses on several key points of the method and further developments of the AR technique that are different from some review articles published recently. (The J Histotechnol 20:145, 1997)
- Published
- 1997
13. Immunohistochemical detection of thrombospondin-1 in formalin-fixed, paraffin-embedded tissue
- Author
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Gary D. Grossfeld, Shan Rong Shi, Donald G. Skinner, Clive R. Taylor, David A. Ginsberg, Kathryn A. Rich, and Richard J. Cote
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endocrine system ,Pathology ,medicine.medical_specialty ,Tissue Fixation ,Histology ,Angiogenesis ,Metastasis ,Heating ,Immunoenzyme Techniques ,chemistry.chemical_compound ,immune system diseases ,Formaldehyde ,Thrombospondin 1 ,medicine ,Frozen Sections ,Microwaves ,Carcinoma, Transitional Cell ,Thrombospondin ,Membrane Glycoproteins ,Paraffin Embedding ,Tissue microarray ,Staining and Labeling ,virus diseases ,medicine.disease ,Transitional cell carcinoma ,Urinary Bladder Neoplasms ,Antigen retrieval ,chemistry ,Immunohistochemistry ,Anatomy ,Thrombospondins - Abstract
Thrombospondin-1 (TSP) is a 450-KD glycoprotein that was initially discovered in the platelet alpha-granule. It now appears that TSP is intimately involved in the regulation of a variety of cellular functions and cell-to-cell interactions. Recently, it has been demonstrated that TSP functions as a p53-dependent inhibitor of angiogenesis in cultured fibroblasts from Li-Fraumeni patients and therefore may be an important factor involved with tumor invasion and metastasis. It has previously been demonstrated that TSP can be detected in frozen tissue sections by immunohistochemical methods. Our objective in this study was to determine the optimal antigen retrieval (AR) protocol for detection of TSP in formalin-fixed, paraffin-embedded tissue by using tissue sections from patients with invasive transitional cell carcinoma of the bladder. The optimal AR protocol was determined utilizing a variety of heating conditions and antigen retrieval buffers. Our results demonstrate that TSP can be reliably detected in paraffin-embedded tissue by immunohistochemical techniques that utilize AR with high-temperature microwave heating and a low-pH Tris-HCI buffer. The importance of this method is that it allows the reliable detection of TSP in archival tissue. This should facilitate further investigation into TSP's role in the regulation of cellular processes, including its influence on tumor angiogenesis and metastasis.
- Published
- 1996
14. 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
- Author
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Christina Yang, Hong Ji Xu, William F. Benedict, Chen Chen, Richard J. Cote, Clive R. Taylor, and Shan Rong Shi
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Frozen section procedure ,Pathology ,medicine.medical_specialty ,biology ,Chemistry ,Retinoblastoma ,Retinoblastoma protein ,medicine.disease ,Pathology and Forensic Medicine ,Staining ,chemistry.chemical_compound ,Antigen retrieval ,medicine ,biology.protein ,Carcinoma ,Immunohistochemistry ,Immunostaining - Abstract
The retinoblastoma (RB) gene, which encodes the nuclear RB protein (pRB), is believed to be involved in cell cycle control and cell differentiation. Studies have demonstrated that loss of RB function may play a role in tumour formation and progression of a variety of human tumours, such as bladder, lung, breast, and prostate cancers. The immunohistochemical detection of pRB expression in formalin-paraffin sections of human cancer has potential advantages of convenience, economy, and compatibility with routine surgical pathology practice. In practice, however, results using pRB antibodies on routinely processed, paraffin-embedded tissue have been inconsistent. In this study, the antigen retrieval (AR) method has been applied to the immunohistochemical detection of pRB in paraffin-embedded tissues and a 'test battery' approach has been developed to identify the principal variables that result in the optimal AR protocol. This approach includes the use of buffered solutions at pH 1, 6, and 10 with three different heating conditions (temperatures 120 degrees C, 100 degrees C, and 90 degrees C). In the example described here with antibody RB-WL-1, the low pH solution with the microwave heating at 100 degrees C proved most effective. Both fresh and routinely processed formalin-paraffin tissues of normal and bladder carcinoma were used for a comparison of the pRB immunostaining. The AR method was evaluated by comparing the immunohistochemical staining result on routinely processed formalin-paraffin sections with frozen sections of the same tumour. A consistent intensity of immunohistochemical staining for pRB was achieved using the identified optimal AR protocol on formalin-paraffin sections. All slides showed positive staining of pRB in normal mesenchymal and epithelial tissues. The pattern of pRB localization and intensity of staining was similar to that obtained in frozen sections, though the intensity obtained by AR treatment on paraffin sections was slightly to moderately stronger than that obtained in frozen sections. Once the protocol was identified, it was tested using routinely processed paraffin tissue sections of 245 cases of bladder carcinoma, with consistent pRB immunostaining results. The protocol described is simple to perform and gives reproducible results for evaluation of pRB expression by immunohistochemistry.
- Published
- 1996
15. Comparative Study of Antigen Retrieval Heating Methods: Microwave, Microwave and Pressure Cooker, Autoclave, and Steamer
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Chen Chen, Lillian L. Young, Richard J. Cote, Shan Rong Shi, Christina Yang, and Clive R. Taylor
- Subjects
Male ,Histology ,Staining and Labeling ,Chemistry ,Microwave oven ,Analytical chemistry ,Cooker ,General Medicine ,Formalin fixed ,Autoclave ,Dilution ,Heating ,Medical Laboratory Technology ,chemistry.chemical_compound ,Antigen retrieval ,Humans ,Female ,Antigens ,Microwaves ,Immunostaining ,Microwave - Abstract
We present a study comparing the most popular heating methods currently used for antigen retrieval (AR) immunostaining: the microwave oven, microwave with pressure cooker, autoclave, and steamer heating. A panel of 21 antibodies was tested on formalin fixed, paraffin embedded sections using these heating methods and Tris-HC1 buffer, pH 9.5, plus 5% urea as the AR solution. Three observers independently evaluated the intensity of AR immunostaining. All heating methods yielded good results for AR immunostaining. There were only minor differences among the heating methods for AR when the optimal concentration of primary antibody for normal immunostaining was used; however, background staining may occasionally be troublesome if antibodies are not retitrated and diluted further for use on tissues after AR. Significant differences were observed only after further dilution of the primary antibodies: the microwave pressure cooker, extended microwave heating (5 min x 4) and autoclave heating then showed a similar intensity of staining that was stronger than results obtained with the steamer (20 min) or regular microwave heating (5 min x 2). Extension of the steamer heating time, however, yielded equivalent results. This study indicates that different heating methods can yield similar intensities of AR immunostaining if the heating times are adjusted appropriately. It is noteworthy that, in general, the adjusted conditions for maximal retrieval differ from those most widely cited in the literature, or recommended by manufacturers. That several heating devices may provide similar results permits the use of different AR heating methods according to the equipment available. This study also is an early step in standardizing the AR immunostaining protocol by providing uniform conditions for "maximal retrieval" as a common end point for all laboratories.
- Published
- 1996
16. Detection of Occult Bone Marrow Micrometastases in Patients with Operable Lung Carcinoma
- Author
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Shan Rong Shi, Clive R. Taylor, Richard J. Cote, Benjaporn Chaiwun, James Harvey, Andy Sherrod, Su Chiu Chen, Edward J. Beattie, and Susan Groshen
- Subjects
medicine.medical_specialty ,Systemic disease ,Pathology ,Lung Neoplasms ,Gastroenterology ,Epithelium ,Metastasis ,Antigens, Neoplasm ,Risk Factors ,Carcinoma, Non-Small-Cell Lung ,Internal medicine ,medicine ,Adjuvant therapy ,Carcinoma ,Humans ,Stage (cooking) ,Lung cancer ,business.industry ,Respiratory disease ,Antibodies, Monoclonal ,medicine.disease ,Immunohistochemistry ,Survival Rate ,medicine.anatomical_structure ,Keratins ,Surgery ,Bone marrow ,Neoplasm Recurrence, Local ,Bone Marrow Neoplasms ,business ,Research Article - Abstract
OBJECTIVES: A large proportion of patients with operable lung carcinoma (no evidence of systemic spread of tumor) develop metastatic disease after primary therapy. More sensitive and specific methods are needed to identify patients at highest risk for recurrence who may benefit most from adjuvant therapy, while sparing those patients who do not require such treatment. SUMMARY BACKGROUND DATA: Using epithelial-specific monoclonal antibodies, the authors have developed an immunocytochemical assay capable of detecting as few as 2 lung cancer cells in 1 million bone marrow cells. METHODS: The assay was used to test the bone marrow (from resected ribs) of 43 patients with primary non-small cell lung carcinoma who showed no clinical or pathologic evidence of systemic disease. RESULTS: Occult bone marrow micrometastases (BMMs) were detected in 40% of patients (17/43) with non-small cell lung cancer, including 29% (5/17) of patients with stage I or II disease and 46% of whom (12/26) had stage III disease. The median follow-up was 13.6 months. Patients with occult BMMs had significantly shorter times to disease recurrence compared with patients without BMMs (7.3 vs. > 35.1 months, p = 0.0009). Furthermore, for patients with stage I or II disease, the presence of occult BMMs was significantly associated with a higher rate of recurrence (p = 0.0004). CONCLUSIONS: The detection of occult BMMs identifies patients with operable non-small cell lung carcinoma who are at significantly increased risk for recurrence, independent of tumor stage, and may be useful in evaluating patients for adjuvant treatment protocols.
- Published
- 1995
17. Antigen retrieval immunohistochemistry under the influence of pH using monoclonal antibodies
- Author
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S A Imam, Shan Rong Shi, Richard J. Cote, Clive R. Taylor, and Lillian L. Young
- Subjects
Cytoplasm ,Hot Temperature ,Histology ,medicine.drug_class ,Antibodies, Monoclonal ,Buffer solution ,Buffers ,Hydrogen-Ion Concentration ,Monoclonal antibody ,Immunohistochemistry ,Molecular biology ,Staining ,chemistry.chemical_compound ,chemistry ,Antigen retrieval ,Antigen ,Antigens, Surface ,medicine ,Humans ,Antigens ,Anatomy ,Artifacts ,Sodium acetate ,Immunostaining - Abstract
Antigen retrieval (AR) incorporating high-temperature microwave (MW) heating of tissue sections before immunostaining is a revolutionary technique that can unmask the antigens in formalin-fixed tissue sections, thus making them available for immunohistochemical staining. Although high temperature is believed to be the primary mechanism in retrieval of antigens, a variety of chemical solutions have been tested to define an optimal AR solution. We tested the hypothesis that pH of the AR solution may influence the quality of immunostaining by using seven different AR buffer solutions at a series of different pH values ranging from 1 to 10. We evaluated the staining of monoclonal antibodies to cytoplasmic antigens (AE1, HMB45, NSE), nuclear antigens (MIB-1, PCNA, ER), and cell surface antigens (MT1, L26, EMA) on routinely formalin-fixed, paraffin-embedded sections under different pH conditions with MW heating for 10 min. The intensity of immunostaining was graded in a blinded fashion. The pH value of the AR buffer solution was carefully measured before, immediately after, and 15 min after the AR procedure. The influence of pH on AR immunohistochemical staining can be summarized into three patterns. Some antigens (L26, PCNA, AE1, EMA, and NSE) showed excellent retrieval throughout the pH range. Other antigens (MIB1 and ER) showed strong intensity of immunohistochemical staining at very low pH and at neutral to high pH, but a dramatic decrease in the intensity of the AR immunostaining at moderately acidic pH (pH 3-6). Still others (MT1 and HMB45) showed increasing intensity of the AR immunostaining with increasing pH, but only weak immunostaining at low pH. Among the seven buffer solutions at any given pH value, the intensity of AR immunostaining was very similar. However, Tris-HCl buffer tended to produce better results at higher pH, compared with other buffers. Although high-temperature heating is believed to be the most important factor for the AR technique, the pH value of the AR solution is an important co-factor for some antigens. Optimization of the AR system should therefore include optimization of the pH of the AR solution. Our results indicate that AR immunostaining of Tris-HCl or sodium acetate buffer at pH 8-9 may be suitable for most antigens, although certain nuclear antigens show optimal staining at low pH.
- Published
- 1995
18. Complete solubilization of formalin-fixed, paraffin-embedded tissue may improve proteomic studies
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Jeffrey T. Mason, Shan-Rong Shi, Carol B. Fowler, and Clive R. Taylor
- Subjects
Paraffin Embedding ,Formalin fixed paraffin embedded ,Proteome ,business.industry ,Clinical Biochemistry ,Reproducibility of Results ,Computational biology ,Biology ,Bioinformatics ,Proteomics ,Article ,chemistry.chemical_compound ,Fixatives ,Antigen retrieval ,chemistry ,Solubilization ,Paraffin ,Tissue proteomics ,Tissue bank ,Formaldehyde ,Humans ,Personalized medicine ,business ,Biomarkers - Abstract
Tissue-based proteomic approaches (tissue proteomics) are essential for discovering and evaluating biomarkers for personalized medicine. In any proteomics study, the most critical issue is sample extraction and preparation. This problem is especially difficult when recovering proteins from formalin-fixed, paraffin-embedded (FFPE) tissue sections. However, improving and standardizing protein extraction from FFPE tissue is a critical need because of the millions of archival FFPE tissues available in tissue banks worldwide. Recent progress in the application of heat-induced antigen retrieval (AR) principles for protein extraction from FFPE tissue has resulted in a number of published FFPE tissue proteomics studies. However, there is currently no consensus on the optimal protocol for protein extraction from FFPE tissue or accepted standards for quantitative evaluation of the extracts. Standardization is critical to ensure the accurate evaluation of FFPE protein extracts by proteomic methods such as reverse phase protein arrays (RPPA), which is now in clinical use. In our view, complete solubilization of FFPE tissue samples is the best way to achieve the goal of standardizing the recovery of proteins from FFPE tissues. However, further studies are recommended to develop standardized protein extraction methods to ensure quantitative and qualitative reproducibility in the recovery of proteins from FFPE tissues.
- Published
- 2012
19. Antigen retrieval immunohistochemistry: review and future prospects in research and diagnosis over two decades
- Author
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Shan-Rong Shi, Yan Shi, and Clive R. Taylor
- Subjects
Proteomics ,Histology ,Tissue Fixation ,Formalin fixed paraffin embedded ,Reviews ,Translational research ,Biology ,Bioinformatics ,chemistry.chemical_compound ,Diagnosis ,Animals ,Humans ,Antigens ,Reference standards ,business.industry ,Reference Standards ,Data science ,Immunohistochemistry ,Antigen retrieval ,chemistry ,Tissue proteomics ,Archival tissue ,Personalized medicine ,Anatomy ,business - Abstract
As a review for the 20th anniversary of publishing the antigen retrieval (AR) technique in this journal, the authors intend briefly to summarize developments in AR-immunohistochemistry (IHC)–based research and diagnostics, with particular emphasis on current challenges and future research directions. Over the past 20 years, the efforts of many different investigators have coalesced in extending the AR approach to all areas of anatomic pathology diagnosis and research and further have led to AR-based protein extraction techniques and tissue-based proteomics. As a result, formalin-fixed paraffin-embedded (FFPE) archival tissue collections are now seen as a literal treasure of materials for clinical and translational research to an extent unimaginable just two decades ago. Further research in AR-IHC is likely to focus on tissue proteomics, developing a more efficient protocol for protein extraction from FFPE tissue based on the AR principle, and combining the proteomics approach with AR-IHC to establish a practical, sophisticated platform for identifying and using biomarkers in personalized medicine.
- Published
- 2011
20. Techniques of Immunohistochemistry
- Author
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Nancy J. Barr, Shan-Rong Shi, and Clive R. Taylor
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medicine.medical_specialty ,Standardization ,Chemistry ,medicine ,Immunohistochemistry ,Medical physics - Published
- 2011
21. Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections
- Author
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Benjaporn Chaiwun, Shan-Rong Shi, Richard J. Cote, Clive R. Taylor, and Lillian L. Young
- Subjects
Male ,Tissue Fixation ,Histology ,Pronase ,Buffers ,Citric Acid ,Immunoenzyme Techniques ,chemistry.chemical_compound ,Antigen ,Antigens, Neoplasm ,medicine ,Frozen Sections ,Humans ,Urea ,Citrates ,Paraffin Embedding ,Chromatography ,Staining and Labeling ,Chemistry ,Carcinoma ,Prostate ,Prostatic Neoplasms ,Buffer solution ,Trypsin ,Staining ,Biochemistry ,Antigen retrieval ,Receptors, Androgen ,Anatomy ,Citric acid ,Immunostaining ,medicine.drug - Abstract
We developed a staining protocol for demonstration of androgen receptor (AR) in formalin-fixed, paraffin-embedded tissue sections. The method is based on the antigen retrieval microwave (MW) heating technique. Results are compared with different types of enzyme digestion pre-treatments. The strongest immunostaining signal and clearest background were obtained by MW heating of dewaxed paraffin sections in 5% urea or citrate buffer solution (pH 6); pure distilled water gave less consistent results. Enzymatic digestion with pepsin (0.05% in 2 N HCl) for 30 min at room temperature, or trypsin followed by pronase, or pronase digestion alone, also produced enhanced staining of AR in some cases, but there was more nonspecific background, and specific reactivity was less intense. The antigen retrieval MW method can be used to demonstrate AR epitope in prostate tissue after fixation in formalin for as long as 7 days. AR immunolocalization was also compared in frozen and paraffin sections processed from the same specimen of prostate carcinoma tissue and was found to be qualitatively and quantitatively similar. This study also provided new information concerning the basic principles of the antigen retrieval MW method that may be helpful in further development of this technique.
- Published
- 1993
22. Extended Application of Antigen Retrieval Technique in Immunohistochemistry and in SITU Hybridization
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Shan-Rong Shi and Clive R. Taylor
- Subjects
chemistry.chemical_compound ,Pathology ,medicine.medical_specialty ,Antigen retrieval ,chemistry ,medicine ,Immunohistochemistry ,In situ hybridization - Published
- 2010
23. Standardization of Antigen Retrieval Techniques Based on the Test Battery Approach
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Clive R. Taylor and Shan-Rong Shi
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Test battery ,chemistry.chemical_compound ,Standardization ,Antigen retrieval ,chemistry ,Computer science ,Speech recognition - Published
- 2010
24. Standardization of Immunohistochemistry Based on Antigen Retrieval Technique
- Author
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Clive R. Taylor and Shan-Rong Shi
- Subjects
chemistry.chemical_compound ,Information retrieval ,Antigen retrieval ,chemistry ,Standardization ,business.industry ,Immunohistochemistry ,Medicine ,business - Published
- 2010
25. Tissue Cell Sample Preparation: Lessons from the Antigen Retrieval Technique
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Clive R. Taylor and Shan-Rong Shi
- Subjects
chemistry.chemical_compound ,Pathology ,medicine.medical_specialty ,Antigen retrieval ,chemistry ,medicine ,Tissue cell ,Sample preparation ,Biology - Published
- 2010
26. Extraction of DNA/RNA from Formalin-Fixed, Paraffin-Embedded Tissue Based on the Antigen Retrieval Principle
- Author
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Shan-Rong Shi and Clive R. Taylor
- Subjects
chemistry.chemical_compound ,Formalin fixed paraffin embedded ,Antigen retrieval ,Chemistry ,Extraction (chemistry) ,Molecular biology - Published
- 2010
27. Key Issues and Strategies of Standardization for Quantifiable Immunohistochemistry
- Author
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Kevin A. Roth, Clive R. Taylor, and Shan-Rong Shi
- Subjects
Engineering drawing ,Engineering ,Standardization ,business.industry ,Engineering ethics ,business ,Key issues - Published
- 2010
28. Symbiosis of Immunohistochemistry and Proteomics: Marching to a New ERA
- Author
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Clive R. Taylor, Shan-Rong Shi, and Brian M. Balgley
- Subjects
Chromatography ,Symbiosis ,Chemistry ,Computational biology ,Proteomics - Published
- 2010
29. Antigen Retrieval Immunohistochemistry Based Research and Diagnostics
- Author
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Shan-Rong Shi and Clive R. Taylor
- Subjects
chemistry.chemical_compound ,Pathology ,medicine.medical_specialty ,Antigen retrieval ,chemistry ,business.industry ,Immunology ,Immunohistochemistry ,Medicine ,business - Published
- 2010
30. Standard Reference Material: Proteinembedding Technique and Design of Bar Code
- Author
-
Jiang Gu, Clive R. Taylor, and Shan-Rong Shi
- Subjects
Engineering drawing ,Engineering ,business.industry ,Bar (music) ,Code (cryptography) ,business - Published
- 2010
31. Appendix: Related Laboratory Protocols
- Author
-
Clive R. Taylor and Shan-Rong Shi
- Subjects
medicine.medical_specialty ,medicine.anatomical_structure ,Computer science ,medicine ,Medical physics ,Appendix - Published
- 2010
32. CONTRIBUTORS
- Author
-
N. Volkan Adsay, Nancy J. Barr, Olca Basturk, Parul Bhargava, Rohit Bhargava, Mamatha Chivukula, Cheryl M. Coffin, Jessica M. Comstock, David J. Dabbs, Sanja Dacic, Ronald A. DeLellis, Jonathan I. Epstein, Nicole N. Esposito, Eduardo J. Ezyaguirre, Alton B. Farris III, Jeffrey D. Goldsmith, Samuel P. Hammar, Jason L. Hornick, Jennifer L. Hunt, Marshall E. Kadin, Alyssa M. Krasinskas, Alvin W. Martin, Paul E. McKeever, George J. Netto, Yuri E. Nikiforov, Marina N. Nikiforova, James W. Patterson, Joseph T. Rabban, Shan-Rong Shi, Sandra J. Shin, Robert A. Soslow, Paul E. Swanson, Clive R. Taylor, Diana O. Treaba, David H. Walker, Jeremy C. Wallentine, Mark R. Wick, Sherif R. Zaki, and Charles Z. Zaloudek
- Published
- 2010
33. S-100 Protein in Human Inner Ear: Use of a Novel Immunohistochemical Technique on Routinely Processed, Celloidin- Embedded Human Temporal Bone Sections
- Author
-
Atul K. Tandon, Shan-Rong Shi, Krishan L. Kalra, and Catherine Coté
- Subjects
Pathology ,medicine.medical_specialty ,Spiral limbus ,Biology ,chemistry.chemical_compound ,Temporal bone ,otorhinolaryngologic diseases ,medicine ,Humans ,Inner ear ,Cochlea ,Vestibular system ,Tissue Embedding ,Histocytochemistry ,S100 Proteins ,Collodion ,Temporal Bone ,Anatomy ,Facial nerve ,medicine.anatomical_structure ,Otorhinolaryngology ,Antigen retrieval ,chemistry ,Ear, Inner ,Spiral ligament ,sense organs - Abstract
The authors recently developed a new antigen retrieval technique which allows immunohistochemical detection of proteins in routinely processed, celloidin-embedded human temporal bone sections. This technique was used in the present study, which reports the occurrence of S-100 protein in the human inner ear. Fifteen celloidin-embedded human temporal bone sections, obtained from the Eastern Temporal Bone Bank at the Massachusetts Eye and Ear Infirmary, were tested with the monoclonal antibody to S-100. This protein was found in the spiral ligament, in Reissner's membrane, in the spiral limbus, and in the basement membrane of the cochlea. S-100-positive, thin fibers could be seen as supporting elements in the acoustic nerve and the facial nerve. This protein was localized along the surface of vestibular wall. The new technique provides an immunohistochemical approach to studying routinely processed human temporal bone sections and may prove useful in the field of immuno-otopathology.
- Published
- 1992
34. A technique for retrieving antigens in formalin-fixed, routinely acid-decalcified, celloidin-embedded human temporal bone sections for immunohistochemistry
- Author
-
C Coté, Krishan L. Kalra, A K Tandon, Shan-Rong Shi, and Clive R. Taylor
- Subjects
Pathology ,medicine.medical_specialty ,Tissue Fixation ,Histology ,medicine.drug_class ,Antibodies, Monoclonal ,Collodion ,Temporal Bone ,Hydrogen-Ion Concentration ,Monoclonal antibody ,Immunohistochemistry ,Stain ,chemistry.chemical_compound ,Antigen ,Antigen retrieval ,chemistry ,Formaldehyde ,Temporal bone ,Monoclonal ,medicine ,Humans ,Indicators and Reagents ,Anatomy - Abstract
The application of immunohistochemistry to routinely decalcified, celloidin-embedded human temporal bone sections has been hampered because of antigen loss during processing of the specimens. To our knowledge, there has been no published report to date describing immunohistochemical staining of such tissues suitable for examination by light microscopy. Here we report a novel antigen retrieval technique which can be successfully used to stain a variety of antigens in routinely formalin-fixed, trichloroacetic acid-decalcified, celloidin-embedded human temporal bone sections. The new procedure reported here for decalcified human temporal bone tissues simply requires immersing slides for 30 min at room temperature in an antigen retrieval solution. A total of 60 decalcified, celloidin-embedded human temporal bone tissues were tested with monoclonal antibodies (MAb) to 15 different antigens. Of these, 12 MAb showed definite positive staining, while three were negative. This technique may prove very useful in studying the expression of various antigens by immunohistochemistry in formalin-fixed, acid-decalcified, celloidin-embedded tissues.
- Published
- 1992
35. Immunohistochemistry
- Author
-
Debra Hawes, Richard J. Cote, Shan-Rong Shi, David J. Dabbs, and Clive R. Taylor
- Subjects
Pathology ,medicine.medical_specialty ,business.industry ,Immunohistochemistry ,Medicine ,business - Published
- 2009
36. Contributors
- Author
-
Charles A. Amezcua, Mahul B. Amin, Daniel A. Arber, Sylvia L. Asa, James B. Atkinson, Paul L. Auclair, Michael J. Becich, David G. Bostwick, Thomas W. Bouldin, Allen Burke, R. Tucker Burks, Norman J. Carr, John K.C. Chan, Karen L. Chang, Liang Cheng, Richard J. Cote, Antonio L. Cubilla, David J. Dabbs, Stephen J. DeArmond, John N. Eble, Gary L. Ellis, Robert A. Erlandson, Juan C. Felix, Wendy L. Frankel, Noriyoshi Fukushima, David A. Gaskin, John R. Gilbertson, William C. Gross, Farnaz Hasteh, Debra Hawes, David R. Hinton, Ralph H. Hruban, Mahlon D. Johnson, Cynthia G. Kaplan, Michael N. Koss, Michael Kyriakos, Sean K. Lau, David Lewin, Klaus J. Lewin, Grace Lin, Kurt Matthews, Isabelle Meiers, Martin C. Mihm, Anirban P. Mitra, Cesar A. Moran, Christopher A. Moskaluk, Lucien E. Nochomovitz, David A. Owen, Anil V. Parwani, Zdena Pavlova, Michael Peterson, Robert E. Petras, José Antonio Plaza, Victor G. Prieto, Mahendra Ranchod, Narsing A. Rao, Joseph A. Regezi, Mary Richardson, Robert R. Rickert, William B. Ross, Sharda G. Sabnis, Eric Schubert, Shan-Rong Shi, Jeffrey P. Simko, Leslie H. Sobin, Somsiri Sukavatcharin, Saul Suster, Pheroze Tamboli, Clive R. Taylor, Lester D.R. Thompson, Satish K. Tickoo, Thomas A. Tousseyn, David B. Troxel, Loretta L.Y. Tse, Renu Virmani, M. Kay Washington, Noel Weidner, Lawrence M. Weiss, Bruce M. Wenig, William O. Whetsell, Sharon P. Wilczynski, Robb E. Wilentz, Tai-Yuen Wong, and Thomas C. Wright
- Published
- 2009
37. [Current perspectives on immunohistochemistry]
- Author
-
Shan-rong, Shi and Bing-quan, Wu
- Subjects
Internationality ,Evaluation Studies as Topic ,Academies and Institutes ,Humans ,Immunohistochemistry ,Carcinoma in Situ - Published
- 2008
38. Antigen retrieval for proteomic characterization of formalin-fixed and paraffin-embedded tissues
- Author
-
Cheng S. Lee, Ying Liu, Xueping Fang, Li Yang, Clive R. Taylor, Shan-Rong Shi, Cheng Liu, Weijie Wang, Haifeng Xu, and Brian M. Balgley
- Subjects
Proteomics ,Paraffin Embedding ,Cluster of differentiation ,Isoelectric focusing ,Electrophoresis, Capillary ,Shotgun ,General Chemistry ,Computational biology ,Biology ,Biochemistry ,Molecular biology ,chemistry.chemical_compound ,Antigen retrieval ,chemistry ,Formaldehyde ,Proteome ,Immunohistochemistry ,Humans ,Antigens ,Shotgun proteomics - Abstract
Formalin-fixed and paraffin-embedded tissues represent the vast majority of archived tissue. Access to such tissue specimens via shotgun-based proteomic analyses may open new avenues for both prospective and retrospective translational research. In this study, we evaluate the effects of fixation time on antigen retrieval for the purposes of shotgun proteomics. For the first time, we demonstrate the capability of a capillary isotachophoresis (CITP)-based proteomic platform for the shotgun proteomic analysis of proteins recovered from FFPE tissues. In comparison to our previous studies utilizing capillary isoelectric focusing, the CITP-based analysis is more robust and increases proteome coverage. In this case, results from three FFPE liver tissues yield a total of 4098 distinct Swiss-Prot identifications at a 1% false-discovery rate. To judge the accuracy of these assignments, immunohistochemistry is performed on a panel of 17 commonly assayed proteins. These proteins span a wide range of protein abundances as inferred from relative quantitation via spectral counting. Among the panel were 4 proteins identified by a single peptide hit, including three clusters of differentiation (CD) markers: CD74, CD117, and CD45. Because single peptide hits are often regarded with skepticism, it is notable that all proteins tested by IHC stained positive.
- Published
- 2008
39. Antigen Retrieval Immunohistochemistry Based Research and Diagnostics
- Author
-
Shan-Rong Shi, Clive R. Taylor, Shan-Rong Shi, and Clive R. Taylor
- Subjects
- Antigens, Immunohistochemistry
- Abstract
The most complete, up-to-date reference on antigen retrieval and immunohistochemistry An antigen is a substance that prompts the generation of antibodies and can cause an immune response. The antigen retrieval (AR) technique is in wide use across the globe, and is a critical technique used in medical diagnosis of disease, particularly clinical targeted cancer treatment. Antigen Retrieval Immunohistochemistry Based Research and Diagnostics discusses several scientific approaches to the standardization of quantifiable immunohistochemistry (IHC). Based on the development and application of AR by the editors, this volume summarizes recent achievements in AR-IHC and analyzes numerous cutting-edge issues for future research projects. Featuring contributions from a worldwide group of leading experts and research scientists in the field, this important work: Summarizes the key problems in the four fields of antigen retrieval Discusses the advances of AR techniques and their applications Provides practical methods and protocols in AR-IHC, such as extraction of nucleic acids and proteins for molecular analysis, cell/tissue sample preparation, and standardization and development of various techniques to meet the future needs of¿clinical and research molecular analysis Encourages further research in AR and IHC, particularly how AR methods might be employed for improved test performance and the development of greater reliability and reproducibility of IHC Includes an appendix of related laboratory protocols Antigen Retrieval Immunohistochemistry Based Research and Diagnostics is intended for clinical pathologists, molecular cell biologists, basic research scientists, technicians, and graduate students who undertake tissue/cell morphologic and molecular analysis and wish to use and extend the power of immunohistochemistry. It is also pertinent for most biotechnology companies majoring in development of IHC products. Wiley Series in Biomedical Engineering and Multi-Disciplinary Integrated Systems / Kai Chang, Series Editor
- Published
- 2010
40. Immuno-Electron Microscopic Study of Keratin Distribution in the Cochlea Using Monoclonal Antibody
- Author
-
Iwao Ohtani, Michio Kobari, Shan-Rong Shi, and Tohru Aikawa
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Spiral limbus ,Guinea Pigs ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Keratin ,Border cells ,otorhinolaryngologic diseases ,medicine ,Animals ,Inner ear ,Microscopy, Immunoelectron ,030223 otorhinolaryngology ,Cochlea ,chemistry.chemical_classification ,integumentary system ,030102 biochemistry & molecular biology ,Antibodies, Monoclonal ,General Medicine ,Immunohistochemistry ,Cell biology ,medicine.anatomical_structure ,Otorhinolaryngology ,chemistry ,Cytoplasm ,Reticular connective tissue ,Ultrastructure ,Keratins ,sense organs - Abstract
Keratin distribution in the cochlea has been studied immuno–electron microscopically by both pre-embedding and postembedding methods. Keratin immunoreactivity in the guinea pig cochlea was clearly demonstrated in Hensen's cells, the reticular lamina surrounding both outer and inner hair cells, outer and inner pillar cells, Claudius' cells, inner and external sulcus cells, interdental cells of the spiral limbus, Reissner's membrane, border cells, inner phalangeal cells, Deiters' cells, and spiral prominence cells. Keratin expression at the ultrastructural level showed a nonfilamentous keratin system in the cytoplasm of cochlear supporting cells.
- Published
- 1990
41. Development of an optimal antigen retrieval protocol for immunohistochemistry of retinoblastoma protein (pRB) in formalin fixed, paraffin sections based on comparison of different methods
- Author
-
Cr Taylor, Shan Rong Shi, C Liu, and Lillian L. Young
- Subjects
Pathology ,medicine.medical_specialty ,Histology ,Retinoblastoma Protein ,Sensitivity and Specificity ,chemistry.chemical_compound ,Antigen ,Cell Line, Tumor ,Formaldehyde ,Paraffin section ,medicine ,Humans ,Antigens ,Paraffin Embedding ,biology ,Staining and Labeling ,Chemistry ,Retinoblastoma protein ,Reproducibility of Results ,General Medicine ,Formalin fixed ,Molecular biology ,Staining ,Medical Laboratory Technology ,Tissue sections ,Antigen retrieval ,Urinary Bladder Neoplasms ,biology.protein ,Immunohistochemistry - Abstract
A novel protocol for antigen retrieval (AR) for immunohistochemistry (IHC) of retinoblastoma protein (pRB) in formalin fixed, paraffin embedded (FFPE) tissue sections was developed using 0.05% citraconic anhydride as the AR solution for heat treatment based on comparison of different methods. This new protocol has advantages including superior morphological preservation, greater reproducibility, and more intense staining after retrieval. Our study demonstrates the importance of comparing various AR protocols to obtain maximal IHC for standardization and for quantitative IHC.
- Published
- 2007
42. [Updates on antigen retrieval techniques for immunohistochemistry]
- Author
-
Shan-rong, Shi, Yan, Shi, and Clive R, Taylor
- Subjects
Fixatives ,Hot Temperature ,Paraffin Embedding ,Tissue Fixation ,Staining and Labeling ,Formaldehyde ,Humans ,Proteins ,Antigens ,Immunohistochemistry ,In Situ Hybridization - Published
- 2007
43. Evaluation of the Value of Frozen Tissue Section Used as 'Golden Standard' for Immunohistochemistry
- Author
-
Cheng Liu, Laurie Yee Mae Tang, Ryan Chen, Shan-Rong Shi, Richard J. Cote, Andrew S. Young, Llana Pootrakul, and Clive R. Taylor
- Subjects
Section (archaeology) ,business.industry ,Genetics ,Immunohistochemistry ,Frozen tissue ,Nuclear medicine ,business ,Molecular Biology ,Biochemistry ,Value (mathematics) ,Biotechnology ,Mathematics - Published
- 2007
44. Expression of stress response protein Grp78 is associated with the development of castration-resistant prostate cancer
- Author
-
Llana Pootrakul, Ram H. Datar, Jie Cai, Amy S. Lee, Debra Hawes, Shan Rong Shi, Susan Groshen, and Richard J. Cote
- Subjects
PCA3 ,Male ,Cancer Research ,medicine.medical_specialty ,Cohort Studies ,Prostate cancer ,Castration Resistance ,Prostate ,Stress, Physiological ,Internal medicine ,LNCaP ,Tumor Cells, Cultured ,Medicine ,Humans ,Orchiectomy ,Endoplasmic Reticulum Chaperone BiP ,Heat-Shock Proteins ,Neoplasm Staging ,Tumor microenvironment ,business.industry ,Prostatic Neoplasms ,medicine.disease ,Gene Expression Regulation, Neoplastic ,Endocrinology ,medicine.anatomical_structure ,Oncology ,Cancer cell ,Cancer research ,business ,Molecular Chaperones - Abstract
Background: Induction of molecular chaperone Grp78 (78-kDa glucose-regulated protein) occurs in stress conditions that often characterize tumor microenvironments. We investigated the role of Grp78 in prostate cancer progression and the development of castration resistance, where cancer cells continue to survive despite the stress of an androgen-starved environment. Experimental Design: Immunohistochemistry was done to examine Grp78 expression in 219 prostate cancers from patients with pathologic stage T3N0M0 disease [androgen ablation naive (untreated) and androgen ablation exposed (treated)] and castration-resistant prostate cancer. Classification of tumors was based on intensity of Grp78 cytoplasmic immunoreactivity and percentage of immunoreactive tumor cells. The associations of Grp78 expression with prostate cancer recurrence (clinical and/or serum prostate-specific antigen) and survival were examined in the untreated stage T3N0M0 group. Grp78 expression was also analyzed in the androgen-dependent LNCaP and castration-resistant C42B cell lines. Results: The percentage of tumor cells expressing Grp78 was strongly associated with castration-resistant status (P = 0.005). Increased Grp78 expression was consistently associated with greater risk of prostate cancer recurrence and worse overall survival in patients who had not undergone prior hormonal manipulation. Grp78 expression was also increased in the castration-resistant LNCaP-derived cell line C42B and in LNCaP cells grown in androgen-deprived conditions compared with LNCaP cells grown in androgen-rich media. Conclusion: Our findings show that up-regulation of Grp78 is associated with the development of castration resistance, possibly in part by augmenting cell survival as previously suggested, and may serve as an important prognostic indicator of recurrence in a subset of patients with T3N0M0 disease.
- Published
- 2006
45. Standardization of immunohistochemistry for formalin-fixed, paraffin-embedded tissue sections based on the antigen-retrieval technique: from experiments to hypothesis
- Author
-
Clive R. Taylor, Shan-Rong Shi, and Cheng Liu
- Subjects
Pathology ,medicine.medical_specialty ,Histology ,Time Factors ,Formalin fixed paraffin embedded ,Biology ,chemistry.chemical_compound ,Fixatives ,Fresh Tissue ,Formaldehyde ,medicine ,Humans ,Paraffin embedding ,Antigens ,Fixation (histology) ,Paraffin Embedding ,Proteins ,Reproducibility of Results ,Molecular biology ,Immunohistochemistry ,Staining ,Tissue sections ,Antigen retrieval ,chemistry ,Anatomy - Abstract
From a practical point of view, one of the most difficult issues in the standardization of IHC for FFPE tissue is the adverse influence of formalin upon antigenicity, as well as the great variation in fixation/processing procedures. Based on previous study, an additional study using four markers demonstrated the potential for obtaining equivalent IHC staining among FFPE tissue sections with periods of formalin fixation ranging from 6 hr to 30 days. On this basis, the following hypothesis is proposed. "The use of optimized AR protocols permits retrieval of specific proteins (antigens) from FFPE tissues to a defined and reproducible degree (expressed as R%), with reference to the amount of protein present in the original fresh/unfixed tissue". This hypothesis may also be presented mathematically: the protein amount in a fresh cell/tissue, expressed as Pf, produces an IHC signal in fresh tissue of integral(Pf). When the identical IHC staining plus AR treatment is applied to a FFPE tissue section, the IHC signal may be represented as integral (Pffpe). The degree of retrieval after AR (R%) is calculated as follows: R% = integral (Pffpe)/ integral (Pf) x 100%. The amount of protein in the FFPE tissue may then be derived as follows: Pffpe = Pf x R%. In a situation where optimized AR is 100% effective, the IHC signal would then be of equal strength in fresh tissue and FFPE tissue, and Pffpe= Pf. Further studies are designed to test the limitations of the proposed hypothesis.
- Published
- 2006
46. Contributors
- Author
-
Leon Barnes, Nancy J Barr, Deborah Belchis, Parul Bhargava, David S Bosler, David G Bostwick, Lisa A Cerilli, Cheryl Coffin, David J Dabbs, Ronald A DeLellis, Eduardo J Eyzaguirre, Christopher Gocke, Neal S Goldstein, Samuel P Hammar, Jennifer L Hunt, Christina Isacson, Deborah Josefson, Marshall E Kadin, Lina Liu, Jun Ma, Paul E McKeever, James W Patterson, Junqi Qian, Shan-Rong Shi, Sandra J Shin, Robert A Soslow, Paul E Swanson, Clive R Taylor, David Walker, Mark R Wick, Nancy Wu, Sherif R Zaki, and Charles F Zaloudek
- Published
- 2006
47. Techniques of Immunohistochemistry: Principles, Pitfalls and Standardization
- Author
-
Clive R. Taylor, Nancy J. Barr, Nancy Wu, and Shan-Rong Shi
- Subjects
medicine.medical_specialty ,Standardization ,Chemistry ,medicine ,Immunohistochemistry ,Medical physics - Published
- 2006
48. DNA extraction from archival formalin-fixed, paraffin-embedded tissues: heat-induced retrieval in alkaline solution
- Author
-
Ram H. Datar, Shan Rong Shi, Lin Wu, Zina Zhang, Richard J. Cote, Cheng Liu, and Clive R. Taylor
- Subjects
Histology ,Tissue Fixation ,Polymerase Chain Reaction ,Buffer (optical fiber) ,Heating ,chemistry.chemical_compound ,Fixatives ,Spectrophotometry ,Boiling ,Formaldehyde ,medicine ,Humans ,Antigens ,Molecular Biology ,Chromatography ,Paraffin Embedding ,medicine.diagnostic_test ,Cell Biology ,Buffer solution ,DNA ,Hydrogen-Ion Concentration ,DNA extraction ,Medical Laboratory Technology ,Electrophoresis ,chemistry ,Antigen retrieval - Abstract
Based on the antigen retrieval principle, our previous study has demonstrated that heating archival formalin-fixed, paraffin-embedded (FFPE) tissues at a higher temperature and at higher pH value of the retrieval solution may achieve higher efficiency of extracted DNA, when compared to the traditional enzyme digestion method. Along this line of heat-induced retrieval, this further study is focused on development of a simpler and more effective heat-induced DNA retrieval technique by testing various retrieval solutions. Three major experiments using a high temperature heating method to extract DNA from FFPE human lymphoid and other tissue sections were performed to compare: (1) different concentrations of alkaline solution (NaOH or KOH, pH 11.5–12) versus Britton and Robinson type of buffer solution (BR buffer) of pH 12 that was the only retrieval solution tested in our previous study; (2) several chemical solutions (SDS, Tween 20, and GITC of various concentrations) versus BR buffer or alkaline solution; and (3) alkaline solution mixed with chemicals versus BR buffer or single alkaline solution. Efficiency of DNA extraction was evaluated by measuring yields using spectrophotometry, electrophoretic pattern, semiquantitation of tissue dissolution, PCR amplification, and kinetic thermocycling-PCR methods. Results showed that boiling tissue sections in 0.1 M NaOH or KOH or its complex retrieval solutions produced higher yields and better quality of DNA compared to BR buffer or chemical solutions alone. The conclusion was that boiling FFPE tissue sections in 0.1 M alkaline solution is a simpler and more effective heat-induced retrieval protocol for DNA extraction. Combination with some chemicals (detergents) may further significantly improve efficiency of the heat-induced retrieval technique.
- Published
- 2004
49. Antigen Retrieval Technique for Immunohistochemistry: Principles, Protocols, and Further Development
- Author
-
Shan-Rong Shi and Clive R. Taylor
- Subjects
Pathology ,medicine.medical_specialty ,chemistry.chemical_compound ,Antigen retrieval ,chemistry ,medicine ,Immunohistochemistry - Published
- 2003
50. Use of Monoclonal Antibodies in Immunohistochemistry
- Author
-
Atul K. Tandon, Krishan L. Kalra, Jeffrey B. Prince, Christopher Jones, and Shan-Rong Shi
- Subjects
Pathology ,medicine.medical_specialty ,business.industry ,medicine.drug_class ,Medicine ,Immunohistochemistry ,business ,Monoclonal antibody - Published
- 2003
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