Detection of red blood cell surface antigens by probe-triggered cell collision and flow retardation in an autonomous microfluidic system

Sautner, Éva; Papp, Krisztián; Holczer, Eszter; Tóth, Eszter L.; Ungai-Salánki, Rita; Szabó, Bálint; Fürjes, Péter; Prechl, József

doi:10.1038/s41598-017-01166-9

Cited by 6 publications

(5 citation statements)

References 14 publications

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“…Essentially, 1 mW of laser power approximately generates 1 pN force for a 1 μm diameter sphere [44]. The experiment subject, RBC, is 7.5~8.5 μm in diameter, with a width of 1.7~2.2 μm in the ring and 0.5~1 μm at the center [45]. In this study, the optical trapping spot exerts at a constant volume at the ring edge of an attached RBC during cell-tearing assays.…”

Section: The Force (F) Can Be Expressed Asmentioning

confidence: 94%

A novel methodology for detecting variations in RBCs surface antigens by cell-tearing using optical tweezers

Lin

Wang

Tien

et al. 2022

Preprint

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Background: The quantitative analysis of cell surface antigens has attracted increasing attention due to the antigenic variation recognition that can facilitate diagnoses. The whole-cell-based analysis minimizes sample size, suggesting an alternative approach for detecting the variation in cell surface antigens. The optical tweezers (OT) are practical for precise single-cell manipulations, and has been widely used as a convenient tool in interdisciplinary fields. However, employing this technology as a biosensor using a "tearing" operation is still rare. Method:This paper presents a novel methodology based on the “tearing” operation of optical tweezers (OT) incorporated with the “dilution method” of antibodies to detect variations in red blood cell (RBC) surface antigens. The RBCs attach to the corresponding antibody-coated cover glass. Then, the binding firmness between an RBC and the functionalized surface is assayed by optically tearing using gradually reduced laser powers incorporated with serial antibody dilutions. Results:The experiment result shows that the higher dilution (lower antibody concentration), the lower power (lower optical force) needed to tear off the RBC binding from the functionalized surface, i.e., the antibody dilution fold is inversely proportional to the laser power. With the relative-quantitative analysis, the variation in RBC surface antigens can be intuitively estimated by comparing the maximum allowable dilution folds. The estimation gives that the antigens on the B3-type RBC are 35.7% of that on the B-type RBC, which is consistent with the literature findings using conventional biological methods. Conclusions:This study proposes a new application of the optical tweezers as a biosensor using the optically cell-tearing operation to estimate the variation in RBC surface antigens. With the proposed methodology, the detection of antigenic variation was successfully implemented without complicated optical force calculations and labored biological processing. One drop of blood from the fingertip is more than enough to obtain a satisfactory detection. It suggests an novel approach for antigenic variation analyses based on the whole-cell operation.

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Section: The Force (F) Can Be Expressed Asmentioning

confidence: 94%

A novel methodology for detecting variations in RBCs surface antigens by cell-tearing using optical tweezers

Lin

Wang

Tien

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Essentially, 1 mW of laser power approximately generates 1 pN force for a 1 μm diameter sphere [ 50 ]. In this study, the experiment subject, RBC, is 7.5~8.5 μm in diameter, with a width of 1.7~2.2 μm in the ring and 0.5~1 μm at the center [ 51 ]. The optical trapping spot exerts at a constant volume at the ring edge of an attached RBC during cell-tearing assays.…”

Section: Methodsmentioning

confidence: 99%

A Novel Methodology for Detecting Variations in Cell Surface Antigens Using Cell-Tearing by Optical Tweezers

et al. 2022

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The quantitative analysis of cell surface antigens has attracted increasing attention due to the antigenic variation recognition that can facilitate early diagnoses. This paper presents a novel methodology based on the optical “cell-tearing” and the especially proposed “dilution regulations” to detect variations in cell surface antigens. The cell attaches to the corresponding antibody-coated slide surface. Then, the cell-binding firmness between a single cell and the functionalized surface is assayed by optically tearing using gradually reduced laser powers incorporated with serial antibody dilutions. Groups B and B3 of red blood cells (RBCs) were selected as the experiment subject. The results indicate that a higher dilution called for lower power to tear off the cell binding. According to the proposed relative-quantitative analysis theory, antigenic variation can be intuitively estimated by comparing the maximum allowable dilution folds. The estimation result shows good consistency with the finding in the literature. This study suggests a novel methodology for examining the variation in cell surface antigens, expected to be widely capable with potential sensor applications not only in biochemistry and biophysics, but also in the micro-/nano- engineering field.

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“…Essentially, 1 mW of laser power approximately generates 1 pN force for a 1 μm diameter sphere [40]. In this study, the experiment subject, RBC, is 7.5~8.5 μm in diameter, with a width of 1.7~2.2 μm in the ring and 0.5~1 μm at the center [41]. During cell-detachment testing, the optical trapping spot exerts at a constant volume at the ring edge of an attached RBC.…”

Section: Optical Tweezers For Rbc Manipulationmentioning

confidence: 97%

A simple and intuitive methodology for estimating red blood cell surface antigen expression variation using optical cell-detachment technique

Lin

Wang

TIEN³

et al. 2021

Preprint

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The analysis of surface antigens on cells, especially red blood cells (RBCs), has attracted increasing attention due to the recognition of antigenic variation that can facilitate early diagnoses. This paper presents an alternative methodology to estimate the variation of surface antigen expressions using an optical cell-detachment technique to validate the binding of individual RBCs stuck on corresponding antibody-coated surfaces. The detachment tests were implemented by an optical tweezers with gradually decreasing laser powers associated with serial antibody dilutions. Then, the antigen expression variation was estimated based on the known antibody dilution folds. The B- and B3-types of RBCs were selected for the demonstration subjects. With the semi-quantitative analysis, the proposed methodology was successfully verified for evaluating the variation of the RBC surface antigen expressions. The analysis result shows good consistency with the literature’s findings.

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Detection of red blood cell surface antigens by probe-triggered cell collision and flow retardation in an autonomous microfluidic system

Cited by 6 publications

References 14 publications

A novel methodology for detecting variations in RBCs surface antigens by cell-tearing using optical tweezers

A novel methodology for detecting variations in RBCs surface antigens by cell-tearing using optical tweezers

A Novel Methodology for Detecting Variations in Cell Surface Antigens Using Cell-Tearing by Optical Tweezers

A simple and intuitive methodology for estimating red blood cell surface antigen expression variation using optical cell-detachment technique

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