On the basis of the immunostaining results, this work provided convincing evidence for the concurrent appearance of NET components of citrullinated histone, MPO and PAD4 in necrotizing lesions of the early focal phase of ANCA-associated polyangiitis.
Immunostaining of renal biopsy revealed that NET components of citrullinated histone, MPO and PAD4 were concurrently deposited both around fibrinoid necrosis in necrotizing glomerulonephritis in the early focal phase of ANCA-associated polyangiitis and along an interlobular arterial wall.
Remarkably similar elongated tubules were also seen by the focal fluid-phase endocytosis assay, validating the endocytic nature of these compartments.
To further monitor the subsequent transport and fate of nascent endocytic compartments, we performed the focal fluid-phase endocytic assay while extending the duration of time-lapse confocal imaging.
Then a weak modulation of the point spread function (PSF) is generated at the focal plane, allowing for direct phase measurements.
The image of the illumination function depicts a cross section of the amplitude and phase within the focal plane of the beam and, thus, determines the final spot profile at the sample surface.
Following convention, we express the variation of phase near the focal plane for small values of defocus as θ ≈ (4 πn/ λ) f−1Δ z where f−1 is a constant factor [ 47].
The performances of the designed OFPEs are appraised in detail, for instance, the line-width of the transmissive peaks, the spot size of the focusing point on the focal plane, and the effect of phase quantized level of the OFPEs, etc.
Since the DMD is placed on the image-conjugate plane of the objective lens' focal plane, a sinusoidal MPE pattern with different rotations and phase shifts can be precisely projected on the focal plane.
The polymeric phase plate (PP) induced a clockwise 2π-phase shift across the STED beam, which generated the doughnut shaped intensity distribution in the focal plane.
