Recent developments in multiplexing techniques for immunohistochemistry

Dixon, Angela R.; Bathany, Cédric; Tsuei, Michael; White, Joshua B.; Barald, Kate F.; Takayama, Shuichi

doi:10.1586/14737159.2015.1069182

Cited by 103 publications

(74 citation statements)

References 87 publications

(157 reference statements)

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“…The combination of mass spectroscopy with IHC has allowed multiplexed, direct quantitative imaging of tissue samples for basic and clinical research 73. Multiplexed IHC methods permit identification of at least three, and up to 30 discrete antigens 74. Some studies suggest that IHC may be sufficiently sensitive and specific to predict response to targeted therapy 75…”

Section: Protein-based Detection Methodsmentioning

confidence: 99%

Detection approaches for multidrug resistance genes of leukemia

Chen

2017

DDDT

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Leukemia is a clonal malignant hematopoietic stem cell disease. It is the sixth most lethal cancer and accounts for 4% of all cancers. The main form of treatment for leukemia is chemotherapy. While some cancer types with a higher incidence than leukemia, such as lung and gastric cancer, have shown a sharp decline in mortality rates in recent years, leukemia has not followed this trend. Drug resistance is often regarded as the main clinical obstacle to effective chemotherapy in patients diagnosed with leukemia. Many resistance mechanisms have now been identified, and multidrug resistance (MDR) is considered the most important and prevalent mechanism involved in the failure of chemotherapy in leukemia. In order to reverse MDR and improve leukemia prognosis, effective detection methods are needed to identify drug resistance genes at initial diagnosis. This article provides a comprehensive overview of published approaches for the detection of MDR in leukemia. Identification of relevant MDR genes and methods for early detection of these genes will be needed in order to treat leukemia more effectively.

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Section: Protein-based Detection Methodsmentioning

confidence: 99%

Detection approaches for multidrug resistance genes of leukemia

Chen

2017

DDDT

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“…This field is currently taking a huge leap forward due to advances in digital pathology and due to the rapid development of simultaneous multispectral analysis of many different markers in one small biopsy, which will ultimately lead to its implementation in day-to-day pathology practice (Feng et al , 2015). Therefore, objective, high-throughput biomarker quantification and co-localisation using multiplexed IHC will aid in the future selection of patients that might benefit from specific treatment (Huang et al , 2013; Dixon et al , 2015). …”

Section: Challenges and Suggested Recommendations For Future Biobankimentioning

confidence: 99%

Precision medicine in cancer: challenges and recommendations from an EU-funded cervical cancer biobanking study

et al. 2016

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Background:Cervical cancer (CC) remains a leading cause of gynaecological cancer-related mortality worldwide. CC pathogenesis is triggered when human papillomavirus (HPV) inserts into the genome, resulting in tumour suppressor gene inactivation and oncogene activation. Collecting tumour and blood samples is critical for identifying these genetic alterations.Methods:BIO-RAIDs is the first prospective molecular profiling clinical study to include a substantial biobanking effort that used uniform high-quality standards and control of samples. In this European Union (EU)-funded study, we identified the challenges that were impeding the effective implementation of such a systematic and comprehensive biobanking effort.Results:The challenges included a lack of uniform international legal and ethical standards, complexities in clinical and molecular data management, and difficulties in determining the best technical platforms and data analysis techniques. Some difficulties were encountered by all investigators, while others affected only certain institutions, regions, or countries.Conclusions:The results of the BIO-RAIDs programme highlight the need to facilitate and standardise regulatory procedures, and we feel that there is also a need for international working groups that make recommendations to regulatory bodies, governmental funding agencies, and academic institutions to achieve a proficient biobanking programme throughout EU countries. This represents the first step in precision medicine.

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“…The advent of tyramide signal amplification (TSA) improved the options for combining conspecific antibodies in the same sample while providing signal amplification if combined with an antibody stripping technique. (Buchwalow et al, 2018;Chao et al, 1996;Dixon et al, 2015;Lim et al, 2018;Mansfield, 2017;Pirici et al, 2009;Roy et al, 2019;Sorrelle et al, 2019;Stack et al, 2014;Wang et al, 1999;Zhang et al, 2017). In short, TSA uses a peroxidase-mediated reaction to covalently deposit fluorophores or chromogens to tyrosine side chains proximal to the target epitope (Lim et al, 2018;Wang et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Multiplex immunofluorescence methods in neurodegenerative diseases

Ehrenberg

Morales

Tejedor

et al. 2019

Preprint

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BACKGROUND: Neurodegenerative diseases feature stereotypical deposits of protein aggregates that selectively accumulate in vulnerable cells. The ability to simultaneously localize multiple targets in situ would facilitate discovery and validation of molecular pathways underlying neurodegenerative diseases.Immunostaining methods offer in situ detection of targets. Most proposed protocols for multiplexing immunostaining require sophisticated equipment not available for standard labs. Effective stripping of antibodies, allowing successive rounds of staining while maintaining tissue and antigen integrity, is the main roadblock for enabling multiplex immunostaining in standard labs. Antibody stripping techniques have not been fully validated for neurodegenerative disease research. Moreover, despite the increased complexity of multiplex immunostaining protocols, quality control steps have not been regularly implemented.NEW METHOD: Aiming to create protocols for multiplex localization of neurodegenerative-related processes, without the need for specialized equipment, we evaluated several antibody stripping techniques. We also recommend quality control steps to validate stripping efficacy and ameliorate concerns of cross-reactivity and false positives. RESULTS:The proposed protocol using β -mercaptoethanol enables reliable antibody stripping across multiple rounds of staining and minimizes the odds of cross-reactivity while preserving tissue and antigen integrity.COMPARISON WITH EXISTING METHODS: Our proposed method accounts for the intricacies of suboptimal human post-mortem tissue and the need to strip markers bound to highly aggregated proteins.Additionally, it incorporates quality control steps to validate antibody stripping.CONCLUSIONS: Multiplex immunofluorescence methods for studying neurodegenerative diseases in human postmortem tissue are feasible even in standard laboratories. Nevertheless, evaluation of stripping parameters in optimization and validation phases of experiments is prudent.

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Recent developments in multiplexing techniques for immunohistochemistry

Cited by 103 publications

References 87 publications

Detection approaches for multidrug resistance genes of leukemia

Detection approaches for multidrug resistance genes of leukemia

Precision medicine in cancer: challenges and recommendations from an EU-funded cervical cancer biobanking study

Multiplex immunofluorescence methods in neurodegenerative diseases

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