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Annals of Oncology : Official Journal... Jan 2019
Topics: Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunotherapy; Microdissection; Squamous Cell Carcinoma of Head and Neck; Tumor Microenvironment; Virtual Reality
PubMed: 30476183
DOI: 10.1093/annonc/mdy516 -
Current Protocols in Neuroscience Jan 2019The heterogeneous organization of the mammalian neocortex poses a challenge for elucidating the molecular mechanisms underlying its physiological processes. Although...
The heterogeneous organization of the mammalian neocortex poses a challenge for elucidating the molecular mechanisms underlying its physiological processes. Although high-throughput molecular methods are increasingly deployed in neuroscience, their anatomical specificity is often lacking. In this unit, we introduce a targeted microdissection technique that enables extraction of high-quality RNA and proteins at high anatomical resolution from acutely prepared brain slices. We exemplify its utility by isolating single cortical columns and laminae from the mouse primary somatosensory (barrel) cortex. Tissues can be isolated from living slices in minutes, and the extracted RNA and protein are of sufficient quantity and quality to be used for RNA sequencing and mass spectrometry. This technique will help to increase the anatomical specificity of molecular studies of the neocortex, and the brain in general, as it is applicable to any brain structure that can be identified using optical landmarks in living slices. © 2018 by John Wiley & Sons, Inc.
Topics: Animals; Coloring Agents; Mice; Microdissection; Neocortex; Nerve Net; Skull; Somatosensory Cortex
PubMed: 30285322
DOI: 10.1002/cpns.55 -
PloS One 2016Precision medicine promises to enhance patient treatment through the use of emerging molecular technologies, including genomics, transcriptomics, and proteomics....
Precision medicine promises to enhance patient treatment through the use of emerging molecular technologies, including genomics, transcriptomics, and proteomics. However, current tools in surgical pathology lack the capability to efficiently isolate specific cell populations in complex tissues/tumors, which can confound molecular results. Expression microdissection (xMD) is an immuno-based cell/subcellular isolation tool that procures targets of interest from a cytological or histological specimen. In this study, we demonstrate the accuracy and precision of xMD by rapidly isolating immunostained targets, including cytokeratin AE1/AE3, p53, and estrogen receptor (ER) positive cells and nuclei from tissue sections. Other targets procured included green fluorescent protein (GFP) expressing fibroblasts, in situ hybridization positive Epstein-Barr virus nuclei, and silver stained fungi. In order to assess the effect on molecular data, xMD was utilized to isolate specific targets from a mixed population of cells where the targets constituted only 5% of the sample. Target enrichment from this admixed cell population prior to next-generation sequencing (NGS) produced a minimum 13-fold increase in mutation allele frequency detection. These data suggest a role for xMD in a wide range of molecular pathology studies, as well as in the clinical workflow for samples where tumor cell enrichment is needed, or for those with a relative paucity of target cells.
Topics: Animals; Cell Nucleus; Epithelium; High-Throughput Nucleotide Sequencing; Humans; Mice; Microdissection; NIH 3T3 Cells; Staining and Labeling
PubMed: 26999048
DOI: 10.1371/journal.pone.0151775 -
Pathology Aug 2022Solid-type adenoid cystic carcinomas (ACCs) are highly aggressive and heterogeneous tumours. Because of their rarity, therapeutic strategies guided by genetic profiles...
Solid-type adenoid cystic carcinomas (ACCs) are highly aggressive and heterogeneous tumours. Because of their rarity, therapeutic strategies guided by genetic profiles based on next generation sequencing (NGS) have not been published for these tumours. Forty-nine solid-type ACCs including 43 tumours with a predominantly solid pattern, and six tumours comprising a roughly equal mixture of cribriform/tubular and solid histological forms were included in our study. The solid components from the 49 solid ACCs were enriched for mutations of genes in the NOTCH pathway (NOTCH1 61%, SPEN 24%) and chromatin remodelling pathway and the absence of myoepithelial cell differentiation. Cases with NOTCH1 mutations exhibited strong NICD expression, which was associated with poor overall and distant metastasis free survival. BRCA2 mutation and BCOR/BCORL1 mutations were observed in 20% and 18.4% of solid ACCs, respectively. In six of the solid ACCs, intratumour heterogeneity was delineated between the cribriform/tubular and solid components. NOTCH1 and FGFR2 mutations as well as NOTCH2 amplification were restricted to the solid component, indicating clonal selection within the same tumour. In two recurrent/metastatic solid ACCs, the subclones evolved in progression for local relapse and distant metastasis, although they manifested close genomic resemblance to primary tumours. Guided by the genetic profiles, the preclinical efficiency of the gamma-secretase inhibitor BMS-906024 was evaluated in patient derived xenograft models (PDXs) with activating NOTCH1 mutations and demonstrated robust antitumour effects. Our report revealed intratumour heterogeneity among solid-types within an ACC as well as the inter-tumour evolution of dominant clones among two primary and recurrent/metastatic tumours. In contrast to cribriform/tubular ACCs, solid-type ACCs should be approached with a distinct therapeutic strategy, particularly targeting NOTCH1. Microdissecting the highest grade component guided by histology is a highly recommended tumour sampling strategy and facilitates the detection of key molecular targets.
Topics: Carcinoma, Adenoid Cystic; Genetic Heterogeneity; Humans; Immunohistochemistry; Microdissection; Neoplasm Recurrence, Local
PubMed: 35337667
DOI: 10.1016/j.pathol.2021.12.292 -
Journal of Oral and Maxillofacial... Mar 2020Trigeminal neuralgia is a chronic and debilitating syndrome characterized by short paroxysms of lancinating facial pain. Patients may be medically managed; however, in...
Trigeminal neuralgia is a chronic and debilitating syndrome characterized by short paroxysms of lancinating facial pain. Patients may be medically managed; however, in cases of medically refractory trigeminal neuralgia, surgical management is often required. Our objective was to present and describe a technique for endoscopic microdissection of the infraorbital nerve, a peripheral method of management for refractory V2 trigeminal neuralgia in patients without evidence of neurovascular compression. The technique is designed to spare sensation in unaffected portions of the V2 distribution. We present 2 patients with medically refractory V2 trigeminal neuralgia localized to the lateral midface who underwent infraorbital microdissection. After first confirming that there was no neurovascular compression on imaging in these patients, we administered infraorbital bupivacaine injections to localize the symptomatic nerve. The nerve was then accessed via a 1.5-cm buccogingival incision, and the connective tissue sheath was incised. The nerve fascicles were bluntly separated, and the symptomatic branches were cauterized with fine-tipped monopolar cautery. Both patients reported complete resolution of their pain postoperatively and were pain free at last follow-up. They reported some hypoesthesia in the lateral face; however, they retained some sensation in the medial upper lip, midface, and nose. Infraorbital microdissection is a safe and effective technique for symptomatic management of V2 trigeminal neuralgia while sparing sensation in asymptomatic portions of the dermatome.
Topics: Endoscopy; Humans; Maxillary Nerve; Microdissection; Pain; Trigeminal Nerve; Trigeminal Neuralgia
PubMed: 31751521
DOI: 10.1016/j.joms.2019.10.011 -
MSphere Jan 2019The highly oncogenic alphaherpesvirus Marek's disease virus (MDV) causes immense economic losses in the poultry industry. MDV induces a variety of symptoms in infected...
The highly oncogenic alphaherpesvirus Marek's disease virus (MDV) causes immense economic losses in the poultry industry. MDV induces a variety of symptoms in infected chickens, including neurological disorders and immunosuppression. Most notably, MDV induces transformation of lymphocytes, leading to T cell lymphomas in visceral organs with a mortality of up to 100%. While several factors involved in MDV tumorigenesis have been identified, the transformation process and tumor composition remain poorly understood. Here we developed an imaging mass spectrometry (IMS) approach that allows sensitive visualization of MDV-induced lymphoma with a specific mass profile and precise differentiation from the surrounding tissue. To identify potential tumor markers in tumors derived from a very virulent wild-type virus and a telomerase RNA-deficient mutant, we performed laser capture microdissection (LCM) and thereby obtained tumor samples with no or minimal contamination from surrounding nontumor tissue. The proteomes of the LCM samples were subsequently analyzed by quantitative mass spectrometry based on stable isotope labeling. Several proteins, like interferon gamma-inducible protein 30 and a 70-kDa heat shock protein, were identified that are differentially expressed in tumor tissue compared to surrounding tissue and naive T cells. Taken together, our results demonstrate for the first time that MDV-induced tumors can be visualized using IMS, and we identified potential MDV tumor markers by analyzing the proteomes of virus-induced tumors. Marek's disease virus (MDV) is an oncogenic alphaherpesvirus that infects chickens and causes the most frequent clinically diagnosed cancer in the animal kingdom. Not only is MDV an important pathogen that threatens the poultry industry but it is also used as a natural virus-host model for herpesvirus-induced tumor formation. In order to visualize MDV-induced lymphoma and to identify potential biomarkers in an unbiased approach, we performed imaging mass spectrometry (IMS) and noncontact laser capture microdissection. This study provides a first description of the visualization of MDV-induced tumors by IMS that could be applied also for diagnostic purposes. In addition, we identified and validated potential biomarkers for MDV-induced tumors that could provide the basis for future research on pathogenesis and tumorigenesis of this malignancy.
Topics: Animals; Biomarkers, Tumor; Chickens; Image Processing, Computer-Assisted; Isotope Labeling; Laser Capture Microdissection; Lymphoma; Marek Disease; Proteome; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 30651403
DOI: 10.1128/mSphere.00569-18 -
Andrology May 2017
Topics: Azoospermia; Humans; Male; Microdissection; Sperm Retrieval; Treatment Outcome
PubMed: 28409905
DOI: 10.1111/andr.12356 -
Folia Morphologica 2020In this study, we explored the specific microanatomical properties of the trigeminal ganglion (TG) blood supply and its close neurovascular relationships with the...
BACKGROUND
In this study, we explored the specific microanatomical properties of the trigeminal ganglion (TG) blood supply and its close neurovascular relationships with the surrounding vessels. Possible clinical implications have been discussed.
MATERIALS AND METHODS
The internal carotid and maxillary arteries of 25 adult and 4 foetal heads were injected with a 10% mixture of India ink and gelatin, and their TGs subsequently underwent microdissection, observation and morphometry under a stereoscopic microscope.
RESULTS
The number of trigeminal arteries varied between 3 and 5 (mean 3.34), originating from 2 or 3 of the following sources: the inferolateral trunk (ILT) (100%), the meningohypophyseal trunk (MHT) (100%), and from the middle meningeal artery (MMA) (92%). In total, the mean diameter of the trigeminal branches was 0.222 mm. The trigeminal branch of the ILT supplied medial and middle parts of the TG, the branch of the MHT supplied the medial part of the TG, and the branch of the MMA supplied the lateral part of the TG. Additional arteries for the TG emerged from the dural vascular plexus and the vascular network of the plexal segment of the trigeminal nerve. Uniform and specific intraganglionicdense capillary network was observed for each sensory trigeminal neuron.
CONCLUSIONS
The reported features of the TG vasculature could be implied in a safer setting for surgical approach to the skull base, in relation to the surrounding structures. The morphometric data on TG vasculature provide anatomical basis for better understanding the complex TG blood supply from the internal and external carotid arteries.
Topics: Aged; Cadaver; Female; Fetus; Humans; Male; Microdissection; Middle Aged; Trigeminal Ganglion
PubMed: 31282551
DOI: 10.5603/FM.a2019.0062 -
Laboratory Investigation; a Journal of... Jul 2017Analysis of specific DNA alterations in precision medicine of tumors is crucially important for molecular targeted treatments. Lung cancer is a prototypic example and...
Analysis of specific DNA alterations in precision medicine of tumors is crucially important for molecular targeted treatments. Lung cancer is a prototypic example and one of the leading causes of cancer-related deaths worldwide. One major technical problem of detecting DNA alterations in tissue samples is cellular heterogeneity, that is, mixture of tumor and normal cells. Microdissection is an important tool to enrich tumor cells from heterogeneous tissue samples. However, conventional laser capture microdissection has several disadvantages like user-dependent selection of regions of interest (ROI), high costs for dissection systems and long processing times. ROI selection in expression-based microdissection (xMD) directly relies on cancer cell-specific immunostaining. Whole-slide irradiation leads to localized energy absorption at the sites of most intensive staining and melting of a membrane covering the slide, so that tumor cells can be isolated by removing the complete membrane. In this study, we optimized xMD of lung cancer tissue by enhancing staining intensity of tumor cell-specific immunostaining and processing of the stained samples. This optimized procedure did not alter DNA quality and resulted in enrichment of mutated EGFR DNA from lung adenocarcinoma specimens after xMD. We here also introduce a quality control protocol based on digital whole-slide scanning and image analysis before and after xMD to quantify selectivity and efficiency of the procedure. In summary, this study provides a workflow for xMD, adapted and tested for lung cancer tissue that can be used for lung tumor cell dissection before diagnostic or investigatory analyses.
Topics: Adenocarcinoma; Adenocarcinoma of Lung; DNA; Formaldehyde; Humans; Immunohistochemistry; Lung; Lung Neoplasms; Microdissection; Molecular Diagnostic Techniques; Mutation; Staining and Labeling; Tissue Fixation
PubMed: 28436954
DOI: 10.1038/labinvest.2017.31 -
Frontiers in Endocrinology 2022Nonobstructive azoospermia (NOA) is a common and severe form of male infertility. Microdissection testicular sperm extraction (microTESE) combined with intracytoplasmic...
INTRODUCTION
Nonobstructive azoospermia (NOA) is a common and severe form of male infertility. Microdissection testicular sperm extraction (microTESE) combined with intracytoplasmic sperm injection (ICSI) is an optimal treatment for men with NOA. However, the outcomes and affecting factors of ICSI for NOA patients with different etiologies receiving microTESE treatment are still unclear.
METHODS
A total of 335 NOA patients undergoing microTESE from January 2017 to December 2021 were included in this retrospective analysis. The patients were divided into five groups (idiopathic, Klinefelter syndrome (KS), Y chromosome microdeletions (YCMDs), cryptorchidism and mumps orchitis) according to the etiologies. The clinical characteristics and outcomes of microTESE and ICSI were collected and comparisons were performed between clinical characteristics of patients who had successful sperm retrieval (SSR) and sperm retrieval failure (SRF). In addition, relationships between clinical characteristics and rates of SSR were explored by Kendall correlation analysis.
RESULTS
The overall SSR rate was 40.90%. SSR rate of the idiopathic group (31.22%) was the lowest and was much lower than that of other groups (KS: 48.65%, 28/58; YCMDs: 60.87%; cryptorchidism: 80.95%; mumps orchitis: 75.00%). The overall fertilization rate was 72.26%. No group differences were found among five groups (idiopathic: 73.91%; KS: 71.43%; YCMDs: 64.29%; cryptorchidism: 70.59%; mumps orchitis: 77.78%). The overall clinical pregnancy rate was 66.67%. No group differences were found among five groups (idiopathic: 68.63%; KS: 65.00%; YCMDs: 44.44%; cryptorchidism: 66.67%; mumps orchitis: 85.71%). The overall live birth rate was 66.67%. No group differences were found among five groups (idiopathic: 71.43%; KS: 53.85%; YCMDs: 50.00%; cryptorchidism: 75.00%; mumps orchitis: 66.67%). For SSR patients, the average age was significantly lower in the idiopathic group, while the average testicular volume was significantly greater in the cryptorchidism and mumps orchitis groups. However, no significant differences were found in the level of follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone (T) between patients who had SSR and SRF. In addition, negative relationships were found between age and rates of SSR in idiopathic NOA patients while positive relationships were found between testis volume and rates of SSR in patients with cryptorchidism and mumps orchitis.
CONCLUSION
Patients with idiopathic NOA had lowest SSR. In addition, the age in idiopathic NOA patients was a predictor for SSR while testicular volume in NOA patients with cryptorchidism and mumps orchitis was a predictor for SSR. However, the relationships between clinical characteristics and clinical outcomes in NOA patients were preliminary, and further validation needed to be carried out in a larger sample to increase statistical capacity before a definitive conclusion could be drawn.
Topics: Pregnancy; Female; Humans; Male; Azoospermia; Microdissection; Sperm Injections, Intracytoplasmic; Cryptorchidism; Orchitis; Retrospective Studies; Mumps; Semen; Spermatozoa
PubMed: 36325443
DOI: 10.3389/fendo.2022.1006208