Slide 1 luer 0.8 channel slide

The Parsortix device was operated using the procedures as previously described [25 (link)]. MDA-MB-231TD (1 × 10³) cells were spiked into 7.5 mL of whole blood and the vacutainer was attached directly onto the Parsortix device. Cells were harvested after isolation and tethered onto a µ-Slide I Luer 0.8 Ibidi channel slide (Ibidi, Germany) (Cat: 80191) coated with PEM + DOTAP for 30 min. Live cells were stained with CellMask Orange (Thermo Fisher Scientific) (Cat: C10045) and NucBlue Live ReadyProbes Reagent (Thermo Fisher Scientific) (Cat: R37605) to visualize the plasma membrane and nucleus, respectively. Images were acquired using an Olympus IX81 microscope with a Fluoview FV1000 confocal laser scanning system.

The Vortex VTX-1 device was operated using the procedures as previously described [43 (link)]. MDA-MB-231TD cells (5 × 10³) were spiked into 4 mL of whole blood diluted in PBS to a total volume of 40 mL, for a 10X blood dilution. Cells were isolated with the VTX-1 and collected into a 1.5 mL eppendorf tube (~ 300 uL total volume in PBS).
Half of the collected sample (~ 150 uL) was treated with 0.1% DMSO and the other half of the sample was treated with 10 μM Vinorelbine for 1 h in a 96-well low-attach plate (Corning, Corning, NY) (Cat: 3474). Total volume was collected for both samples and tethered onto a µ-Slide I Luer 0.8 Ibidi channel slide (Ibidi, Germany) (Cat: 80191) coated with PEM + DOTAP for 30 min. Cells were then fixed with 3.7% formaldehyde and stained with Hoechst 33258 (1:5000) and Wheat Germ Agglutinin (WGA, Alexa Fluor 594) (1:100) (Thermo Fisher Invitrogen) (Cat: W11262) to visualize the nucleus and cell membrane, respectively. Images were acquired using an Olympus IX81 microscope with a Fluoview FV1000 confocal laser scanning system.