Spinsr10 microscope

For images in Figure 1 and Videos 1–6:
HeLa cells were plated on 35-mm glass-bottom dishes (NEST, 801001) at a density of 0.13 × 10⁶. The indicated constructs were transfected into HeLa cells. Twelve to twenty-four hours after transfection, cells were imaged using a spinning-disk confocal microscope with a ×60 objective (Nikon T2 Microscope; Apo ×60 oil; 1.4 NA) using the SoRa mode. Exposure time was 500 ms. For time-lapse imaging, the time interval was set to 1 or 2 s. Images were analyzed with FIJI (ImageJ).
For images shown in supplements:
HeLa cells were plated on 35-mm glass-bottom dishes (BGI, BGX-03520-100) at a cell number of 0.8 × 10⁶ to grow for 12 hr. The indicated constructs were transfected into HeLa cells. Twelve hours after transfection, cells were imaged by a spinning-disk confocal microscope with a ×60 objective (Olympus SpinSR10 Microscope; Apo ×60 oil; 1.5 NA) using the SoRa mode. Different parameters were used:
Images shown in supplements: Exposure time was 500ms.
Time-lapse imaging for calculating the speed of mRNA movement: Exposure time was 200 ms, and the interval was set to 217 ms. Fifteen frames are recorded.
Z-stack imaging for calculating intensity and signal-to-noise: Exposure time was 500 ms. Stacks of 5 planes with a z-spacing of 0.5 μm were obtained by using range mode.

One-week-old Arabidopsis thaliana, WT (Col), and pldα1/δ knockout mutants were incubated with Nitrobenzoxadiazole (NBD) labeled phospholipids; NBD-PC or NBD-PA on a Corning glass slide (cat#2948) and covered with Corning coverslip (cat# 2980). Images were taken on an Olympus SpinSR10 microscope using a 40x/1.4 oil objective lens. Photos were taken in a z-stack of 30 µm thickness with a z-step size of 1µm. Fluorescence was detected using a 464 nm excitation and 531 nm emission. Obtained images were then projected on the z-axis using ImageJ (Fiji) software.