Confocal laser-induced fluorescence detector for narrow capillary system with yoctomole limit of detection

Weaver, Mitchell T.; Lynch, Kyle B.; Zhu, Zaifang; Huang, Chen; Lü, Jianmin; Pu, Qiaosheng; Liu, Shaorong

doi:10.1016/j.talanta.2016.12.056

Cited by 41 publications

(20 citation statements)

References 19 publications

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“…For protein analysis method establishment, the performance of the CE-LIF system was evaluated first. The CE and LIF 25 The LOD level was much better than references reported recently 30 as well as commercial LIFs. In quantification of low copy number proteins in single cells, the cells were labeled by antibody, and then the single cell was introduced into the capillary by the siphon-injection.…”

mentioning

confidence: 65%

“…However, many of the latest analytical methods could not reach this sensitivity, even including various LIF applications. 30,37 Fortunately, our simple method could achieve an LOD of 135 molecules in the injection volume, which met the sensitivity demand of low copy number protein detection. In conclusion, we had established an ultrasensitive and simple CE-LIF method to quantify the low copy number of intracellular proteins at the single cell level.…”

mentioning

confidence: 96%

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Quantification of Low Copy Number Proteins in Single Cells

et al. 2019

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We have developed an ultrasensitive and highly selective method to quantify low copy number intracellular proteins in a single cell using a low-cost laser-induced fluorescence (LIF) detector and a BV605 fluorescent probe. Active caspase3 proteins in cells were labeled by corresponding antibody-BV605 fluorescent binding, and a cell was injected into a 20 cm × 50 μm i.d. capillary column, followed by in situ lysis and capillary electrophoresis (CE)-LIF analysis. About seven active caspase3 protein molecules in a detection volume of 91 pL could be detected. In our method, cross-bounding proteins other than active caspase3 could be separated and distinguished by differences of retention time. By using Si photodiode assembly as a fluorescent detector instead of PMT, the dynamic range of the LIF is over 4 orders of magnitude. In this experiment, we found that the number of active caspase3 molecules in 98 single Jurkat cells were from 629 to 12171, reflecting significant heterogeneity among the cells although they were from the same batch. For extended application, it could also be applied to quantify other types of low copy number proteins in a single cell as long as the corresponding antibodies are provided. This high-sensitive method could also be a promising tool for earlier cancer diagnosis and related disease pathway research which is relevant to low copy number proteins. In addition, this low-cost system could also be easily expanded to an array system for high-throughput quantitation of low copy proteins in single cells.

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confidence: 65%

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confidence: 96%

Quantification of Low Copy Number Proteins in Single Cells

et al. 2019

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“…Due to the small diameter of the separation column, the on-column LIF detector for BaNC-HDC is not commercially available. Weaver et al [ 58 ] describe the details of the construction method of a confocal LIF detector as well as how to adjust and use it for BaNC-HDC. The LIF detector was constructed using a vertical support which was firmly attached to a baseplate.…”

Section: Extension Of Hdc To Dna Separation: Experimentalmentioning

confidence: 99%

“… 1 kb plus DNA ladder BaNC-HDC This work described how to construct and adjust the confocal LIF for BaNC-HDC. [ 58 ] 2-μm i.d. 1 kb plus DNA ladder, plasmid DNA from E. coli BaNC-HDC An EOP and a microchip injector were incorporated with BaNC-HDC in this work.…”

Section: Introductionmentioning

confidence: 99%

Extension of hydrodynamic chromatography to DNA fragment sizing and quantitation

Wang

Zhou

Zhang

et al. 2021

Heliyon

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Hydrodynamic chromatography (HDC) is a technique originally developed for separating particles. We have recently extended it to DNA fragment sizing and quantitation. In this review, we focus on this extension. After we briefly introduce the history of HDC, we present the evolution of open tubular HDC for DNA fragment sizing. We cover both the theoretical aspect and the experimental implementation of this technique. We describe various approaches to execute the separation, discuss its representative applications and provide a future perspective of this technique in the conclusion section of this review.

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“…Since the introduction of IACE in the early 1990s by our laboratory, Terry Phillips’ laboratory, and others, the technology has increased significantly in popularity, and many applications have been reported using both conventional CE and microchip CE [ 25 , 54 , 55 , 58 , 61 , 62 , 63 , 64 , 66 , 75 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 ]. Furthermore, if IACE is coupled to powerful detectors such as laser-induced fluorescence detectors or mass spectrometers, a significant improvement in limits of detection can been achieved [ 25 , 75 , 99 , 100 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 ].…”

Section: Role Of Capillary Electrophoresis In Point-of-care Diagnomentioning

confidence: 99%

A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy

Guzman¹,

Guzman

2020

Biomedicines

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Biomarker detection for disease diagnosis, prognosis, and therapeutic response is becoming increasingly reliable and accessible. Particularly, the identification of circulating cell-free chemical and biochemical substances, cellular and subcellular entities, and extracellular vesicles has demonstrated promising applications in understanding the physiologic and pathologic conditions of an individual. Traditionally, tissue biopsy has been the gold standard for the diagnosis of many diseases, especially cancer. More recently, liquid biopsy for biomarker detection has emerged as a non-invasive or minimally invasive and less costly method for diagnosis of both cancerous and non-cancerous diseases, while also offering information on the progression or improvement of disease. Unfortunately, the standardization of analytical methods to isolate and quantify circulating cells and extracellular vesicles, as well as their extracted biochemical constituents, is still cumbersome, time-consuming, and expensive. To address these limitations, we have developed a prototype of a portable, miniaturized instrument that uses immunoaffinity capillary electrophoresis (IACE) to isolate, concentrate, and analyze cell-free biomarkers and/or tissue or cell extracts present in biological fluids. Isolation and concentration of analytes is accomplished through binding to one or more biorecognition affinity ligands immobilized to a solid support, while separation and analysis are achieved by high-resolution capillary electrophoresis (CE) coupled to one or more detectors. When compared to other existing methods, the process of this affinity capture, enrichment, release, and separation of one or a panel of biomarkers can be carried out on-line with the advantages of being rapid, automated, and cost-effective. Additionally, it has the potential to demonstrate high analytical sensitivity, specificity, and selectivity. As the potential of liquid biopsy grows, so too does the demand for technical advances. In this review, we therefore discuss applications and limitations of liquid biopsy and hope to introduce the idea that our affinity capture-separation device could be used as a form of point-of-care (POC) diagnostic technology to isolate, concentrate, and analyze circulating cells, extracellular vesicles, and viruses.

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Confocal laser-induced fluorescence detector for narrow capillary system with yoctomole limit of detection

Cited by 41 publications

References 19 publications

Quantification of Low Copy Number Proteins in Single Cells

Quantification of Low Copy Number Proteins in Single Cells

Extension of hydrodynamic chromatography to DNA fragment sizing and quantitation

A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy

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