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TECHNICAL PAPERS

A Modified Micropipette Aspiration Technique and Its Application to Tether Formation From Human Neutrophils

[+] Author and Article Information
Jin-Yu Shao, Jinbin Xu

Department of Biomedical Engineering, Washington University, Saint Louis, MO 63130

J Biomech Eng 124(4), 388-396 (Jul 30, 2002) (9 pages) doi:10.1115/1.1486469 History: Received September 01, 2001; Revised April 01, 2002; Online July 30, 2002
Copyright © 2002 by ASME
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References

Figures

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A diagram of the modified micropipette manipulation system. Another micropipette can be set up on the other side of the chamber if two micropipettes are needed.
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A video micrograph showing tether formation with two micropipettes. The cell shown here is a human neutrophil stimulated with 50 ng/ml PMA.
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Two video micrographs showing how to measure the residual cortical tension of a neutrophil treated with 100 μM cytochalasin D by aspirating the non-blebby region of the same cell with two micropipettes whose diameters are 5.2 μm and 3.3 μm respectively
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The displacement of a latex bead fixed on a glass cover slip over time. The displacement was tracked with BeadPro8 and BeadPro18 respectively at a speed of 30 frames/second. Shown by the legends are the average bead positions and the standard deviations.
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The displacement of a latex bead moving towards and away from the opening of a micropipette under an alternating net pressure of 1 pN/μm2 . Here, the velocities of the bead are ∼4 μm/s.
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Tether formation from passive neutrophils with the neutrophil as the transducer (circle) and the bead as the transducer (cross). The solid line represents a linear fit through the circles and the dashed line represents a linear fit through the crosses. Each cross represents an average of two to eight tethers from the same cell at a certain pressure. The error bars stand for the standard deviations.
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Tether formation from neutrophils stimulated with 25 ng/ml IL-8. The solid line represents Eq. (5) and each cross represents an average of three to seven tethers from the same cell at a certain pressure. The error bars stand for the standard deviations.
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Tether formation from neutrophils stimulated with 50 ng/ml PMA. The solid line represents Eq. (5) and each cross represents an average of five to ten tethers from the same cell at a certain pressure. The error bars stand for the standard deviations.
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Tether formation from neutrophils treated with 100 μM cytochalasin D. The solid line represents Eq. (5) and each cross represents an average of one to seventeen tethers from the same cell at a certain pressure. The error bars stand for the standard deviations.
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A doublet of two spherical beads on a flat surface. OA and OB are the centers of the two beads. A and B are the projections of OA and OB on the flat surface respectively. P is the projection of OA on OBB.

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