Figure 3
From: Three-Dimensional Microscopy by Milling with Ultraviolet Excitation
Monte-Carlo simulations of confocal and MUVE point-spread-functions using coupled-wave theory for absorbance in a layered homogeneous substrate22. All simulations show x-polarized coherent light propagating from left to right and intensities are normalized for each image. Contours indicate (from darkest to lightest) 1%, 10%, and 30% thresholds of maximum intensity. (red) Confocal PSFs for imaging in idealized (i.e. cleared) samples using 0.4NA (left), 0.8NA (center), and 1.0NA (right) objectives. In MUVE imaging, exponential absorbance of the excitation is the dominant factor describing the axial PSF. (purple) Incident deep-UV light is shown incident on a sample using a low-NA (≈0.25) objective. Varying the molar absorbance by doping the embedding compound reduces penetration, creating a smaller axial PSF. A UV-transparent sample (left) shows a significant contribution from back-scattered light. However, doping with UV27 (center, right) results in a significant improvement over high-end confocal imaging. (green) The excited region of the samples scales with the penetrating UV PSF, however ablation results in truncation of the left half. (bottom) A comparison is shown between confocal and MUVE axial PSFs. Lateral resolution is theoretically identical between MUVE and confocal.
