Automated Microscopic Analysis of Thrombus Formation

From each microspot, two bright-field images (during labeling) and three 3-colour fluorescence images (after removing the label) were taken using an EVOS-FL microscope (Life Technologies, Bleiswijk, the Netherlands) equipped with Cy5, RFP, and GFP LEDs; an Olympus UPLSAPO 60× oil immersion objective; and a sensitive 1360 × 1024 pixel CCD camera [49 (link)]. A standardized image analysis was performed using semiautomated scripts operated in Fiji (ImageJ), as described before [49 (link)]. Parameters extracted from bright-field images (P1–5), including thrombus signature scores (P3–5), and parameters from fluorescence images (P6–8) are specified in Table 1.

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Jooss N.J., De Simone I., Provenzale I., Fernández D.I., Brouns S.L., Farndale R.W., Henskens Y.M., Kuijpers M.J., ten Cate H., van der Meijden P.E., Cavill R, & Heemskerk J.W. (2019). Role of Platelet Glycoprotein VI and Tyrosine Kinase Syk in Thrombus Formation on Collagen-Like Surfaces. International Journal of Molecular Sciences, 20(11), 2788.

Publication 2019

EVOS-FL microscope UPLSAPO 60×

Fluorescence Immersion Microscope Thrombus

Corresponding Organization : Maastricht University

Other organizations : University of Cambridge

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Variable analysis

independent variables

The protocol does not explicitly mention any independent variables that were manipulated by the researchers.

dependent variables

Parameters extracted from bright-field images (P1–5), including thrombus signature scores (P3–5)
Parameters from fluorescence images (P6–8)

control variables

The protocol does not explicitly mention any control variables that were kept constant to ensure valid results.

controls

The protocol does not specify any positive or negative controls.

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