Cfi plan apochromat lambda 100 oil

We calibrated the performance of the system using 0.2 µm fluorescent microspheres (TetraSpeck^TM, ThermoFisher Scientific, UK). The microspheres were excited by scanning an 8 × 8 beamlet array at 435 nm through a 100× NA1.45 objective (CFI Plan Apochromat Lambda 100× Oil, Nikon, UK) on an inverted microscope (Nikon Ti Eclipse, Nikon, UK) equipped with a perfect focus system. The effective pixel size was 62.5 nm in the x-y plane. The lateral and axial resolution was determined to be 302 ± 29 nm (s.d.) and 600 ± 96 nm (s.d.), respectively, from Gaussian fits to the data (Supplementary Fig. 1). Photon arrival times at the MF32 detector were calculated relative to a trigger (sync) signal from laser pulses incident on an optical constant fraction discriminator (OCF-401, Becker & Hickl, GmbH)) at the repetition rate of the laser, 80 MHz, which is sent to the MF32 detector. The temporal characterisation of the system was evaluated by adding a fixed delay of 5 ns to the photon arrival times by decreasing the sync cable (RG-174 type) length by 1 m. From measurements of the peak position and the decay curve from the sodium fluorescein solution we calculated the temporal bin size to be 57 ps. Representative intensity images of cells expressing empty vector mTurq2 and mTurq2-Epac1-td^dVenus and fluorescence decays from 5 × 5 pixel regions of the images are shown in Supplementary Fig. 2.

The CQD submonolayer films were made by spin-coating the CQD in n-octane dispersions on a cover slide. An objective lens (Nikon CFI Plan Apochromat Lambda 100× oil, numerical aperture of 1.45) was used to collect real-space and k-space emission patterns of the CQD films. To image k-space (BFP), two coupled 200-mm focal plano-convex lenses were placed on the back optical path of the objective lens and projected the BFP image of the objective lens onto a sCMOS (scientific complementary metal-oxide semiconductor) camera (TUCSEN Dhyana 400BSI V2). A polarizer (Thorlabs CCM1-PBS251) was used to generate the polarization image of the BFP, and a band-pass filter (Semrock FF02-641/75-25) was used to filter out the excitation and noise light. See text S2 for the BFP image fitting details.

The high-resolution Fourier light-field microscopy system (Fig. 1a and Supplementary Fig. 1) was developed using an epi-fluorescence microscope (Eclipse Ti2-U, Nikon Instruments)^{43 (link)}. The employed objective lens was an oil-immersion lens featuring 100× magnification and a numerical aperture (NA) of 1.45 (CFI Plan Apochromat Lambda 100× Oil, Nikon Instruments). A piezo nano-positioner (Nano-F100S, Mad City Labs) was utilized for precise positioning. Samples were excited using multicolor laser lines (488 nm, 561 nm, 647 nm, MPB Communications), with the fluorescence collected through a quadband dichroic mirror (ZT405/488/561/647, Chroma) and a corresponding emission filter (ZET405/488/561/647 m, Chroma). The sample stage incorporated a micro-positioning system (MS2000, Applied Scientific Instrumentation) for accurate placement. The native image plane of the objective lens was Fourier-transformed using a Fourier lens (f_FL = 275 mm, Edmund Optics). A customized microlens array (f_ML = 117 mm, RPC Photonics) was placed on the back focal plane of the Fourier lens (Supplementary Note 1). The elemental images formed by each microlens were captured using an sCMOS camera (ORCA-Flash 4.0 V3, Hamamatsu Photonics, pixel size P_cam = 6.5 µm).