Wga alexa fluor 633 conjugate

A fibronectin (Sigma, USA) solution at a concentration of 100 μg ml⁻¹ was used to functionalize the glass surface of the microfluidic channel and to facilitate cell adhesion to the substrate. The cells were then seeded into the microfluidic channel and were incubated at 37 °C for at least two days before the experiments were conducted. The cell culture medium used for culturing MDA-MB-231 cells is composed of RPMI 1640 (Wisent Bioproducts, Canada) supplemented with 10% (v/v) fetal bovine serum (Wisent Bioproducts, Canada) and 1% (v/v) PenStrep (Wisent Bioproducts, Canada). For the enzymatic detachment experiments, MDA-MB-231 cells were incubated with 0.25% trypsin–EDTA (Wisent Bioproducts, Canada) for 4 min. To enzymatically dissociate HDMECs, we incubated them with trypsin and accutase (Gibco, Thermofisher, USA) for 15 and 10 min, respectively. We used the endothelium growth medium (PromoCell, Germany) for culturing HDMECs. For the cellular membrane staining, WGA Alexa Fluor 633 Conjugate (Thermofisher, USA) was used at a dilution ratio of 1 : 100 in the cell culture medium, and nuclear staining was carried out with Hoechst 33342 (Thermofisher, USA) at a concentration of 0.1 μg ml⁻¹. For the PI staining viability test, PI at a final concentration of 0.02 mg ml⁻¹ was used. The cells were incubated at 37 °C for 15 min before fluorescence images were taken.