Development of single-particle counting assays with interferometric reflectance imaging

Biomarkers are biological measures used for clinical assessment, whether an individual has a particular medical condition or to monitor and predict health states in individuals. Sensitive detection and quantification of various biomarkers are essential for disease diagnostics. The majority of biomar...

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Main Author: Ekiz Kanik, Fulya
Other Authors: Ünlü, Selim M.
Language:en_US
Published: 2020
Subjects:
Online Access:https://hdl.handle.net/2144/41477
id ndltd-bu.edu-oai-open.bu.edu-2144-41477
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spelling ndltd-bu.edu-oai-open.bu.edu-2144-414772020-10-18T17:01:26Z Development of single-particle counting assays with interferometric reflectance imaging Ekiz Kanik, Fulya Ünlü, Selim M. Electrical engineering Assay development Biosensor Interferometry Nucleic acid detection Protein detection Virus detection Biomarkers are biological measures used for clinical assessment, whether an individual has a particular medical condition or to monitor and predict health states in individuals. Sensitive detection and quantification of various biomarkers are essential for disease diagnostics. The majority of biomarker-based diagnostics examines the presence and quantity of a single biomarker. Since the symptoms of many diseases are alike, multiplexed biomarker tests are highly desirable. Furthermore, detection of multiple biomarkers would improve the accuracy of diagnosis as well as providing additional information about the prognosis. Microarray platforms have the potential for higher level of multiplexing for biomarker detection. However, conventional microarray technologies are limited by the sensitivity of assays. This dissertation describes how single-particle interferometric reflectance imaging sensor (SP-IRIS) overcomes the sensitivity issues in biomarker detection and its applications to biomolecular and cellular biomarker detection assays. SP-IRIS provides optical detection of individual nanoparticles when they are captured onto a simple reflecting substrate, providing single-molecule sensitivity. This technique can be used to detect natural nanoparticles (such as viruses) without labels as well as molecular analytes (proteins and nucleic acids) that are labeled with metallic nanoparticles. Moreover, the advancements in technology make SP-IRIS ideal for the detection of low abundance biomarkers. Utilization of light polarization in combination with plasmonic gold nanorods as labels enhances the signal-to-noise ratio in nanoparticle detection allowing for the use of low numerical aperture optics increasing the field-of-view, hence, the throughput and sensitivity. Additionally, the integration of a disposable microfluidic flow cell and dynamic particle tracking in kinetic measurements provide a robust, ultra-sensitive and automated diagnostic platform. This dissertation focuses on the development of biological assays demonstrating effective use of SP-IRIS as a clinical diagnostic platform. We discuss the development of protein, nucleic acid and biological nanoparticle detecting SP-IRIS microarrays. We demonstrate four digital detection platforms for Hepatitis B, microRNA, rare mutations in an oncogene, KRAS, and virus-like particle detection with ultra-high sensitivity. 2022-09-28T00:00:00Z 2020-10-16T19:07:59Z 2020 2020-09-29T04:02:03Z Thesis/Dissertation https://hdl.handle.net/2144/41477 0000-0003-2662-7832 en_US Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/
collection NDLTD
language en_US
sources NDLTD
topic Electrical engineering
Assay development
Biosensor
Interferometry
Nucleic acid detection
Protein detection
Virus detection
spellingShingle Electrical engineering
Assay development
Biosensor
Interferometry
Nucleic acid detection
Protein detection
Virus detection
Ekiz Kanik, Fulya
Development of single-particle counting assays with interferometric reflectance imaging
description Biomarkers are biological measures used for clinical assessment, whether an individual has a particular medical condition or to monitor and predict health states in individuals. Sensitive detection and quantification of various biomarkers are essential for disease diagnostics. The majority of biomarker-based diagnostics examines the presence and quantity of a single biomarker. Since the symptoms of many diseases are alike, multiplexed biomarker tests are highly desirable. Furthermore, detection of multiple biomarkers would improve the accuracy of diagnosis as well as providing additional information about the prognosis. Microarray platforms have the potential for higher level of multiplexing for biomarker detection. However, conventional microarray technologies are limited by the sensitivity of assays. This dissertation describes how single-particle interferometric reflectance imaging sensor (SP-IRIS) overcomes the sensitivity issues in biomarker detection and its applications to biomolecular and cellular biomarker detection assays. SP-IRIS provides optical detection of individual nanoparticles when they are captured onto a simple reflecting substrate, providing single-molecule sensitivity. This technique can be used to detect natural nanoparticles (such as viruses) without labels as well as molecular analytes (proteins and nucleic acids) that are labeled with metallic nanoparticles. Moreover, the advancements in technology make SP-IRIS ideal for the detection of low abundance biomarkers. Utilization of light polarization in combination with plasmonic gold nanorods as labels enhances the signal-to-noise ratio in nanoparticle detection allowing for the use of low numerical aperture optics increasing the field-of-view, hence, the throughput and sensitivity. Additionally, the integration of a disposable microfluidic flow cell and dynamic particle tracking in kinetic measurements provide a robust, ultra-sensitive and automated diagnostic platform. This dissertation focuses on the development of biological assays demonstrating effective use of SP-IRIS as a clinical diagnostic platform. We discuss the development of protein, nucleic acid and biological nanoparticle detecting SP-IRIS microarrays. We demonstrate four digital detection platforms for Hepatitis B, microRNA, rare mutations in an oncogene, KRAS, and virus-like particle detection with ultra-high sensitivity. === 2022-09-28T00:00:00Z
author2 Ünlü, Selim M.
author_facet Ünlü, Selim M.
Ekiz Kanik, Fulya
author Ekiz Kanik, Fulya
author_sort Ekiz Kanik, Fulya
title Development of single-particle counting assays with interferometric reflectance imaging
title_short Development of single-particle counting assays with interferometric reflectance imaging
title_full Development of single-particle counting assays with interferometric reflectance imaging
title_fullStr Development of single-particle counting assays with interferometric reflectance imaging
title_full_unstemmed Development of single-particle counting assays with interferometric reflectance imaging
title_sort development of single-particle counting assays with interferometric reflectance imaging
publishDate 2020
url https://hdl.handle.net/2144/41477
work_keys_str_mv AT ekizkanikfulya developmentofsingleparticlecountingassayswithinterferometricreflectanceimaging
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