Health research capacity strengthening (HRCS) in the global South has become a major development objective, backed by arguments for the importance of national health research capacity to support locally-relevant, evidence-based health policy and practice [ 1– 5].
Wilson nonetheless carried his main objective back to America, a treaty for a "League of Nations," intended to prevent future wars.
In one-photon excitation, the objective back-pupil should be slightly under-filled with a parallel laser beam [19].
Illumination was 7 mW at the objective back aperture, at 810 nm for the FRET probes and at 910 nm for EGFP and the GCaMP2 probes.
Alternatively, polarized emission from the dipole converts to a spatial representation at the objective back focal plane [7], [8] that the tube lens images as the PSF at the camera.
An excitation filter (D460/50x, Chroma) was placed between the spot and condenser lenses, and an emission interference filter (Chroma D550/50 m) was placed as close as practical to the objective back focal plane.
In our case (as in Ref. [24]) the structured illumination image is obtained by projecting a speckle pattern in the sample, which is easily produced by introducing a diffuser plate (light shaping diffuser 10°, from kit 20DKIT-C1, Newport) in the laser beam path (laser Cobolt Calypso, ⌊ = 491 nm), in a plane conjugated with the microscope objective back-aperture.
We illuminated GECI-expressing pyramidal cells in hippocampal slice culture with 7 mW at the objective back aperture, and recorded the fluorescence intensity over a linescan image taken at the base of the apical dendrite (Figure 7; note that different filter sets were used for the FRET-based and the GFP-based GECIs; see Materials and Methods).
The power at the objective back aperture during ablation was 90 100 mW.
Laser powers were measured at the objective back aperture and corrected for objective transmission efficiency to represent the power at the sample.
