Flat clear bottom black 96 well plates

Pre-treated SPC-548-OVA expressing tumour cells (GFP positive) were plated in triplicate in flat clear bottom black 96-well plates (Corning) coated with 5 μg/mL of fibronectin (Merck) prior to plating. T cells were added at 2:1 (effector:target) ratio. Plates were incubated for 24 hours in the Incucyte SX5 Live Cells Analysis System at 37°C. Images were acquired every 4 hours measuring green object count (GFP positive cells). Depletion of OVA expressing tumour cells in the presence and absence of T cells was calculated by fold-change of green object count at each time point normalised to T0.

Single-stranded 5′-GTG TGT GTG TGT-3′ [(GT)₆] DNA oligonucleotides were purchased from Integrated DNA technologies (standard desalting and lyophilized powder), and HiPco single wall carbon nanotubes were purchased from Nano Integris (batch #HR35-141). Solution-phase nanosensor synthesis was carried out by first mixing 1 mg of (GT)₆ and 1 mg of HiPco SWNT in 1 mL of 1 × PBS. Then the solution was bath-sonicated (Branson 1800) at room temperature for 20 min followed by probe-tip sonication (Sonics Vibra Cell) for 15 min in an ice-bath. Resulting suspension was centrifuged at 20,000 rcf (Eppendorf 5430 R) at 4°C for 60 min, and supernatant was carefully transferred into a new Eppendorf tube and stored at 4°C until further use. For characterization, each sensor batch was diluted to a working concentration of 10 ppm in 1 × PBS. Fluorescence and absorbance measurements were carried out on NS Super NanoSpectralyzer (Applied Nanotechnologies). Solution-phase fluorescence measurements and dopamine response tests were carried out on a custom built near infrared plate reader using flat clear bottom black 96-well plates (Corning, #3904). Solution-phase ∆F/F values were calculated from integrals of fluorescence counts between 875 and 1300 nm for pre- and post-dopamine fluorescence spectra.