Journal of Pathology and Translational Medicine

Original Article

Multistaining Optimization for Epstein-Barr Virus–Encoded RNA In Situ Hybridization and Immunohistochemistry of Formalin-Fixed Paraffin-Embedded Tissues Using an Automated Immunostainer: Jae Nam Ko, Jin Kyoung Jung, Yun Ik Park, Hwa Jeong Shin, Jooryung Huh, Sol Back, Yu Jin Kim, Jae Ho Kim, Heounjeong Go; J Pathol Transl Med. 2019;53(5):317-326. Published online August 27, 2019; DOI: https://doi.org/10.4132/jptm.2019.08.06

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Background
Single staining is commonly performed for practical pathologic diagnoses. However, this method is limited in its ability to specify cellular morphology and immunophenotype and often requires consumption of limited tissue. This study aimed to describe an optimized protocol for multiple in situ hybridization (ISH) and immunohistochemistry (IHC).
Methods
The quality of multistaining was evaluated by carefully changing each step of ISH and IHC in an angioimmunoblastic T-cell lymphoma (AITL) case on a Ventana BenchMark XT automated immunostainer. The optimized protocols were also performed using another immunostainer and in 15 cases of five Epstein-Barr virus (EBV)–associated malignancies using formalin-fixed paraffin-embedded tissue.
Results
The quality of various ISHIHC staining protocols was semi-quantitatively evaluated. The best EBV-encoded RNA (EBER)-ISH/double IHC staining quality, equivalent to single staining, was obtained using the following considerations: initial EBER-ISH application, use of protease and antigen retrieval reagent (cell conditioning 1 [CC1] treatment time was minimized due to impact on tissue quality), additional baking/ deparaffinization not needed, and reduced dilution ratio and increased reaction time for primary antibody compared with single immunostaining. Furthermore, shorter second CC1 treatment time yielded better results. Multiple staining was the best quality in another immunostainer and for different types of EBV-associated malignancies when it was performed in the same manner as for the Ventana BenchMark XT as determined for AITL.
Conclusions
EBER-ISH and double IHC could be easily used in clinical practice with currently available automated immunostainers and adjustment of reagent treatment time, dilution ratio, and antibody reaction time.

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Shilah A. Bonnett, Alyssa B. Rosenbloom, Giang T. Ong, Mark Conner, Aric B.E. Rininger, Daniel Newhouse, Felicia New, Chi Q. Phan, Saskia Ilcisin, Hiromi Sato, John S. Lyssand, Gary Geiss, Joseph M. Beechem
Cancer Research Communications.2023; 3(5): 763. CrossRef
Detection of Epstein–Barr Virus in Periodontitis: A Review of Methodological Approaches
Lilit Tonoyan, Marlène Chevalier, Séverine Vincent-Bugnas, Robert Marsault, Alain Doglio
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Review

Let Archived Paraffin Blocks Be Utilized for Research with Waiver of Informed Consent: Yong-Jin Kim, Jeong Sik Park, Karam Ko, Chang Rok Jeong; J Pathol Transl Med. 2018;52(3):141-147. Published online April 5, 2018; DOI: https://doi.org/10.4132/jptm.2018.02.07

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Abstract PDF

Advances in biomedical and genetic research have contributed to more effective public health improvement via bench-to-bed research and the emergence of personalized medicine. This has certainly showcased the importance of archived human tissues, especially paraffin-embedded blocks in pathology. Currently in Korea, undue legislative regulations of the Bioethics and Safety Act suspend and at times discourage studies from taking place. In this paper, the authors underline the value of paraffin blocks in the era of personalized and translational medicine. We discuss detailed clauses regarding the applicability of paraffin blocks from a legal perspective and compare Korea’s regulations with those of other countries. The necessity for allowing waived consent and Institutional Review Board (IRB) approval will be argued throughout. The authors suggest that researchers declare the following to obtain IRB approval and waiver of informed consents: research could not be practically carried out without a waiver of consent; the proposed research presents no more than minimal risk of harm to subjects, and the waiver of consent will not adversely affect the rights and welfare of subjects; and research will not utilize a tissue block if only 1 is available for each subject, to allow future clinical use such as re-evaluation or further studies.

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NaV1.7 channels are expressed in the lower airways of the human respiratory tract
Everardo Hernández-Plata, Ana Alfaro Cruz, Carina Becerril
Respiratory Physiology & Neurobiology.2023; 311: 104034. CrossRef
Expression Profiles of GILZ and SGK-1 in Potentially Malignant and Malignant Human Oral Lesions
Mahmood S. Mozaffari, Rafik Abdelsayed
Frontiers in Oral Health.2021;[Epub] CrossRef
IRB review points for studies utilizing paraffin blocks archived in the pathology laboratory
Yong-Jin Kim, Chang Rok Jeong, Jeong Sik Park
Yeungnam University Journal of Medicine.2018; 35(1): 36. CrossRef

Original Articles

In-house Manual Construction of High-Density and High-Quality Tissue Microarrays by Using Homemade Recipient Agarose-Paraffin Blocks: Kyu Ho Kim, Suk Jin Choi, Yeon Il Choi, Lucia Kim, In Suh Park, Jee Young Han, Joon Mee Kim, Young Chae Chu; Korean J Pathol. 2013;47(3):238-244. Published online June 25, 2013; DOI: https://doi.org/10.4132/KoreanJPathol.2013.47.3.238

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Abstract PDF

Background

Self-made tissue punches can be effectively used to punch holes in blank recipient paraffin blocks and extract tissue cores from the donor paraffin blocks for the low-cost construction of tissue microarrays (TMAs). However, variable degrees of section distortion and loss of the tissue cores can occurs during cutting of the TMAs, posing technical problems for in-house manual construction of high-density TMAs. We aimed to update the method for in-house manual TMA construction to improve the quality of high-density TMAs.

Methods

Blocks of agarose gel were subjected to the standard tissue processing and embedding procedure to prepare recipient agarose-paraffin blocks. The self-made tissue punches and recipient agarose-paraffin blocks were used to construct TMAs, which were completely melted and re-embedded in paraffin to make finished TMA blocks.

Results

The donor tissue cores were completely integrated into the surrounding paraffin of the recipient blocks. This method enabled us to construct high-density TMAs with significantly less section distortion or loss of tissue cores during microtomy.

Conclusions

Simple and inexpensive construction of high-density and high-quality TMAs can be warranted by using paraffinized agarose gels as recipient blocks.

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Optimization of Tissue Microarrays from Banked Human Formalin-Fixed Paraffin Embedded Tissues in the Cancer Research Setting
Tammy Sexton, Gregory L. Kucera, Edward A. Levine, Kounosuke Watabe, Stacey S. O'Neill
Biopreservation and Biobanking.2019; 17(5): 452. CrossRef
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Yan-Wei Cao, Yong Liu, Zhen Dong, Lei Guo, En-Hao Kang, Yong-Hua Wang, Wei Zhang, Hai-Tao Niu
Urologic Oncology: Seminars and Original Investigations.2018; 36(6): 311.e15. CrossRef
Platelet-derived growth factor receptor α in hepatocellular carcinoma is a prognostic marker independent of underlying liver cirrhosis
Jung-Hwan Yu, Joon Mee Kim, Ja Kyung Kim, Suk Jin Choi, Kwan Sik Lee, Jin-Woo Lee, Hye Young Chang, Jung Il Lee
Oncotarget.2017; 8(24): 39534. CrossRef
Prognostic Implication of Semi-quantitative Immunohistochemical Assessment of CD20 Expression in Diffuse Large B-Cell Lymphoma
Chang Hwan Choi, Young Hoon Park, Joo Han Lim, Suk Jin Choi, Lucia Kim, In Suh Park, Jee Young Han, Joon Mee Kim, Young Chae Chu
Journal of Pathology and Translational Medicine.2016; 50(2): 96. CrossRef
High Quality Tissue Miniarray Technique Using a Conventional TV/Radio Telescopic Antenna
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Construction of High-Density Tissue Microarrays at Low Cost by Using Self-Made Manual Microarray Kits and Recipient Paraffin Blocks: Chang Hwan Choi, Kyu Ho Kim, Ju Young Song, Suk Jin Choi, Lucia Kim, In Suh Park, Jee Young Han, Joon Mee Kim, Young Chae Chu; Korean J Pathol. 2012;46(6):562-568. Published online December 26, 2012; DOI: https://doi.org/10.4132/KoreanJPathol.2012.46.6.562

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Abstract PDF

Background

Advances of tissue microarray (TMA) technology have enabled simultaneous in situ analysis of biomarker expression in a large number of archived pathology specimens. However, the relatively high cost of TMA construction may hamper many researchers from using this essential tool of modern pathology research. We discuss methods for making TMA kits and recipient blocks for manual construction of high-density TMAs at low cost.

Methods

Ordinary cannula piercing needles, hypodermic needles, bone marrow biopsy needles, metallic ink cartridges of ballpoint pens, and disposable skin biopsy punches were used to construct self-made manual TMA kits. The recipient blocks were manufactured by boring holes in the conventional bare paraffin blocks. A mini electric hand drill and a microcompound table assembled on a drill stand were used to maximize the capacity of the recipient blocks.

Results

By using TMA kits made from cannula piercing needles (16- and 18-gauge), it was possible to construct TMAs with 1 mm×140 cores, 0.6 mm×320 cores, 2 mm×70 cores, 3 mm×35 cores, and 5 mm×12 cores. The capacity of the recipient blocks could be dramatically increased by drilling holes.

Conclusions

Construction of TMAs using self-made TMA kits is an inexpensive alternative to construction of TMAs using commercial devices.

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Biopreservation and Biobanking.2019; 17(5): 452. CrossRef
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Alessandra Sfacteria, Laura Perillo, Francesco Macrì, Giovanni Lanteri, Claudia Rifici, Giuseppe Mazzullo
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Hua Chang, Diane Peluso, Sadiq Hussain, Michail Shipitsin, Peter Blume-Jensen
Journal of Histotechnology.2014; 37(3): 95. CrossRef
In-house Manual Construction of High-Density and High-Quality Tissue Microarrays by Using Homemade Recipient Agarose-Paraffin Blocks
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Korean Journal of Pathology.2013; 47(3): 238. CrossRef

Clinical Value of Demonstration of Estrogen and Progesterone Receptors using Paraffin Wax Sections in Breast Carcinoma.: Hye Kyung Ahn, Yun Jung Kim, Young Euy Park; Korean J Pathol. 1996;30(2):89-93.

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Abstract PDF: This study aimed at assessing the usefulness of paraffin wax sections for demonstration of ER(estrogen receptor) and PR(progesterone receptor), using brief microwave processing rather than proteolytic predigestion. The receptor status of a breast cancer is often into consideration when planning treatment nowadays. As biochemical receptor assays require large amount of fresh tumor tissue and are not always available for all tumors, pathologists are now increasingly asked to provide a service for the assessment of the receptor status in tissue routine sections. Until recently, immunohistochemical demonstration of receptors was used in frozen sections. Therefore, routinely processed paraffin wax sections of 25 cases of breast carcinoma with known ER and PR concentrations, estimated by the standard DCC(dextran-coated charcoal) biochemical assay, were examined using the ABC immunoperoxidase technique. The results were assessed semiquantitably, using a five grade scoring system. Of the 25 cases examined, with DCC cutoff point being <10 fmol,71% and 75% in positivity of each ER, PR receptor is concordant. Statistic analysis demonstrates high relationship between scoring system of IH method and DCC value in ER (R=0.6061, p=0.001) and PR (R=0.5832, p=0.001). The IH method can provide easily assessed reliable positive information about ER, PR status of breast carcinoma using routinely processed paraffin wax sections.

Histopathological Differences between Silicone Granuloma and Paraffinoma.: Yeon Mee Kim, Hye Kyung Lee, Hye Je Cho, Je Geun Cho; Korean J Pathol. 1996;30(5):427-436.

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Abstract PDF: During the past two decades, silicone (polydimethylsiloxane) has become one of the most extensively applied biomaterials. Although pure silicone is relatively inert and usually causes only minimal tissue reactions, it has been reported to evoke a definite foreign body reaction. We studied five cases of silicone-induced granulomas in various sites; two in the breast, one in the breast and axillary lymph nodes, one in the subcutis of the abdomen, back and extremities and one in the eyeball, to illustrate the salient histopathologic features of reactions to silicone with particular emphasis to its differences from paraffin granuloma. For this, 17 paraffinomas were also studied. Tissue reaction to silicone liquid and gel was characterized by numerous round to oval empty cystic vacuoles, mild to moderate fat necrosis, foreign body reaction, a variable degree of mononuclear inflammatory cell infiltration and mild focal fibrosis. The cystic spaces were relatively uniform and showed a snow-man like appearance. In contrast to the silicone granulomas, the paraffinomas, also refered to as sclerosing lipogranulomas showed diffuse sclerosis and frequent calcification around the cystic vacuoles. The cystic spaces in paraffinomas were swiss cheese-like configuration, and the content of the cystic spaces was dirty and frequently calcified. However, there were certain similarities between these two types of granulomas particularly in the early phases of the reaction, therefore, the history of silicone injection or implant, is sometimes critical to the diagnosis of silicone granuloma. Despite great technologic advances in the manufacturing of prostheses and medical equipment, droplets and/or particles of silicone still escape into the body tissues in a variety of ways; therefores, the pathologist should always wonder whether the histologic reaction observed is due to silicone or to some other foreign material including paraffin.

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