Fixation Of Specimens Research Articles

Organocatalyst treatment improves variant calling and mutant detection in archival clinical samples

Formalin fixation of biological specimens damages nucleic acids and limits their use in genomic analyses. Previously, we showed that RNA isolation with an organocatalyst (2-amino-5-methylphenyl phosphonic acid, used to speed up reversal of formalin-induced adducts) and extended heated incubation (ORGΔ) improved RNA-sequencing data from formalin-fixed paraffin-embedded (FFPE) tissue samples. The primary goal of this study was to evaluate whether ORGΔ treatment improves DNA-sequencing data from clinical FFPE samples. We isolated RNA and DNA ± ORGΔ from paired FFPE and frozen human renal and ovarian carcinoma specimens collected as part of the National Cancer Institute Biospecimen Pre-analytical Variables program. Tumor types were microscopically confirmed from adjacent tissue sections. Following extraction, DNA was fragmented and sequenced and differences were compared between frozen and FFPE sample pairs. Treatment with ORGΔ improved concurrent SNP calls in FFPE DNA compared to non-ORGΔ FFPE samples and enhanced confidence in SNP calls for all FFPE DNA samples, beyond that of matched frozen samples. In general, the concordant SNPs identified in paired frozen and FFPE DNA samples agreed for both genotype and homozygosity vs. heterozygosity of calls regardless of ORGΔ treatment. The increased confidence in ORGΔ FFPE DNA variant calls relative to the matched frozen DNA suggests a novel application of this method. With further optimization, this method may improve quality of DNA-sequencing data in FFPE as well as frozen tissue samples.

Influence of Manual and Ultrasonic Scaling on Surface Roughness of Four Different Base Materials Used to Elevate Proximal Dentin–Cementum Gingival Margins: AnIn VitroStudy

To evaluate and compare the effects of both manual and ultrasonic scaling on surface roughness of four different base materials, used for elevating dentin/cementum gingival margins of proximal cavities. Eighty human upper molars with compound Class II mesial cavities, with gingival margins 1 mm below the cemento-enamel junction (CEJ), were divided into four different groups according to the type of the base material used; resin-modified glass ionomer (RMGI), glass hybrid (HV-GIC), flowable bulk-fill resin composite (Bulk Flow) and bioactive ionic resin (Activa). This was followed by completing the restorations with the same resin composite. All materials were used according to the manufacturers' instructions. All groups were further subdivided into two subgroups according to the scaling technique: manual (hand) or ultrasonic. All restorative and scaling procedures were performed after fixation of specimens with acrylic beside neighboring teeth to simulate natural contact. The mean surface roughness (Ra, μm) of all specimens was measured quantitatively and qualitatively by a three-dimensional (3D) surface analyzer system at two stages; (1) after thermal cycling for 5000 cycles without scaling and (2) after scaling. Data were statistically analyzed using analysis of variance (ANOVA), Tukey post hoc tests, and paired sample t-tests (at α=0.05). For baseline readings, the Bulk Flow group had the lowest Ra values, while HV-GIC group had the highest. RMGI and Activa groups had no statistical significant difference between their Ra values (p>0.05). For post scaling readings, hand scaling had significantly lower Ra values than ultrasonic scaling in all the material groups (p<0.05), except in the Bulk Flow group, where both scaling methods were not significantly different from each other (p>0.05). Bulk Flow had the smoothest surfaces when cured against a matrix band compared with the other tested base materials. When hand and ultrasonic scaling methods were compared, the latter technique had more detrimental effect on the surface texture of the four tested base materials.

Improvement of the specimen fixation method before t-butyl alcohol freeze-drying for SEM observation of the hymenium of Pleurotus ostreatus

Improvement of the specimen fixation method before t-butyl alcohol freeze-drying for SEM observation of the hymenium of <i>Pleurotus ostreatus</i>

Perfusion-distention fixation of heart specimens: a key step in immortalizing heart specimens for wax infiltration and generating 3D imaging data sets for reconstruction and printed 3D models

Perfusion-distention fixation by gravity is a fundamental cardiac predissection technique that may be used to preserve the 3D anatomy of a cardiac specimen. The equipment and steps are illustrated, and examples of cardiac specimens preserved in various methods including wet specimens, wax-infiltrated hearts, and 3D-printed models, created after performing this technique, are shown.

Introduction to Molecular Diagnostics of Insects

Insects play an important role in biocenoses due to their abundance and wide (cosmopolitan) distribution. Many insects are crop pests. An effective pest control could be realized in case of proper species identification, which is usually managed by morphological analysis. Molecular methods allow to deep study of many issues of insect biology. In particular, traditional approach can not ordinary identify a species at all stages of their life cycle, whereas molecular methods can it. This review covers a wide range of issues related to the molecular genetic analysis of insects. In the first section we consider the methods of fixation and storage of insect specimens, as well as their impact on DNA quality. Further, we provide general information on population study design. Various schemes of DNA extraction, examples of both express techniques and more thorough protocols for DNA extraction and their purification are provided. In addition, methods of DNA isolation that allow to preserve a specimen integrity for further morphological studies are considered. The methods of DNA quality control are described in detail, that is important for PCR analysis. The last section provides various methods of PCR analysis, that we exemplify by studies aimed to elucidate both fundamental issues and practical problems.

Surgico-pathological analysis of testicular lesions

The primary objective of our study was to identify the different types of histopathological lesions of testes and epididymis occurring in patients admitted in a tertiary care hospital, the objective was also to analyze the spectrum of testicular lesions, our aim was to evaluate the prevalence of testicular malignancies and characterize the tumor based on their histo-morphological features. This was a prospective study of 2 years (October 2018 to October 2020) aggregating a total of 52 cases. The surgical specimens were referred to the pathology department for histopathological analysis. Adetailed preoperative history of every case with regards to age, presenting symptoms, and signs along with personal history were obtained.Also, relevant investigations like serum tumor markers and Ultrasonography findings were analyzed. Postoperatively, the tissue was sent to the histopathology department was studied by proper fixation, gross examination, adequate sampling, and paraffin sections of the same.10% formalin was used to fix the specimen. Some resources suggest the use of Bouin solution for the fixation of testis biopsy specimens. However, the picric acid in the Bouin solution presents challenges for safe handling and disposal due to its explosive potential.Hence, given the available resources in our laboratory setting, formalin was the preferred choice of fixative. We ensured that the specimens came in adequate amount of formalin from the OT with correct labeling.We excluded the autolyzed specimens. The data was noted and quantitative analysis was done.The study was further compared with other previously reported studies. 52 cases were studied and majority of the cases were reported in the age group of 31-40yrs. 65.4% cases were non neoplastic, with testicular torsion being the commonest lesion. Amongthe 23.1% neoplastic cases that were reported, mixed germ cell tumor was the commonest (75%). Non neoplastic lesions are more common than neoplastic lesions. Torsion testis was the commonest indication for orchidectomy in younger age group. Despite newer techniques in imaging and tumor marker assays the diagnosis of testicular lesions is primarily dependent upon histopathological examination.

Postmortem brain donations vs premortem surgical resections for glioblastoma research: viewing the matter as a whole.

There have been limited improvements in diagnosis, treatment, and outcomes of primary brain cancers, including glioblastoma, over the past 10 years. This is largely attributable to persistent deficits in understanding brain tumor biology and pathogenesis due to a lack of high-quality biological research specimens. Traditional, premortem, surgical biopsy samples do not allow full characterization of the spatial and temporal heterogeneity of glioblastoma, nor capture end-stage disease to allow full evaluation of the evolutionary and mutational processes that lead to treatment resistance and recurrence. Furthermore, the necessity of ensuring sufficient viable tissue is available for histopathological diagnosis, while minimizing surgically induced functional deficit, leaves minimal tissue for research purposes and results in formalin fixation of most surgical specimens. Postmortem brain donation programs are rapidly gaining support due to their unique ability to address the limitations associated with surgical tissue sampling. Collecting, processing, and preserving tissue samples intended solely for research provides both a spatial and temporal view of tumor heterogeneity as well as the opportunity to fully characterize end-stage disease from histological and molecular standpoints. This review explores the limitations of traditional sample collection and the opportunities afforded by postmortem brain donations for future neurobiological cancer research.

MRI before biopsy correlates with depth of invasion corrected for shrinkage rate of the histopathological specimen in tongue carcinoma

The purpose of this study was to evaluate which radiological depth of invasion (r-DOI) measurement is the most concordant to clinical DOI (c-DOI) derived from correction for the shrinkage rate of the histopathological specimens. We retrospectively reviewed 128 patients with tongue carcinoma who had undergone glossectomy between 2006 and 2019. At first, the width shrinkage rate during formalin fixation and preparation process of histopathological specimens was evaluated. From the shrinking rates, a formula to calculate c-DOI from pathological DOI (p-DOI) was developed. The correlation between c-DOI and r-DOI was evaluated. The specimen shrinkage rate during the histopathological specimen preparation process was 10.3%. Based on that, we yielded the correct formula for c-DOI based on p-DOI and preparation shrinkage rate: c-DOI = p-DOI × 100/89.7. The regression equations for the association of c-DOI with r-DOI measured by ultrasound (n = 128), MRI before biopsy (n = 18), and MRI after biopsy (n = 110) were y = 1.12 * x + 0.21, y = 0.89 * x − 0.26, and y = 0.52 * x + 2.63, respectively, while the coefficients of determination were 0.664, 0.891, and 0.422, respectively. In conclusion, r-DOI using MRI before biopsy most strongly correlated with c-DOI.

Olfactory bulb SARS-CoV-2 infection is not paralleled by the presence of virus in other central nervous system areas.

Olfactory bulb SARS-CoV-2 infection is not paralleled by the presence of virus in other central nervous system areas.

Fixation Using Carnoy's Solution Enables Detection of More Lymph Nodes After Gastrectomy for Gastric Cancer.

To compare the number of lymph nodes (LNs) detected when using Carnoy's solution (CS) versus 10% neutral buffered formalin (NBF) to fix specimens after radical gastrectomy for gastric cancer. LNs were routinely detected using NBF until 2020, since then, for the fixation procedure, residual fat was fixed in CS for 24 hours and dissected again for the detection of further LNs. Of 143 specimens, 117 were included in the NBF group and 26 in the CS group. The mean numbers of LNs examined were 27.85±14.89 and 36.30±12.41 in the NBF and CS groups, respectively (p=0.008). The mean number of additional LNs detected using CS was 8.07±2.91, of which 0.38±1.02 were metastatic. Additional LNs were found in all patients of the CS group, and all were ≤3 mm. Of the 26 patients in the CS group, metastatic LNs were detected in four, disease in two of whom was up-staged. CS is an appropriate alternative to NBF for the fixation of gastric cancer specimens, and more LNs were detected in the resected specimens fixed when using CS compared with NBF.

3D printed clamps for fixation of spinal segments in biomechanical testing

3D printed clamps provide multiple advantages compared to potting for the fixation of spinal specimens and in a recent study, superior fixation stability was reported. The aim of this study was to evaluate the fixation efficacy of 3D printed vertebra clamps during routine application and to present and evaluate a novel clamp for sacrum fixation. Further, public access to the template files is provided. 98 human single-level cadaveric specimens were biomechanically tested in flexion–extension (FE), lateral bending (LB), axial rotation (AR), anteroposterior shear (AS), lateral shear (LS) and axial compression-decompression (AC). Loading amplitudes were +/-7.5 Nm for FE, LB and AR, +/- 150 N for AS and LS and + 400/-100 N for AC. The novel sacrum clamp was used in 8 specimens. The median relative motion between clamps and specimens was 0.6° in FE, 0.7° in LB, 0.3° in AR, 0.5 mm in AS, 0.5 mm in LS and 0.1 mm in AC. With sacrum clamps, the median relative motion was 0.3° in FE, 0.1° in LB, 0.08° in AR, 0.8 mm in AS, 0.7 mm in LS and 0.2 mm in AC. The vertebra clamps used during routine testing provided better stability compared to the values in the literature in all six loading directions (p < 0.05). The sacrum clamp showed superior anchoring stability in three loading directions compared to the caudal vertebra clamps (p < 0.05), while inferior stability was measured in AS (p < 0.001). We conclude that 3D printed vertebra clamps and 3D printed sacrum clamps represent reliable methods for specimen fixation during routine biomechanical testing.

Consensus recommendations for the standardized histopathological evaluation and reporting after radical oesophago-gastrectomy (HERO consensus).

Variation in the approach, radicality, and quality of gastroesophageal surgery impacts patient outcomes. Pathological outcomes such as lymph node yield are routinely used as surrogate markers of surgical quality, but are subject to significant variations in histopathological evaluation and reporting. A multi-society consensus group was convened to develop evidence-based recommendations for the standardized assessment of gastroesophageal cancer specimens. A consensus group comprised of surgeons, pathologists, and oncologists was convened on behalf of the Association of Upper Gastrointestinal Surgery of Great Britain & Ireland. Literature was reviewed for 17 key questions. Draft recommendations were voted upon via an anonymous Delphi process. Consensus was considered achieved where >70% of participants were in agreement. Consensus was achieved on 18 statements for all 17 questions. Twelve strong recommendations regarding preparation and assessment of lymph nodes, margins, and reporting methods were made. Importantly, there was 100% agreement that the all specimens should be reported using the Royal College of Pathologists Guidelines as the minimum acceptable dataset. In addition, two weak recommendations regarding method and duration of specimen fixation were made. Four topics lacked sufficient evidence and no recommendation was made. These consensus recommendations provide explicit guidance for gastroesophageal cancer specimen preparation and assessment, to provide maximum benefit for patient care and standardize reporting to allow benchmarking and improvement of surgical quality.

Introduction of a New Pathology Workup Protocol for Glottic Cancer Treated With Transoral Laser Microsurgery (TLM): Prospective Analysis of Oncological Outcomes and Matched Case-Control Study.

Background/PurposeThe value of margin status after TLM for glottic cancer is debatable, due to difficulties in specimen orientation and margin analysis. To reduce these difficulties, we recently introduced a standardized protocol of oriented fixation of TLM specimens. This proved feasible and resulted in high margin evaluability rates and a decreased rate of false positive deep margins, when compared to a historical TLM cohort. For the patients whose specimens were processed according to this protocol, we prospectively analyzed oncological outcomes, identified prognostic factors and assessed the influence of the protocol introduction on outcomes compared with a historical TLM cohort.MethodsNinety-six patients with glottic malignancies treated with TLM were included. Resection specimens were processed according to the new protocol. Descriptive statistics and survival analyses were used to determine oncological outcomes. To assess the effect of the protocol introduction on outcomes, a matched-case-control analysis was performed, using a historical TLM-cohort as controls. The Cox proportional hazards model was used to analyze prognostic effects of patient and treatment characteristics, including the pathology protocol introduction, on overall survival (OS), disease-specific survival (DSS), disease-free survival (DFS) and local recurrence-free survival (LRFS).ResultsTwo-year outcomes were favorable: 88.5% OS, 97.0% DSS, and 87.6% LRFS. At multivariable analysis, the presence of multiple positive superficial margins was a negative prognosticator for OS (HR 4.102) and increasing cT classification proved a negative prognosticator for DFS (HR 2.828) and LRFS (HR 2.676). Matched case-control analysis did not reveal a significant difference in oncological outcomes between cohorts. Deep margin status had a strong differential effect for DFS (p-value for interaction = 0.0205) and for LRFS (p-value for interaction = 0.0176) between cohorts, indicating a prognostic effect of deep margin status on both outcomes in the current cohort, but not in the historical cohort.Discussion/ConclusionThe introduction of a new standardized technique of oriented fixation of TLM specimens did not affect oncological outcomes when compared to a historical TLM cohort, but assigned a significant prognostic effect to deep margin status for DFS and LRFS, facilitating the decision making process with regards to planning of second-look procedures, administration of adjuvant radiotherapy or determination of follow-up intensity.

Guidelines for removal, preservation, and CT imaging of the syrinx, the avian vocal organ

While the state of the art has been described for recording bird songs and calls, there are no described best practices for collecting and preserving the avian vocal organ, the syrinx. In addition to skins and skeletal preparations, field collection of tissues for DNA sequencing has become a common practice. Protocols for such tissue collection often involve accessing internal organs. Here we argue that collecting syringes, which also involves accessing the chest cavity, adds only minimal time to field or skeletal preparation and can be done without damage to the skeleton. Collecting syringes will enable studies of structure–function relationships in the production of calls and song that have not been previously possible. Three-dimensional X-ray computed tomography (CT) imaging techniques allow new insight into these relationships, but material of both male and female birds as well as juveniles is lacking for most avian taxa. Here, we present a guide to the removal and preservation of the avian syrinx and associated vocal anatomy in hopes of increasing their representation in biological collections. In this guide, we suggest that taking in situ measurements is ideal since dissection, fixation, and tissue staining can affect measurements of certain airway features such as tracheal diameter and bronchial length. Our guidelines demonstrate that removing a single wing, shoulder, and half of the pectoral girdle from the dorsal side of the thorax along with disarticulating the sternal and vertebral rib segments on one side of the body allows exposure of the complete airway without damaging the skeleton. We also provide recommendations for fixation, storage, and staining of specimens in preparation for diffusible iodine-based contrast-enhanced computed tomography (diceCT) scans that allow imaging of both soft tissue structures such as vocal folds and muscles as well as the cartilaginous rings that support these structures.

Developing of the optimal shape and reinforcement structure of the specimen for adequate determination of the tensile strength in unidirectional composites

Unidirectional composites exhibit the highest strength when stretched along the fibers. However, the proper determination of the strength faces great methodological difficulties. The main problems of tensile testing of polymer composites consisted in developing of the specimen shape and the method of specimen fixation which ensure the minimum impact of the stress concentration near the grips on the strength measurements. A conventional shape of the specimen with fillets is unsuitable for unidirectional polymers due to the splitting occurred in the fillet zones upon loading. Therefore, the specimens are usually standardized in the form of rectangular strips fixed using pads or special grips which provide constant transverse forces. However, with such a specimen shape, a significant stress concentration inevitably occurs at the edge of grips and the lower the ratio of the interlayer shear modulus to the longitudinal Young’s modulus, the greater the stress concentration impact. For the purpose of the most correct determination of the strength we propose to use specimens with smoothly varying dimensions at the same cross-sectional area which ensures keeping the total number of unbroken fibers in each section. The specimen thickness decreases when moving from the working part of the specimen to the gripping part, whereas the width (while maintaining the section area) grows to prevent the specimen collapsing resulting from transverse forces in standard self-tightening grips. Analytical and FEM modeling is performed to select a rational contour shape. Technological equipment has been developed and a procedure of manufacturing testing specimens has been worked out. The tensile test of specially manufactured curvilinear reinforced specimens showed higher strength values compared to standard rectangular strips or specimens with semicircular fillets.

Use of scopolamine butylbromide to facilitate specimen fixation after endoscopic submucosal dissection.

Endoscopic submucosal dissection (ESD) is a technique for the en bloc resection of early neoplastic lesions in the gastrointestinal tract. A step that must be carried out after excision is the pinning down of the specimen on a support plate such as a cork board to avoid artifacts because of tissue contraction. After ESD lesion borders usually curl up from contraction, hence this procedure must be carefully performed to prevent damaging the specimen. Recently, Nishizawa T, et al. reported that dropping adrenaline onto the lesion results in edge relaxation, which facilitates fixation. However, the use of other drugs-such as Buscapine-that induce gastrointestinal smooth muscle relaxation, potentially with similar effects, has not been described. Below, two ESD specimens from two different institutions are shown. These specimens correspond to gastric lesions. Buscapine is applied on their surface and the edges become relaxed, thus facilitating stretching and thus fixation. In conclusion, the use of Buscapine may help in the processing of excised lesions by facilitating pinning out and reducing damage risk for specimens.

Spectrum of gastrointestinal polyps: A tertiary care hospital experience of five years

Introduction: Polyps of gastrointestinal tract are not uncommon specimens received in surgical pathology laboratory. With advent of advanced endoscopy it is now possible to precisely sample polyps and determine its nature; weather benign, malignant or with malignant potential. Present study was performed with aim to study incidences of various histological types of polyps of gastrointestinal tract, to study its clinical presentation and to study polyps in relation to their size and site, in a tertiary care hospital. Materials and Methods: A retrospective and prospective analysis of 189 cases of gastrointestinal polyps was carried out over a period of eight and half years. Relevant clinical details of patient, size, number, size and presence of stalk etc. were noted. Specimens were studied for gross and microscopic examination after fixation of specimen with 10% formalin and microscopically examining H & E slides. They were classified according to site and various histological types. Results: Overall, we analysed 189 cases of gastrointestinal polyps. Commonest site was large intestine and commonest type was Juvenile polys, followed by neoplastic polyps, hyperplastic polyps, inflammatory pseudo polyps, hamartomatous polyps, inflammatory fibroid polyps and 2 cases of Familial polyposis coli were note. Conclusion: Present study reveals that gastrointestinal tract had wide spectrum of non-neoplastic as well as neoplastic polyps. They have different clinical significances and therapeutic implications. Histopathology play pivotal role in arriving at final diagnosis which enables to study detail incidences and helps to differentiate non-neoplastic diseases with clinical presentation of malignancy. Incidence of various polyps in based on institutional data and may not reflect incidence of state or nation. Keywords: Gastrointestinal polyps, Adenomatous polyps, Juvenile polyps, Neoplastic polyps.

Transoral Laser Microsurgery (TLM) for Glottic Cancer: Prospective Assessment of a New Pathology Workup Protocol.

Background/Purpose: The value of margin status after TLM for glottic cancer is debatable, due to difficulties in specimen orientation and margin analysis. Purpose of this study was the prospective evaluation of feasibility of a new standardized technique of oriented fixation of the TLM specimen and identification of the added value on tissue processing and margin status reporting.Methods: Patients with suspicious glottic lesions undergoing TLM were included. After resection, the specimen margins were inked in the OR using different colors. Subsequently, the specimens were fixed on a pig liver carrier and sent for further processing, accompanied with photographs of the larynx pre-TLM and of the mounted specimen. Feasibility was assessed by registration of duration of specimen preparation in the OR and the lab and by procedure-specific questionnaires. Objective evaluation included assessment of margin status and proportion of evaluable margins. Chi square tests were used to make comparisons of proportions.Results: One hundred and four consecutive patients were included between May 2016 and September 2019. TLM was performed in a primary and salvage setting in 89.4 and 10.6% of patients, respectively. Mean duration of intraoperative specimen preparation was 5.1 min (SD 2.6 min). No difficulties in orientation nor fixation during intraoperative preparation were reported in 87.5 and 88.2%, respectively. Specimen orientation was judged by the pathologist as very adequate in 89.4%, with the accompanying photographs considered helpful for orientation and processing in 84.6%. Substantial difficulties in further lab processing and pathologic examination were identified in 17.7%. Deep margin evaluability was very high (98.0%) and significantly higher than the evaluability of superficial mucosal margins. Compared to our previous series published by our group (n = 142), deep margin evaluability significantly rose from 62.7 to 98.0% (p < 0.001) and true positive rate of the deep margins increased from 0 to 44.4% (p = 0.002).Discussion/Conclusion: The new and standardized technique of oriented fixation of TLM specimens on a pig liver carrier proves feasible both in the OR and lab setting and results in high margin evaluability rates, especially for the deep margin, as well as a decreased rate of false positive deep margins when compared to a historical TLM cohort.

The Immunohistochemical Expression of Programmed Death Ligand 1 (PD-L1) Is Affected by Sample Overfixation.

Humanized antibodies targeting programmed death receptor 1 (PD-1) or its ligand (PD-L1) have been approved for the treatment of different cancers. Some of these antibodies show a correlation between the tissue expression of PD-L1 and response. Evaluation of PD-L1 expression presents multiple challenges, but some preanalytical issues such as tissue fixation have been scarcely evaluated. With the hypothesis that immunohistochemical staining of PD-L1 may be impacted by the time of specimen fixation, we evaluated differences in its expression in tonsil samples exposed to predefined fixation times. Random nontumoral tonsillectomy specimens were blindly evaluated in tissue microarray slides after staining with SP142 and SP263 antibodies. With fixation times ranging from 12 to 72 hours, between 2.8% and 6.1% of the samples were considered to be suboptimally stained, with no differences between the 2 antibodies within these fixation times. A significantly higher proportion of samples exposed to a fixation time of 96 hours presented suboptimal immunostaining (15.6%, P<0.0001). In addition, suboptimally stained spots were 20.8% using SP142 and 10.4% using SP263 after 96 hours of fixation (P=0.046). In conclusion, the quality of staining for PD-L1 in tonsil samples decreased with overfixation of the specimen at times >72 hours. Samples exposed to formaldehyde for longer periods presented suboptimal results for both clones, but the SP142 antibody presented a significantly lower tolerance to formalin overexposure than SP263. These results indicate the relevance of a controlled preanalytical processing of samples and particularly the length of fixation of tumor specimens.

Best Practices Guideline for the Pathologic Diagnosis of Breast Implant-Associated Anaplastic Large-Cell Lymphoma.

To provide guidelines for the accurate pathologic diagnosis of breast implant-associated anaplastic large cell lymphoma (BIA-ALCL), the preoperative evaluation of the patient with suspected BIA-ALCL, and the pathologic evaluation of the capsulectomy specimen. To better inform patients and healthcare providers about BIA-ALCL, we convened to review diagnostic procedures used in the evaluation of patients with suspected BIA-ALCL. We focused on the processing of the seroma fluid/effusion surrounding the implant, the handling of capsulectomy specimens following removal of implant(s), and the preoperative evaluation of the patient with suspected BIA-ALCL. Recommendations were based on the published literature and our experience to optimize procedures to obtain an accurate diagnosis and assess for tumor invasion and the extent of the disease. Early diagnosis of BIA-ALCL is important as the disease can progress and deaths have been reported. Because the most common presentation of BIA-ALCL is swelling of the breast with fluid collection, an accurate diagnosis requires cytologic evaluation of the effusion fluid surrounding the affected implant. The first priority is cytocentrifugation and filtration of fresh, unfixed effusion fluid to produce air-dried smears that are stained with Wright-Giemsa or other Romanowsky-type stains. Preparation of a cell block is desirable to allow for hematoxylin and eosin staining and immunohistochemical analysis of formalin-fixed, paraffin-embedded histologic sections. Cell block sections can be used for polymerase chain reaction-based investigation of T-cell receptor gene rearrangement to detect clonality. Fixation and mapping of the capsulectomy specimen to select multiple representative sections are advised to assess for microscopic tumor involvement and capsular invasion. It is appropriate to assess lymph node involvement by excisional biopsy material rather than fine needle aspiration, due to propensity for focal involvement.