Increasing insulin measurement throughput by fluorescence anisotropy imaging immunoassays

Wang, Yao; Adeoye, Damilola I.; Wang, Yue J.; Roper, Michael G.

doi:10.1016/j.aca.2022.339942

Cited by 15 publications

(15 citation statements)

References 46 publications

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“…[6][7][8][9][10][11][12][13][14] To increase throughput while simplifying flow control, vacuum-based fluidic transport can be used. [15][16][17][18][19] Typically, a single vacuum line is applied to the outlet of the device and the resistances of each channel determine the flow rates. Active control from one or several channels in this type of system is more difficult to achieve.…”

Section: Introductionmentioning

confidence: 99%

“…For example, in the devices that have used vacuum-based transport for examining secretion from islets, no active control over the flow in any of the channels was reported. [16][17][18] This lack of control required manual replacement of solutions that changed composition over time, for example, perfusion solutions that contained different glucose concentrations to stimulate islets. In addition to being cumbersome, increased manual manipulation can lead to contamination of the device or reagents.…”

Section: Introductionmentioning

confidence: 99%

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Automated cellular stimulation with integrated pneumatic valves and fluidic capacitors

Adeoye

Wang

Davis

et al. 2023

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Automated cellular stimulation with integrated pneumatic valves and fluidic capacitors

Adeoye

Wang

Davis

et al. 2023

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“…Notably, existing analytical methods to measure secreted insulin employ microfluidic devices and (competing) immunoassays. For example, Roper and co-workers developed electrophoretic assays on microfluidic devices that measure insulin dynamics every 10 s, where insulin secreted from the islets competes with fluorescently labeled insulin for a limited amount of antibody. Our approach could complement such methods because IMS-MS can reliably distinguish between the two rodent insulin isoforms …”

Section: Resultsmentioning

confidence: 99%

Mouse Pancreatic Peptide Hormones Probed at the Sub-Single-Islet Level: The Effects of Acute Corticosterone Treatment

et al. 2022

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We combine liquid chromatography coupled with ion mobility spectrometry−mass spectrometry to elucidate how short exposure to corticosterone (Cort) alters the output of mouse pancreatic islet hormones. The workflow enables the robust separation of mouse insulin 1 (Ins1) and insulin 2 (Ins2) and the detection of major islet hormones in a homogenate equivalent to 100−150 islet cells. We show that Ins2 has a unique structure and is degraded much faster than Ins1. Further investigation indicates that Ins2 may populate both T and R states, whereas Ins1 may not. The assemblies of Ins1's B-chain also introduce more structural heterogeneity than Ins2. Collectively, these features account for their unique degradation profiles, the diabetes risk associated with Ins1, and the protective effect of Ins2. In the same experiments, we observe that the ratio of amylin to Ins1 increased significantly in Cort-treated mice (15:1) compared to the control mice (42:1), correlating well with β-cell proliferation observed in immunoassays on the same animal model. We observe no increase in intact full-length insulin levels but more of the truncated forms, indicating that enzymatic activity is accelerated. Our data provide a molecular basis for reduced insulin action induced by Cort and connections between insulin turnover and insulin resistance.

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“…Adablah, in the same group as Schrell, replaced the Cy5 with SeTAu-647 characterized by a longer lifetime and higher brightness, and the alteration resulted in a 45% increase in anisotropy range and a significant improvement in signal-to-noise ratio[ 54 ]. With the development of this technology, a highly integrated chip for separately sensing up to 12 islet secretomes was designed by Wang et al [ 55 ], this integration ensures the possibility of high-throughput single islet secretome monitoring, and the procedure for islet heterogeneity study is markedly simplified.…”

Section: Fluorescence Anisotropy Methodsmentioning

confidence: 99%

Advances in microfluidic chips based on islet hormone-sensing techniques

Li¹,

Peng²

2023

World J Diabetes

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Diabetes mellitus is a global health problem resulting from islet dysfunction or insulin resistance. The mechanisms of islet dysfunction are still under investigation. Islet hormone secretion is the main function of islets, and serves an important role in the homeostasis of blood glucose. Elucidating the detailed mechanism of islet hormone secretome distortion can provide clues for the treatment of diabetes. Therefore, it is crucial to develop accurate, real-time, labor-saving, high-throughput, automated, and cost-effective techniques for the sensing of islet secretome. Microfluidic chips, an elegant platform that combines biology, engineering, computer science, and biomaterials, have attracted tremendous interest from scientists in the field of diabetes worldwide. These tiny devices are miniatures of traditional experimental systems with more advantages of time-saving, reagent-minimization, automation, high-throughput, and online detection. These features of microfluidic chips meet the demands of islet secretome analysis and a variety of chips have been designed in the past 20 years. In this review, we present a brief introduction of microfluidic chips, and three microfluidic chips-based islet hormone sensing techniques. We focus mainly on the theory of these techniques, and provide detailed examples based on these theories with the hope of providing some insights into the design of future chips or whole detection systems.

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Increasing insulin measurement throughput by fluorescence anisotropy imaging immunoassays

Cited by 15 publications

References 46 publications

Automated cellular stimulation with integrated pneumatic valves and fluidic capacitors

Automated cellular stimulation with integrated pneumatic valves and fluidic capacitors

Mouse Pancreatic Peptide Hormones Probed at the Sub-Single-Islet Level: The Effects of Acute Corticosterone Treatment

Advances in microfluidic chips based on islet hormone-sensing techniques

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