Quantitative proteomic profiling of breast cancers using a multiplexed microfluidic platform for immunohistochemistry and immunocytochemistry

Kim, Minseok S.; Kwon, Soon-Young; Kim, Taemin; Lee, Eun Sook; Park, Je‐Kyun

doi:10.1016/j.biomaterials.2010.10.040

Cited by 28 publications

(29 citation statements)

References 32 publications

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“…The precision of microfluidic devices has improved existing single-cell assays (such as immunocytochemistry 23 and immunohistochemistry 24,25 ) and has bolstered the invention of novel assays for quantitative analysis of thousands of cells with single-cell resolution 26 .…”

Section: Single-cell Measurementsmentioning

confidence: 99%

Microfluidics: reframing biological enquiry

Duncombe

Tentori

Herr

2015

Nat Rev Mol Cell Biol

294

202

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The underlying physical properties of microfluidic tools have led to new biological insights through the development of microsystems that can manipulate, mimic and measure biology at a resolution that has not been possible with macroscale tools. Microsystems readily handle sub-microlitre volumes, precisely route predictable laminar fluid flows and match both perturbations and measurements to the length scales and timescales of biological systems. The advent of fabrication techniques that do not require highly specialized engineering facilities is fuelling the broad dissemination of microfluidic systems and their adaptation to specific biological questions. We describe how our understanding of molecular and cell biology is being and will continue to be advanced by precision microfluidic approaches and posit that microfluidic tools — in conjunction with advanced imaging, bioinformatics and molecular biology approaches — will transform biology into a precision science.

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Section: Single-cell Measurementsmentioning

confidence: 99%

Microfluidics: reframing biological enquiry

Duncombe

Tentori

Herr

2015

Nat Rev Mol Cell Biol

294

202

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“…In addition to the 10-fold decrease in assay time and reagent consumption, automated microfluidic-mediated labeling permits control over the reagent flow rate, which can be optimized to increase receptor-ligand binding. This microfluidic array design has been scaled up to permit analysis of 10 biomarkers (Figure 3B1) in a single experiment [47]. This group also compared an 8-array microfluidic staining method, to label four breast cancer markers with quantum dots, to conventional multiplexing methods that were either sequential, using secondary antibodies from the same species for each cycle, or simultaneous, using a cocktail of antibodies from dissimilar species in single cycle.…”

Section: Spatially-patterned Multiplexing For Multiple Tissue Regionsmentioning

confidence: 99%

“…Reproduced with permission from [47], Biomaterials, 32(5), Kim M, Kwon S, Kim T, Lee E, Park J-K, Quantitative proteomic profiling of breast cancers using a multiplexed microfluidic platform for immunohistochemistry and immunocytochemistry, 1396–1403, Copyright 2011, with permission from Elsevier [47]. …”

Section: Figurementioning

confidence: 99%

“…All figures reproduced with permissions as follows: permission obtained from Nature Publishing Group [30], PLOS One [46], Biomaterials,32(5), Kim M, Kwon S, Kim T, Lee E, Park J-K. Quantitative proteomic profiling of breast cancers using a multiplexed microfluidic platform for immunohistochemistry (IHC) and immunocytochemistry, 1396–1403, Copyright 2011, with permission from Elsevier [47]. Reprinted with Permission of SAGE Publications [50,51], and Reprinted with permission from John Wiley and Sons [60].…”

Section: Figurementioning

confidence: 99%

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Recent developments in multiplexing techniques for immunohistochemistry

Dixon

Bathany

Tsuei

et al. 2015

Expert Review of Molecular Diagnostics

103

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Methods to detect immuno-labelled molecules at increasingly higher resolution, even when present at low levels, are revolutionizing immunohistochemistry (IHC). These technologies can be valuable for management and examination of rare patient tissue specimens, and for improved accuracy of early disease detection. The purpose of this mini-review is to highlight recent multiplexing methods that are candidates for more prevalent use in clinical research and potential translation to the clinic. Multiplex IHC methods, which permit identification of at least 3 and up to 30 discrete antigens, have been divided into whole section staining and spatially-patterned staining categories. Associated signal enhancement technologies that can enhance performance and throughput of multiplex IHC assays are also discussed. Each multiplex IHC technique, detailed herein, is associated with several advantages as well as tradeoffs that must be taken into consideration for proper evaluation and use of the methods.

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“…The system could work mostly in the diffusion-limited mode, and the signal was detected using a special sophisticated time-resolved fluorescent detection method. Other researchers opted for multiplex IHC (16,17) with multiple parallel small channels targeting different biomarkers at spatially displaced locations using higher-antibody dilutions than needed for conventional IHC analysis. However, none of these microfluidic approaches resulted in an increase in accuracy of quantitative biomarker expression analysis and hence a reduction of ambiguous diagnostic results.…”

mentioning

confidence: 99%

Microfluidic processor allows rapid HER2 immunohistochemistry of breast carcinomas and significantly reduces ambiguous (2+) read-outs

Çiftlik

Lehr

Gijs

2013

Proc. Natl. Acad. Sci. U.S.A.

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Biomarker analysis is playing an essential role in cancer diagnosis, prognosis, and prediction. Quantitative assessment of immunohistochemical biomarker expression on tumor tissues is of clinical relevance when deciding targeted treatments for cancer patients. Here, we report a microfluidic tissue processor that permits accurate quantification of the expression of biomarkers on tissue sections, enabled by the ultra-rapid and uniform fluidic exchange of the device. An important clinical biomarker for invasive breast cancer is human epidermal growth factor receptor 2 [(HER2), also known as neu], a transmembrane tyrosine kinase that connotes adverse prognostic information for the patients concerned and serves as a target for personalized treatment using the humanized antibody trastuzumab. Unfortunately, when using state-of-the-art methods, the intensity of an immunohistochemical signal is not proportional to the extent of biomarker expression, causing ambiguous outcomes. Using our device, we performed tests on 76 invasive breast carcinoma cases expressing various levels of HER2. We eliminated more than 90% of the ambiguous results (n = 27), correctly assigning cases to the amplification status as assessed by in situ hybridization controls, whereas the concordance for HER2-negative (n = 31) and -positive (n = 18) cases was 100%. Our results demonstrate the clinical potential of microfluidics for accurate biomarker expression analysis. We anticipate our technique will be a diagnostic tool that will provide better and more reliable data, onto which future treatment regimes can be based.

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Quantitative proteomic profiling of breast cancers using a multiplexed microfluidic platform for immunohistochemistry and immunocytochemistry

Cited by 28 publications

References 32 publications

Microfluidics: reframing biological enquiry

Microfluidics: reframing biological enquiry

Recent developments in multiplexing techniques for immunohistochemistry

Microfluidic processor allows rapid HER2 immunohistochemistry of breast carcinomas and significantly reduces ambiguous (2+) read-outs

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