HistoTalks: NSH Podcasts

NSH Poster Podcast: P08/P11 (2024)

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 the recently introduced UK NEQAS CPT Diagnostic Cytopathology Cell block scheme will be presented and how this information can be circulated to laboratories experiencing difficulties with their preparation methods as an advisory service. This will include.
• Understanding the clinical importance and diagnostic purpose of correct procedures in Diagnostic Cytopathology Cell Block preparation.
• Identifying and determining factors affecting best practice and quality in Diagnostic Cytopathology Cell Block practice and how to resolve them.
• Identifying and understanding the causes of artefacts experienced in Diagnostic Cytopathology Cell Block preparation methods and how to eliminate and prevent them.

P11- Participating In A Digital Diagnostic Cytopathology Interpretive Proficiency Testing (iEQA) Scheme -Helen Naylor, Anna Patterson, Chantell Hodgson, Ashley Makela

The iLabXCell Digital Diagnostic Cytopathology Interpretive Proficiency Testing Scheme or iEQA is facilitated by UK NEQAS Cellular Pathology Technique (CPT), and is designed to promote quality, excellence, and education for all involved in screening and reporting of Diagnostic Cytopathology. It is open to medical and non-medical staff, trainees, advanced practitioners, and staff associated with Cytopathology. iEQA provides superior, outstanding, and representative case examples for individuals to examine in a remote setting and submit an opinion via the user-friendly digital platform. 
Offering 2 circulations annually it provides
• Easy access for registration 
• Ability to choose specimen types 
• Advanced slide viewing
• Clinical details to assist diagnoses
• Categorisation of images using benign or malignant 

Flexibility
Remote learning allows participants to continue learning alongside their peers, in an environment accommodating their needs. It allows the flexibility of international participants to access the platform at any time, convenient to them. Remote learning via the iEQA platform, offers the flexibility for users to pick up learning where they left it – anywhere, anytime, from any location.
Self-paced learning
Traditional learning has long failed to acknowledge the individualised nature of learning, opting instead for a generalised approach that may not be optimised for everyone. One of the many benefits of this iEQA is providing participants more independence to:
• Spend more time on cases/case types they find difficult
• Revisit the images as often as they need 

Environmental benefits
Digitising slides has led to a decrease in physical resources previously necessary for iEQA to function. Everything participants need is accessible on-line, eliminating the historical environmental impact and the time taken for circulations to be distributed and completed.  

Future developments
An on-line educational image library, containing images of Diagnostic Cytopathology cases of urines, serous fluids, respiratory and head and neck cases used in previous circulations for educational and training purposes. 

 

 

 

 

NSH Poster Podcast: P06/ P15 (2024)

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;

Can the EM unit produce good quality sections and images that maximizes the chances of observing relevant pathologies?
Data from UK NEQAS CPT show that only 50% of EM units participating in the diagnostic TEM scheme
achieve excellent scores of 9 or 10 out of 102 not achieving excellent quality could potentially compromise a diagnosis.
The knowledge the staff have of ultrastructural pathology
• Do the staff assessing your samples know what to look for? Having proof of EM staff’s knowledge in ultrastructural
pathology is essential when relying on them to provide relevant images and a considered report on features seen
or not seen.

Questions to Ask Your EM Provider
To ensure that the EM service you are sending your samples to is ‘fit for purpose’, you should not only consider the speed and cost of the service but also;
• Ensure that your potential provider is accredited to ISO 15189 standards.
• Ask for evidence of participation in an EQA scheme specifically for technical TEM.
• Ask for TEM EQA results over the past 12 months and ensure they are consistently achieving excellent marks.
• Do the EM staff participate in regular knowledge and competence competency review specifically for TEM
and ultrastructural assessment?
• How much experience of ultrastructural pathology do the members of staff examining your samples have
and do they have any qualifications in this area?
• Get endorsements from other users of the EM service to evidence the quality of work offered.

Summary

Access to EM services remains vital across the globe, but in the UK is increasingly being outsourced to units remote from the originating trust. In this case, the pathologist is reliant on the images and ultrastructural report being accurate to inform diagnosis.
To ensure accuracy, the EM unit, whether they be UK based or part of our international community, all should participate in a quality EQA scheme and all staff should be experienced and have access to training to ensure
they are educated to a high level in ultrastructural pathology.
Without this, the referring trust cannot be guaranteed the service they are paying for is fit for purpose.

P15-Understanding the Quality of your Electron Microscopy Provider in this Era of Outsourcing of Services: How does a Technical EQA Scheme Add Value? -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 Benefit of Participation in an EQA Scheme  Summary The Transmission Electron Microscopy (TEM) Scheme was developed in 2018 as a means of assessing the quality of the final output of diagnostic TEM Units against a defined assessment standard. The aim of this EQA scheme is to allow EM units to have their work regularly assessed and measured against other TEM units in an anonymous and supportive manner. This ensures that results are reliable and comparable no matter where they are performed. Education in technical aspects of the TEM process is also offered as a knowledge and competence exercise as part of each EQA assessment. EQY Scheme Organization TEM scheme Participants are asked to submit 4 digital images from each of 2 contrasted TEM cases. There are 6 EQA assessment runs per year. Specific tissue types for each case are requested for each assessment run. Renal cases are requested for each run as case 1 and muscle or nerve are requested in rotation for case 2. However alternative tissue types can be submitted for either case if participants do not examine those tissue types. Details of technical fixation, processing and imaging for each case are required to be submitted as part of Data Entry. This allows generation of ‘best method’ reports for high achieving EQA scores to be issued to Participants. Each image is anonymously assessed against the defined assessment  criteria by a pair of expert peer assessors. Each assessor will award a score out of 5 giving a total score for each image out of 10. Participants receive the following for each run; • EM Individualized report detailing the scores awarded for each image, along with information about where marks were lost and why • EM best method report detailing methods that achieved high scores • EM image report giving examples of images that achieved a range of scores from the maximum score to low scoring images, along with why those scores were allocated  Summary Participation in a technical TEM EQA scheme provides factual information that can also be used to advertise and promote an EM unit as providing a proven quality service.  Whilst being participating in an educational exercise within each EQA assessment, allows individual and laboratory knowledge and competence to be assess and demonstrated. Evidencing your EM unit provides a high quality, gold standard service is vital when diagnostic electron microscopy is often being outsourced to external EM units. Cost of service and turn round times for results are often asked, but quality of work for diagnostic results should also be considered. Consistency of technical quality is important to ensure pathologists receive high quality diagnostic information. This mitigates against misinterpretation from poor quality images, assuring a safe, accurate diagnosis. This is good news for the participating EM units, good news for the diagnostic process and ultimately, good news for patients.

NSH Poster Podcast: P37 (2024)

Research Requires Flexibility: Protease-Free Permeabilization Expands FISH Tissue Applications.-Andrelie Branicky, Shared Laboratory Resources, Lerner Research Institute, Cleveland Clinic, Cleveland, OH

Fluorescence in situ hybridization (FISH) visualizes the presence of a specific DNA or RNA sequence in a tissue sample or cell. This method, particularly the mRNA version, detects gene expression when protein might not be present or IHC is impossible. FISH combined with immunohistochemistry enables spatial transcriptomics, which provides significantly more information about the tissue microenvironment.

Formalin-fixed paraffin-embedded (FFPE) tissues are the standard for tissue preservation in the clinical world. Most commercial mRNA probe and amplification systems are built around the model of FFPE tissue that can withstand harsh protease permeabilization. In the research world, tissues are fixed in different fixatives for varying times; all at the discretion of the investigator instead of an organization like the CLIA. 

Given the wide range of tissue preparations, the HCR automated FISH-ISH protease-free program provides the flexibility to combine  FISH and fluorescent immunohistochemistry on tissue fixed in a variety of ways such as: 10% NBF, Histochoice (a glyoxal-based fixative), and methanol/acetic acid, with only minor changes to the basic protocol. Additionally, the lack of harsh protease pre-treatment maintains tissue integrity and morphology for staining and imaging. 

NSH Poster Podcast: P35 (2024)

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 differences can allow for easy distinction. This is why a cocktail run with a dual staining technique would be a superior diagnostic tool in comparison to testing the two markers independently.

NSH Poster Podcast: P36 (2024)

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 catered to individuals without access to an accredited educational program. Information has been organized in ways that focus on areas that may be particularly difficult, information heavy or exam relevant. They utilize multiple learning modalities, specifically written, spoken, and visual information retrieval methods, with opportunities for auditory and group learning. They are also intended to be cost effective. Materials include laminated study templates, laminated scratch paper, flashcards, a slideshow for visual tissue assessment, and a general study topic list. Questions are both simple and complex, and representative of exam questions. It should be noted, however, that this is not a comprehensive study guide and is intended to be used in addition to an online study program or exam preparation course and textbook resources. These tools are incomplete guides which can be added to and personalized.

 

NSH Poster Podcast: P46 (2024)

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 reagents.

NSH Poster Podcase: P54 (2024)

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 platforms. Although the process is laborious and requires continued refinement the slides should be sufficient for highplex proteomics and transcriptomics methods and can be used in addition to FFPE tissues for research into immunology, oncology, and virology.

NSH Poster Podcast: P51 (2024)

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, minimizing the need to recut and restain the sample to ensure proper tissue morphological characteristics. Adhesion slides may also be used for H&E stains and special stains for added adhesion, but could retain excess reagent, or background staining, on the slide. In this study the differences in contact angle and in tissue adherence during microtomy were analyzed,  investigation was done on whether different adhesion slides exhibit similar levels of background staining during histological staining procedures, and evaluation and comparison of the tissue adhesion properties of adhesion slide brands across different tissue types and applications was performed. 
After wide-ranging testing of adhesion slide characteristics, this study exhibited that not all adhesion slides are created equal. While water bath behavior showed to not be a relevant factor, there was considerable variation in background staining and tissue adhesion between slides. The results of this study suggested that it is important to determine what the needs are for your laboratory based on the types of staining done and tissue types used. It is also important to test adhesion slides to find the right slide or slides for your laboratory applications.

NSH Poster Podcast: P32 (2024)

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 mutation in all cancer cases, aligning with results from other genetic testing methods, and as expected, non-cancerous cases showed no mutation-specific staining. The study highlighted important considerations in interpreting IHC results, such as the presence of non-specific brown staining due to colloid containing brown pigments in negative cases. It also emphasized the importance of proper tissue representation, as demonstrated by one case that initially lacked staining. While the research provides strong evidence for the accuracy of the Ventana anti-BRAFV600E antibody, it also identified limitations. The study lacked PTC cases negative for the BRAFV600E mutation, which would have provided a more comprehensive validation. Conclusion: The researchers suggest further investigation with a broader range of samples, including more anaplastic thyroid carcinoma cases and follicular variant of PTC, to explore potential correlations between tumor heterogeneity and staining intensity. This validation study contributes to improving thyroid cancer diagnosis and treatment by authenticating the reliability IHC methods in detecting a key genetic mutation. It also highlights the importance of continued research to optimize diagnostic methods in thyroid cancer.

NSH Poster Podcast: P17 (2024)

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, we maintained a frozen NHP brain as a single piece while coronally sectioning through the entire cerebral hemispheres followed by cerebellum/brain stem. The brain remained frozen on a sliding microtome by utilizing a freezing stage and molds filled with powdered dry ice. As the brain was sectioned at 40um the molds were removed to uncover additional tissue for cutting until the tissue had been entirely sectioned. Serial pictures and an organized series of well plates were used to preserve anatomical order. By sectioning the brain as a whole and maintaining control over anatomical context we achieved results that may enhance the accuracy and thoroughness of histopathological examinations. Furthermore, serial sectioning of the Paraformaldehyde-fixed frozen brain preserved the antigenicity for various immunohistochemical assays. We believe this technique offered certain advantages over the routine paraffin-embedded sectioning method, especially for the biodistribution studies practiced during the developmental stage of gene therapies.

NSH Poster Podcast: P14 (2024)

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 well as their docility, capacity for repeated blood draws, and ability to tolerate numerous implants during a single operation. In this study, sheep were implanted with devices containing allogeneic primary fetal sheep or xenogeneic Rat-2 fibroblasts to determine cell survival and immune response. Methods: Primary fetal sheep fibroblasts were isolated and cultured in RPMI. Rat-2 fibroblasts (ATCC) were cultured in DMEM. Cells were loaded into devices at different densities and transplanted subcutaneously. Sheep underwent allogeneic or xenogeneic transplantation with empty control and perforated devices transplanted as well. Devices were explanted at days 3, 7, 21 and 38 to evaluate host responses, vascularization, and cell survival through histological evaluation. Results: Sheep fibroblasts survived at different densities, confirming alloprotection with good vascularization and absence of a major foreign body response or extensive inflammation around the implants. Devices overloaded at the highest densities exhibited some host immune responses. Perforated devices exhibited a greater immune response that infiltrated the device. A robust immune response was seen around xenogeneic transplants with no encapsulated cells surviving. Conclusion: Sheep make a valuable translational animal model in cell therapy research. The results of this study concluded alloprotection in immunoisolating devices, allorejection in purposely perforated devices, and host immune response to xenogeneic transplantation, representing a feasible model for translational development of cell therapies. Future work will include continued allogeneic studies while avoiding xenogeneic rejection by incorporating immunomodulatory technologies.

NSH Poster Podcast: P12 (2024)

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 immunohistochemistry (IHC) protocols for reliable localization the PgCad1 receptor in cultured Tni insect cell lines. Such protocols may be useful for in situ localization of wild type PgCad1 and mutant variants in midgut cells of the pink bollworm.

 

 

Presenting at the NSH Convention: Our Histology Community is Amazing!

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. 

Presenting Advice for the NSH Convention: Just Do It!

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!

NSH Poster Podcast: P033 (2023)

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 examination, it was discovered the CytoLyt (Thinprep) specimen was not being processed because of lack of specimen. When it was processed the slides were being dried to long leading to artifact on the slide. By increasing the sample size in radiology and using a slide dryer to regulate the drying, the lab was able to produce a more consistent result. 

Following these changes, the lab projected to meet or exceed the goal of less than 10% non-diagnostic slide results by the end of 2023.

NSH Poster Podcast: P031 (2023)

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 histological orientation may not demonstrate the specific areas of interest to further investigate these conditions. By trimming the lung lobes in specific ways or altering the orientation during embedding, we can provide a specific focus to a study.  One alternative embedding strategy is the “max airway”, which focuses on showing the main bronchus anatomy on all lung lobes. The “morphometry” embedding strategy allows for multiple cross-sections of the left lung lobe and a cross-section of the main airway.  Another common method is the “lung sampling” strategy which provides a systematically chosen cross-section of the lungs. A “left lobe focus” strategy is used to review the bronchioles and alveolar areas. Finally, “whole lung” embedding can be useful for examining general lung morphology, adjacent mediastinal structures, and other organs. Using these different protocols, we can bring a desired focus to a study and have a more scalable experiment without using human tissue early in the process.

NSH Poster Podcast: P024 (2023)

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.

NSH Poster Podcast: P020 (2023)

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 inadvertently returned to the specimen container. In order to truly advance patient safety, improve quality, and optimize lean workflow, this needed to be addressed. In March 2023, PBM implemented a system that automated data capture for cassettes at preprocessing. This resulted in a reduction in time associated with reconciling misplaced/missing cassettes from grossing to embedding: from as long as a few days to as short as a couple hours. Another benefit of the new system was tracking cassettes by processor retort. This is especially important for laboratories that have multiple dual retort processors supporting various tissue types. In the past, lab staff spent significant time solving processing quality issues and identifying which cassettes were loaded on which processor(s). The new system software enables full reporting of this information. This system has provided a quick method to investigate processing errors with various specimen types. These changes created a more efficient laboratory, reduced stress for staff, and enhanced turnaround times.

NSH Poster Podcast: P011 (2023)

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 technique and readily available compared to tissue controls. The button mushrooms were purchased from a local grocer in Houston, TX, subsequently grossed into eight 1.0x1.0x0.4cm representative sections and submitted for routine tissue processing, embedding, sectioning and stained using the H&E and Grocott Methenamine methods. Inclusive, for the test a commercially available human lung tissue control (+) for fungi was compared to the Button Mushroom results and to ensure the GMS technique produced the appropriate result. The processed button mushrooms GMS stain did not yield a definitive result to conclude the outcome of the staining method. The stained GMS slides demonstrated an indistinguishable contrast between the morphology of the mushroom and the presence of fungi; however, the H&E demonstrated the morphology well, however, further studies are necessary to confirm the presence of fungi.   The use of alternative controls are dependent on the specific clinical; research goals or experimental conditions to provide appropriate and relevant results. The limitations of the research was, not together with, the  GMS to perform a PAS stain on the button mushrooms. Subsequently, we will perform the PAS technique on the button mushrooms and test a different alternative source to establish a viable control for the substitution of human tissue (+) for fungi.  

NSH Poster Podcast: P007 (2023)

 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 non-amplified, and the rest were amplified. Advantage of GPA was that it helped count D-DISH as per membrane staining and identified more tumor heterogeneity in contrast to FISH in six cases. The results of the remaining cases were the same.

NSH Poster Podcast: P006 (2023)

Novel Use of Pipette Based Capsules for Glycol Methacrylate, Methyl/Butyl Methacrylate and Epoxy Resin Processing with Mouse Tissue That Reduce Processing Solution Volume and Time.  

Authors: Philip Seifert, HTL(ASCP), Xinyao Hu, Bianai Fan and Darlene A. Dartt, PhD, Schepens Eye Research Institute of Mass Eye and Ear, Boston, MA

 

The mPrep/S capsules (Microscopy Innovations) are customized pipette tips with screen inserts designed for various electron microscopy applications. Capsule processing methods were developed for mouse ocular and mineralized bone for histology and transmission electron microscopy (TEM) applications and compared to manual and carousel processed methods. The novel methods tested enable processing larger sized tissues (~3 mm diameter) including whole, anterior or posterior segments of mouse eyes and optic nerves within a single capsule that use 75% less reagents and time. Capsule processed tissue embedded in Glycol Methacrylate, methyl methacrylate/butyl methacrylate and EMBED812 epoxy resin were sectioned for histological staining including H&E, toluidine blue, osmium-paraphenylenediamine, Von Kossa and TEM imaged. Stained slide scanned micrographs and TEM images from capsule processed samples were measured and compared to conventional processed using a carousel processor or manual processing.  There were no detectable differences in tissue preservation, staining, optic nerve axon density or processing related artifacts in any of the tissue processed into the resins.  Use of capsules with pipette propelled mixing delivers accelerated diffusion of reagents and infiltration of embedding media into specimens that decreases both processing volume and time compared to manual or carousel processing.  A variety of embedding media resins may be used with capsules for histology and electron microscopy tissue processing. 

AwardsCast: 2023 Immunohistochemistry Education Scholarship

Join us for our AwardsCast series as we have short conversations with the 2023 NSH Award & Scholarship Recipients. In this episode, we talk to Colleen Forster of the University of Minnesota, the recipient of the Immunohistochemistry Education Scholarship, where we discuss how every day in a core research lab is different and keeps her learning new things as well as her plans for learning more about the hot new area of spatial omnics.

AwardsCast: 2023 Jules Elias Award for Excellence in IHC & Molecular Techniques

Join us for our AwardsCast series as we have short conversations with the 2023 NSH Award & Scholarship Recipients. In this episode, we talk to Cindy Lowe, HTL(ASCP), QIHC(ASCP), of Vanderbilt University, the recipient of the Jules Elias Award for Excellence in IHC & Molecular Techniques, where we discuss everything from her work in IHC, including an impactful visit from patients to her lab, to what she does for fun outside the lab.

AwardsCast: 2023 Helping Hand Scholarship for Histotechnology Outreach and Education

Join us for our AwardsCast series as we have short conversations with the 2023 NSH Award & Scholarship Recipients. In this episode, we talk to Beth Cox, CT (ASCP), HT(ASCP), HTL (ASCP),
QIHC (ASCP), the recipient of the Helping Hands Scholarship for Histotechnology Outreach & Education where she discusses how she will use the funds to help identify and treat gynecologic cancer in women in the Marshall Islands. 

AwardsCast: 2023 Peggy Wenk Histology Program Scholarship

Join us for our AwardsCast series as we have short conversations with the 2023 NSH Award & Scholarship Recipients. In this episode, we talk to Program Director, Taiquanda Winbush, from Albany State University about how they will use the funds to improve their program with the funds from the scholarship.

AwardsCast: 2023 Excellence in Management Award

Join us for our AwardsCast series as we have short conversations with the 2023 NSH Award & Scholarship Recipients. In this episode, we talk to Thomas Huynh, HT(ASCP) who gives his advice on what it takes to be a good manager and the importance of having a good mentor.

AwardsCast - 2023 Histotechnologist of the Year

Join us for our AwardsCast series as we have short conversations with the 2023 NSH Award & Scholarship Recipients. In this episode, we talk to Histotechnologist of the Year, Luis Chiriboga, PhD, HT(ASCP), QIHC(ASCP) and we discuss the importance of advocacy and volunteering.

 

 

NSH Poster Podcast: P-09 (2022)

Title: Using Metallic Labels and Metal Detection in the Histology Laboratory to Prevent Missing Material

Authors: Elizabeth Druffel, HT ASCP, Mayo Clinic, Rochester, MN; Dr. Joaquin Garcia, MD, Mayo Clinic, Rochester MN

NSH Poster Podcast: P-13 (2022)

Title: Anatomical Pathology Patient Interest Association (APPIA) TOPS Tissue Handling Guidelines illustrate an ongoing education initiative to improve the understanding of pre-analytical factors and promote standardization in tissue workflows.

Authors: Robert Lott, HTL(ASCP), Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC; Joshua Greenlee, MBA IASSC, HTL(ASCP)cm, Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC; Melissa Alexander, MD PhD, Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC; Jodi Barrientos, Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC; David Ferber, MD, Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC; Beth Sheppard, MBA, Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC; Shalini Singh, MD, Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC; Teri Saylor, Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC; Shanya Scott, Anatomical Pathology Patient Interest Association, Raleigh, NC; Erico von Bueren, MD PhD MOR, Anatomical Pathology Patient Interest Association (APPIA), Raleigh, NC

NSH Poster Podcast: P-26 (2022)

Title: Detection of Human Papillomavirus (HPV) mRNA in FFPE Samples Using AMPIVIEW RNA Probes developed with Novel loop Technology

 

Authors: Deborah M. Holzapfel, PhD, Francesca Mazzoni, PhD, Jack Coleman, PhD

NSH Poster Podcast: P-01 (2022)

Title: Comparative Study of Two Reprocessing Methods for Formalin Fixed Paraffin Embedded Tissue

 

Authors: Matthew Lunetta BS HT(ASCP)cm, Cameron Felty DO, Megan Grivois BS HT(ASCP)cm, Chris Hansen HT(ASCP), Chris Jackson MD, and Peter Seery BS PA(ASCP)cm , Liz Rizzo PA(ASCP)cm LSSBB 

NSH Poster Podcast: P-03 (2022)

Title:  Histological assessment of C1q expression and deposition at the neuromuscular junction (NMJ) in Tibialis Anterior (TA) Tissue of SOD1G93A Mouse Model

 

Authors: Joseph Vereen, MS, Annexon Biosciences, Brisbane, CA
Alessia Tassoni, PhD, Annexon Biosciences, Brisbane, CA
Larry Mattheakis, PhD, Annexon Biosciences, Brisbane, CA
Ellen Cahir-McFarland ,PhD, Annexon Biosciences, Brisbane, CA
Ted Yednock, PhD, Annexon Biosciences, Brisbane, CA
Yaisa Andrews-Zwilling, PhD, Annexon Biosciences, Brisbane, CA

 

What is Mentorship?

In this episode, NSH members Ashley Carter and Colleen Wisnieski talk about their experiences with mentorship.  Each explain how they have personally been involved with mentorship and by doing so help to better define how mentor-mentee relationships work.

Celebrating Histotechnologists in the Lab - Pamela Vizcarra

This podcast celebrates histotechnologists as part of Medical Laboratory Professionals Week. In this series, we celebrate histotechs who work in a variety of different settings including research, clinical, and veterinary labs and a medical examiner's office. 

In this episode, Pamela Vizcarra discusses how she started out as a lab assistant and was able to receive on-the-job training to become a histology supervisor in a research lab doing clinical trials. 

Clelebrating Histotechnologists in the Lab - Peter Socki

This podcast celebrates histotechnologists as part of Medical Laboratory Professionals Week. In this series, we celebrate histotechs who work in a variety of different settings including research, clinical, and veterinary labs and a medical examiner's office. 

In this episode, Peter Socki explains how his love of crime shows as a child lead to him becoming a histotechnologist working in a medical examiner's office.

Celebrating Histotechnologists in the Lab - Sean Moynihan

This podcast celebrates histotechnologists as part of Medical Laboratory Professionals Week. In this series, we celebrate histotechs who work in a variety of different settings including research, clinical, and veterinary labs and a medical examiner's office. 

In this episode, Sean Moynihan discusses how he applies his degree in biology with a focus on nuclear medicine to what he does on a day-to-day basis working in a dermatopathology outreach lab.

Celebrating Histotechnologists in the Lab - Katelin Murphy

This podcast celebrates histotechnologists as part of Medical Laboratory Professionals Week. In this series, we celebrate histotechs who work in a variety of different settings including research, clinical, and veterinary labs and a medical examiner's office. 

In this episode, Katelin Murphy discusses how she became interested in histology through a recommendation by her high school guidance counselor and now works in a veterinary pathology lab. 

Celebrating Histotechnologists in the Lab - Jennifer Betustak

This podcast celebrates histotechnologists as part of Medical Laboratory Professionals Week. In this series, we celebrate histotechs who work in a variety of different settings including research, clinical, and veterinary labs and a medical examiner's office. 

In this episode, Jennifer Betustak explains how she went from being a phlebotomists to becoming a Histology Supervisor.

 

Celebrating Histotechnologists in the Lab - Janet Tunnicliffe

This podcast celebrates histotechnologists as part of Medical Laboratory Professionals Week. In this series, we celebrate histotechs who work in a variety of different settings including research, clinical, and veterinary labs and a coroner’s office. 

In this episode, we speak with Janet Tunnicliffe about her experience in British Columbia working as a histotech and she gives thoughtful advice to those considering Histotechnology as a career.  

Women in Histology 2021 - Haydee Lara Histology Outreach

In this episode NSH members David Krull and Haydee Lara, Cellular Biomarker Investigator with GSK discuss Ms. Lara's incredible outreach work with STEM programming, her own journey into science and histology, and what is next for her career.  

Women in Histology 2021 - Faith Rice Falloposcope

In this episode Michelle Bell interviews Faith Rice, Senior Research Specialist from the University of Arizona to discuss Falloposcope, a project that will potentially save lives but providing early detection for ovarian cancer.  They also discuss working in research.  To learn more about this project, click here.

Published:

Kelli C. Kiekens, Gabriella Romano, Dominique Galvez, Ricky Cordova, John Heusinkveld, Kenneth Hatch, William Drake, Zaynah Kmeid, Jennifer K. Barton"Re-engineering a Falloposcope Imaging System for Clinical Use", Translational biophotonics, Volume2, Issue4, November 2020, e202000011.

Women in Histology 2021 - Dr. Elisa Balducci DYRK1A

In this episode Dr. Elisa Balducci and Andrea Transou discuss Dr. Balducci's research journey with DYRK1A, her transition from research to industry, and her STEM outreach work.  

AwardsCast: Automation, Standardization, and Validation

In this episode of NSH’s podcast series AwardsCast, Nadia Gale, Region IX Director and member of the IHC Committee talks to Jeremy Johnston, who happens to be Region VIII Director and the 2020 recipient of the Excellence in Standardization of IHC scholarship. They talk about all things IHC; everything from how automation has impacted standardization, to the definition of companion diagnostics, to which antibodies are giving them grief during the validation process. Applications for the 2021 Excellence in Standardization of IHC Scholarship, sponsored by Agilent Dako, are open through June 4^th. Apply today!

 

AwardsCast: Visualizing Inflammasome Structure

In this episode of AwardsCast, the 2020 recipient of the Newcomer Helping Hand Award, Dr. Regina Vontell, sits down with Konnie Zeitner, NSH Speaker of the House and fellow past award recipient, to discuss Dr. Vontell’s work with visualizing inflammasome structure in the brain. Applications for the 2021 Newcomer Helping Hand Award, sponsored by Newcomer Supply, are open until June 4th. Apply today! 

 

AwardsCast: Histotech of the Year

In this episode of AwardsCast, LaVinia Ray, the 2020 recipient of the Histotech of the Year Award, talks about how she has grown in her career at Southern Research, where she is currently working on COVID-19 studies. She describes the benefits of NSH volunteerism, working 20 years for one institution, and how she feels about being one of the great histotechs honored with this most prestigious NSH award. Applications are now open for the 2021 Histotech of the Year Award, sponsored by Epredia. Nominate today! 

 

AwardsCast: The Philippines

In this episode of AwardsCast, Evelyn Diaz, the 2020 recipient of the Lee Luna Foreign Travel Scholarship, and Jomar Klee Custodio, the President of the Philippine Society for Histotechnology, discuss Evelyn’s plans to use her scholarship to return to her country of origin, the Philippines, to assist the PSH in their mission of raising the bar for histotechnology. Applications for the 2021 Lee Luna Foreign Travel Scholarship, sponsored by Sakura Fintek, USA, are open through June 4th. Apply today! 

 

AwardsCast: Reproducibility in Scientific Research

In this episode of AwardsCast, Gayle Callis, Editor in Chief of the Journal of Histotechnology, meets with Luis Chiriboga, Region I Director, and the 2020 recipient of the Editor’s Award, a unique NSH award given by the Journal Editor for outstanding accomplishments in the realm of publications. They talk about the importance of reproducibility in scientific research and the steps JOH has taken to improve its standing in the scientific research community.

 

HistoHelp Podcast: AI in the Lab - Part 2

Welcome to HistoHelp, the podcast series filled with tips and tricks for the lab.  The great advice in this series is provided by NSH members who are here to help you with your common...and not so common problems! In Part 2 of this episode NSH member David Krull and Adam Smith, Indica Labs continue their discussion about AI in the lab and dive deeper into multiplexing and challenges with implementation. ,

HistoHelp Podcast: AI in the Lab - Part 1

Welcome to HistoHelp, the podcast series filled with tips and tricks for the lab.  The great advice in this series is provided by NSH members who are here to help you with your common...and not so common problems! In Part 1 of this episode NSH member David Krull invites Adam Smith, Indica Labs to talk about AI in the lab, discuss validation, understanding data, and ways you can implement AI/digital in the lab. 

HistoHelp Podcast: Bielschowsky Stain

Welcome to HistoHelp, the podcast series filled with tips and tricks for the lab.  The great advice in this series is provided by NSH members who are here to help you with your common...and not so common problems! In this episode, NSH member Eric Peterson really does come across a not so common problem involving polar bear tissue.