Histotalks: Nsh Podcasts

Audio reading from the July 2023 NSH Fixation on Histology Blog, Hot Dog as an Alternative Source for Control Tissue. Read entire article.

P08-Diagnostic Cytopathology Cell Block Preparation Methods- Anna Patterson FIBMS MSc CSci Diagnostic Cytology Scheme Coordinator Helen Naylor MSc Diagnostic Cytology Technical Specialist Cell blocks from Cytopathology samples have always had value in the diagnostic process as a complement to the traditional Cytopathology stains – Papanicolaou and Romanowsky.  This has become more important to provide material for Immunocytochemistry to refine malignant diagnosis, and more recently, for the use of molecular testing to aid in the choice of tailored chemotherapy regimens.  If this information can be obtained from Cytopathology samples, which are less invasive than biopsy samples, the patient will benefit. A variety of preparation methods are available for the preparation of cell blocks from cells from Diagnostic Cytopathology samples.  The most popular methods will be discussed and how they can be used to optimise the quality of cell preservation if used correctly. Information garnered from the results of th

P06-Understanding the Quality of your Electron Microscopy Provider in this Era of Outsourcing of Services- Tracey de Haro MSc, FIBMS, UK NEQAS CPT TEM Scheme Coordinator Specialist Scientific Lead for Electron Microscopy University Hospitals Of Leicester NHS Trust, UK  Background Electron Microscopy (EM) remains vital to the diagnostic repertoire for the diagnosis of pathologies. Surveys of diagnostic TEM units in the UK were carried out in 2012, in 2019 (unpublished) and is currently being repeated. These surveys showed that a large amount of EM is now outsourced to units away from the originating trust. Whether UK or globally, when pathology departments are looking for a supplier of diagnostic EM services, the only questions they ask of the EM units are “how much does it cost” and “what is your turnaround time?” Are these the only relevant questions to ask? Considerations The following relevant issues should be considered alongside cost and speed; The technical quality of an electron microscopy service;

The Development of a Cocktail of Microglia and GFAP For Easy Diagnosis - Anisha Bhasin B.S, Sarah Holguin, MBA. B.S, Joe Vargas, M.S Microglia and GFAP are distinct neural markers, typically used separately to diagnose the degree of neurological infection and injury. Microglia, a glial cell, is used in the immune response of the central nervous system. GFAP is an astrocyte marker; astrocytes provide structural support and make up the blood-brain barrier. Using the two in conjugation with one another would prove to be an efficient diagnostic tool. A cocktail was constructed with optimal titration to observe the two markers in unison. In clinical usage, it will provide an efficient diagnosis of chronic inflammatory conditions of the central nervous system. The staining was conducted in IHC and fluorescence to compare morphology and count. Due to anatomical similarities, there tends to be morphological confusion between microglia and GFAP. However, when stained in conjunction with one another, notable differe

Study tools for the histotechnologist (HTL) Histotechnician (HT) certification exam- Amber Moser1, BS, Hannah Benton1, BS, Taylor Wallace1, BS, and Elizabeth A. Chlipala1, BS, HTL(ASCP)QIHC Premier Labs, Longmont, CO Histology laboratories have seen an increase in workplace shortages since the COVID-19 pandemic and an increase in noncertified applicants to fill open positions. The traditional route for histology certification is the completion of a histology education series with an accredited institution; however, there are many other routes to qualify for certification. With declines in histology program graduate numbers, and the change in work force after the COVID-19 pandemic, it is expected that nontechnical positions will continue to be filled by noncertified individuals. This may lead to an increase in alternative routes of certification, specifically through laboratory experience. These tools provide additional resources for individuals studying for the HT/HTL certification exams. They are cate

Somagen Method of Alcohol Reagent Tracking (SMART) Optimization to Realize Workflow Efficiencies within the Laboratory-Anne de Wolf, BSc MLT Somagen Diagnostics, Edmonton, Alberta, Canada; Christine Chiu MLT Somagen Diagnostics, Toronto, Ontario, Canada; Annie Vallee Somagen Diagnostics, Montreal, Quebec, Canada; James Tang, BSc MLT Somagen Diagnostics, Edmonton, Alberta, Canada Tissue processing in recent decades has changed without subsequent changes to reagent maintenance. Reagent maintenance protocols have been ‘Grandfathered down’ without consideration for the reagent quality and how this is related to tissue types and block numbers processed. The concentration of the last absolute alcohol station needs to be >98 % to effectively remove water from tissue before clearing and paraffin infiltration.1, 2 Using the Eagle Eye Digital hydrometer can provide reagent quality information which could also provide potential downstream efficiencies to laboratory workflow and cost savings in labour and reagen

Method for FFPE Organ-Chip -Lindsay Parmelee HTL(ASCP) 1,2,3, Stephanie Pei Tung Yiu PhD 1,2,3,4, Chi Ngai Chan PhD 1,2,3, Sizun Jiang PhD 1,2,31 Center for Virology and Vaccine Research 2 Beth Israel Deaconess Medical Center 3 Harvard Medical School 4 Wyss Institute Organoids are currently being developed for applications in biomedicine such as drug development and disease research. With many organoid models emerging on the market there is growing need to develop methods to adapt organoids to the established work flows and assays that are the foundations of modern research.  A method was developed for processing, embedding, and sectioning Organ-Chips for FFPE workflows. Organ-Chips were treated with Histogel and trimmed to fit standard sized cassettes. Followed by processing and embedding as usual before proceeding to the microtome. The resulting slides have successfully been stained with hematoxylin and eosin and immunohistochemistry, as well as shown promising results for use on multiplexing platform

Assessing Adhesion Slide Performance Across Histology Applications -Colin Brewer, CellPath, Newtown, Wales; Rachel Finn, HTL, StatLab, McKinney, TX; Neil Haine, PhD, CellPath, Newtown, Wales; Arielle Hobson, StatLab, McKinney, TX; Moritz Kamphenkel, Knittel Glass, Braunschweig, Germany; Ronja-Melinda Komoll, Ph.D, Knittel Glass, Braunschweig, Germany; Racheal Moore, HT, StatLab, McKinney, TX; Nicole Romer, Knittel Glass, Braunschweig, Germany; Edeltraud Schikora, Knittel Glass, Braunschweig, Germany; Stefan Welsch, Knittel Glass, Braunschweig, Germany; Susan Willis, CellPath, Newtown, Wales Adhesion slides are widely preferred for IHC to aid in securing tissue sections to the slide and prevent reworks that could potentially postpone a patient diagnosis and drive-up costs in the lab. The cost of reworking a failed IHC slide due to poor tissue adhesion is estimated to be ~$80 per slide, considering the reagent cost and workload administration.1 Adhesion slides reinforce tissue adherence and integrity, minimi

The Use of Immunohistochemistry vs. qPCR to Detect BRAFV600E in Thyroid Cancer- Kannitha Chek HTL (ASCP), Nicholas Hoo-Fatt, MS, HTL (ASCP), DP, Samantha Diamond, M.D., Haresh Mani, M.D., Myong Ho “Lucy” Nam, M.D. Introduction: This study aimed to validate the effectiveness of the Ventana anti-BRAFV600E antibody in detecting the BRAFV600E mutation in thyroid cancer using immunohistochemistry (IHC) as an initial screening test. The BRAFV600E mutation is a common genetic alteration in thyroid cancer, particularly papillary thyroid carcinoma (PTC), and detection is crucial for prognosis and treatment decisions. Methods: The research examined 12 thyroid cancer cases (11 PTC and 1 anaplastic thyroid carcinoma) and 12 non-cancerous thyroid cases. This study was conducted by using both the IHC BRAFV600E antibody as well as genetic testing methods including Next-Generation Sequencing (NGS) and the Biocartis Idylla rapid qPCR module for BRAF mutation. Results: The antibody successfully identified the BRAFV600E

Whole NHP Brain Microtomy Technique- S Wessel, MS, HTL (ASCP),AskBio, Discovery 1, Morrisville, North Carolina T Marais, AskBio France, Paris Brain Institute (ICM), Paris, France M Martini, PhD, PhD, AskBio, Discovery 1, Morrisville, North Carolina, M Faridounnia, PhD, MSc, AskBio, Discovery 1, Morrisville, North Carolina, D Gregorowicz, HTL (ASCP),AskBio, Discovery 1, Morrisville, North Carolina, L Haertel, HT (ASCP), AskBio, Discovery 1, Morrisville, North Carolina M Hamilton, HTL (ASCP), AskBio, Discovery 1, Morrisville, North Carolina, M Vallucci, AskBio France, Paris Brain Institute (ICM), Paris, France ,S Alves, PhD, MSc, AskBio France, Paris Brain Institute (ICM), Paris, France, J Mysore, BVSc, MVSc, PhD, DACVP, AskBio, Discovery 1, Morrisville, North Carolina Preserving the integrity and conformation of capsid proteins, enzymes, and antigens while working with large tissue sections presents a “big” challenge in histology laboratories. With our Whole NHP (Non-Human Primate) Brain microtomy technique

Encapsulated Cell Therapies in Sheep as a Translational Model - Trisha M. Fabijanic, B.S., University of Arizona, Tucson, AZ, Amy C. Kelly, Ph.D, University of Arizona, Tucson, AZ, Eliza H. Johnson, University of Arizona, Tucson, AZ, Delaney A. Drew, B.S., University of Arizona, Tucson, AZ, Demetri A. Vlachos, B.S., University of Arizona, Tucson, AZ, Charles W. Putnam, M.D., Ph.D., University of Arizona, Tucson, AZ, Sean W. Limesand, Ph.D., University of Arizona, Tucson, AZ, Klearchos K. Papas, Ph.D., University of Arizona, Tucson, AZ Introduction: Cell therapy has become increasingly studied for therapeutic use in drug delivery and regenerative medicine. Devices intended for implantation of cells require construction of membranes that can safely contain transplanted cells, provide a barrier to the host immune system and an outer vascularizing membrane to induce close vessel formation. Sheep make an excellent model because of their nearly comparable size, physiology, and disease profile with humans, as

Cellular localization of the pink bollworm Cry1Ac Bt protein receptor PgCad1 in cultured insect cells Authors: Melanie Miranda (BS, HTL- ASCP, University of Arizona-Comparative Pathology Core Laboratory, Tucson, AZ) Jeff Fabrick (PhD, United States Department of Agriculture, Phoenix, AZ) Transgenic cotton genetically engineered to produce insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are used to manage insect pests, including the pink bollworm (Pectinophora gossypiella), a globally invasive pest of cotton. However, the evolution of resistance to Bt Cry proteins (Cry1Ac and Cry2Ab) by the pink bollworm in parts of Asia has reduced the effectiveness of Bt cotton. Several mutations in the midgut cadherin gene PgCad1 are genetically linked with the resistance of the Cry1Ac Bt toxin, which can result in disrupted cellular trafficking of the cadherin receptor to the surface of the midgut membrane and decrease toxin binding. Here, we established specialized insect histology and immunohistoche

2023 NSH co-presenters Gabriela De la Cruz and Lauren Ralph give great advice on selecting a topic and share why they decided to co-present at the 2023 NSH Convention. 

NSH member Ariel (Ari) Liberda discusses her decision to present at the 2023 NSH Convention.  She shares her journey and advice to anyone who is considering taking the leap!

Making the FNA Count Author: Jeffery Rinker The hospital at Sanford Bemidji Medical Center has 119 beds and performs an average of 150 FNAs each year. Our histologists do both histology and cytology, with FNAs comprising the majority of our cytology. Since 2020, our non-diagnostic rates on FNA samples have fluctuated. In 2022, our rate was as high as 28%. Compared to the non-diagnostic rate of 10% outlined in The Bethesda System for Reporting Thyroid Cytopathology, our results were over two times the accepted rate. To bring non-diagnostic rates down at our facility, a look at processing from beginning to end.  Radiology was the first place to start an overview of the process. During observation, we discovered that multiple passes of the needle contributed the bloodier samples, which affected the specimens processed later. To counter this, we reduced the number of smears and increased the amount of specimen deposited into CytoLyt (ThinPrep). Next, the cytology department process was examined. During this exami

Alternative Strategies For Analyzing Pre-Clinical Mouse Lungs Authors: Nicholas Pankow, BA; Gabriela De la Cruz, BS; Hannah Marie Atkins, PhD, DVM In some diseases or conditions, it is challenging to leverage human tissues to determine patterns and resolve disease progression. Similarities between human and mouse biology make using murine models in pre-clinical studies possible. More specifically, mouse lungs are used to further analyze respiratory illnesses as they provide scalable models that can either be genetically manipulated to elicit human diseases or follow similar phenotypic outcomes. Mice are thus used in several different disease models that can be later translated to human conditions including asthma, COPD, toxicants, cystic fibrosis, as well as viral and bacterial infections. The standard histological embedding orientation of the lung provides an easy view of the main bronchus, alveoli, bronchioles, trachea, and related lymph nodes for distinct research inquiries. However, the standard histologi

BENEFITS of Tissue-Tek Paraform Sectionable Cassette System Authors: Jason (Jay) Innerhofer, PA(ASCP), M.H.S., Albuquerque, NM The Paraform System eliminates the need to perform manual steps at embedding and great in terms of training, histology and turn around time.

Leveraging preprocessor scanning to improve safety, quality, and lean workflow in the anatomic pathology lab. Authors: Lilly Guevara, Kristie Wolfe-Steele, Brian Johnson While procedures exist in AP laboratories to protect patient samples and prevent loss through specimen tracking, misplaced cassettes between grossing and tissue processing are still problematic. Significant time may be spent locating a misplaced cassette in this part of the workflow. Pathologists Bio-Medical (PBM) Pathgroup implemented a new system to address this need for preprocessing sample tracking in their AP workflow. PBM worked to create their own solution for tracking AP samples. This included the use of barcodes, scanners, and high-resolution cameras to track samples throughout the lab. It was a time consuming and costly effort pursued over the course of years. However, visibility in the pre-processing workflow was still problematic. In some instances, cassettes scanned at grossing were not used at all, or a cassette was inadvertentl

The Button Mushroom (Agaricus bisporus): An Alternative Control Source for the Grocott Methenamine Silver Technique Authors: From University of Texas M.D. Anderson Cancer Center Center, Houston, TX- Fatimah Ansar; Carla Arredondo; Tran Huynh; Victoria Jones; Jennifer Le; Duong Nguyen; Andres Ronquillo-Erazo; Minnu Varghese; Kaleena Ramirez; Toysha Mayer; Mark A. Bailey   Identifying the presence of fungi through special histotechniques is a critical diagnostic histopathology test for patients who may have acquired a fungal infection localized in the lungs. The purpose of this study was to determine if the mushroom species Agaricus bisporus, generally known as the Button Mushroom and, commonly used for cooking, may be used a special stain control slide. Is the Button Mushroom a reliable fungal control to use when performing a Grocott Methenamine Silver technique test?  The investigators hypothesize the Agaricus bisporus mushroom is a good substitute control for result verification  of the GMS staining techniqu

 Gene Protein Assay(GPA): Unveiling Tumor Heterogeneity in HER2/neu Positive Breast Cancer Authors:  Dr. Tanuja Shet, Dr. Aditi Rathi    Introduction: HER2/neu gene amplification on Fluorescence in-situ hybridization (FISH) is defined as average HER2neu gene signals > 4 and HER2:CEP17 ratio of > 2, in breast cancer. However, with tumor heterogeneity interpretation of these cut-offs is challenging and some cases test as borderline. We attempted a study analyzing Gene protein assay (GPA) which combines HER2/neu immunohistochemistry (IHC), and D-DISH (dual color dual in-situ hybridization) assay to help resolve this issue.    Methods:  A total of 31 cases reported as HER2/neu amplified with intermixed tumor heterogeneity on FISH were taken for this study. The original HER2/neu count was between 4 to 6 and ratio near 2 in most cases. GPA was done using 4B5 HER2/neu IHC and Roche/Ventana D-DISH kit on Ventana Ultra machine by doing IHC first followed by D-DISH. Result:  On GPA, 3/31 cases were reported as no