Dp74 colour camera

The formation of liquid droplets was imaged on 50 μl of the N protein samples by DIC microscopy (OLYMPUS IX73 Inverted Microscope System with OLYMPUS DP74 Colour Camera) as previously described (Kang et al.,2019 (link)
a; Dang et al.,2021 ). The N protein samples were prepared at 10 μM in 25 mM HEPES buffer (pH 7.0) with 70 mM KCl (buffer 1), while NTD and CTD samples were prepared at 100 and 200 μM, respectively in 10 mM sodium phosphate buffer (pH 7.0) in the presence of 150 mM NaCl (buffer 2) for NMR studies. HCQ at 10 mM was dissolved in buffer 1 for LLPS or buffer 2 for NMR binding studies with the final pH adjusted to 7.0.

Before inoculation with worms, small amounts (approximately 0.5 g) of DFA medium were autoclaved in a glass beaker and transferred onto the centre of an NGM plate on which E. coli OP50 was seeded. Then, starved worms collected from two NGM plates were inoculated onto the DFA on the NGM plate. Worms propagated at 23 °C climbed up to the lid of the plate for approximately 10 days.
On the day of the experiment, a Petri plate was placed onto an aluminium plate on the stage of an Olympus SZX7 microscope, which was kept at 23 °C by a Peltier temperature controller unit (Vics). The Petri plate was maintained under this condition for 5 min before the image acquisition. Then, the temperature of the bottom of the Petri plate was increased from 23 to 26 °C to change the humidity inside the plate. Image acquisition of the inner surface of the plate lid was performed by the DP74 colour camera (Olympus) at 1.0 frame s⁻¹. A Plan Achromat objective lens (×0.5, NA = 0.05; Olympus) was used as a low-magnification objective. The acquired images were saved in the Tagged Image File format.