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Results: 5

1.
Fig. 4

Fig. 4. From: Adaptive optics enables 3D STED microscopy in aberrating specimens.

Images of 100 nm fluorescent beads through ~55 µm layer of glycerol. xz images were acquired in confocal mode (A) before and (B) after correction of sample induced aberrations, and STED mode with (C) correction to the excitation beam only and (D) correction to excitation and depletion beam paths. (E) Axial line profiles of pixels summed across horizontal dimension of dashed boxes in (C) in (D) show improvement in both resolution and signal when aberrations are corrected in the STED beam path.

Travis J. Gould, et al. Opt Express. 2012 September 10;20(19):20998-21009.
2.
Fig. 3

Fig. 3. From: Adaptive optics enables 3D STED microscopy in aberrating specimens.

Adaptive correction of residual system aberrations. (A, B) STED PSF using the central λ/2 phase mask before (A) and after (B) correction for system aberrations with corresponding phase patterns used on SLM2. (C, D) xz STED images of 100 nm fluorescent bead attached to coverglass and imaged (C) before and (D) after correction of system aberrations. (E) Axial line profiles of pixels summed across horizontal dimension of dashed boxes in (C) and (D). Fitted curves give FWHM of 280 nm (Gaussian fit) for the uncorrected STED image and 140 nm (Lorentzian fit) when adaptive optics is used to compensate for system-induced aberrations in the STED beam path.

Travis J. Gould, et al. Opt Express. 2012 September 10;20(19):20998-21009.
3.
Fig. 2

Fig. 2. From: Adaptive optics enables 3D STED microscopy in aberrating specimens.

Metric curves as a function of the SLM-applied aberration for correction in the depletion beam path when using a central λ/2 phase mask. Example curves are shown for Zernike modes 5 (astigmatism), 7 (coma), 9 (trefoil), 11 (1st spherical), 22 (2nd spherical), and 37 (3rd spherical) including sharpness (S; green lines), brightness (B; black lines), and combined (M; red lines) metrics. A prime (e.g. Z11) indicates that a particular mode was corrected to be displacement free. Insets show corresponding normalized (-π to + π) phase distributions in the objective back aperture. Data was obtained by imaging 200 nm fluorescent beads in STED mode.

Travis J. Gould, et al. Opt Express. 2012 September 10;20(19):20998-21009.
4.
Fig. 5

Fig. 5. From: Adaptive optics enables 3D STED microscopy in aberrating specimens.

AO STED images of fluorescent beads through zebrafish retina sections. (A-F) Results for beads imaged though ~14 µm of retina. Lateral and axial sections of a single fluorescent bead imaged in (A, D) confocal, (B, E) STED, and (C, F) AO STED show improvement in signal and resolution when adaptive aberration correction is applied to the depletion beam path. (G-L) Similar image sequences for beads imaged through ~25 µm of retina. Axial profiles of beads in AO STED images were ~208 nm and ~249 nm for (F) and (L), respectively. Color bar in (E) also applies to (B), (C), and (F). Color bar in (K) also applies to (H), (I), and (L). (M-O) Volume renderings for data shown in (A-F) for (M) confocal, (N) STED, and (O) AO STED data. (N) and (O) plotted on same color scale for comparison of signal.

Travis J. Gould, et al. Opt Express. 2012 September 10;20(19):20998-21009.
5.
Fig. 1

Fig. 1. From: Adaptive optics enables 3D STED microscopy in aberrating specimens.

Schematic of AO STED setup. FI - Faraday isolator; GLP - Glan laser polarizer; GB - glass block; DS - delay stage; AOM - acousto-optical modulator; PMF - polarization-maintaining fiber; SLM1/2 - spatial light modulators; PC - photonic crystal fiber; AOTF - acousto-optical tunable filter; PBS - polarizing beam splitter cube; λ/2 - half-wave plate; λ/4 - quarter-wave plate; DM1/2 - dichroic mirrors; F - bandpass filter; MMF - multimode fiber; APD - avalanche photodiode; OBJ - objective lens; xyz - 3 axes piezo sample stage. Insets show typical phase patterns for each SLM where phase contributions include a baseline flatness correction for the SLM (provided by the manufacturer), a circular blazed grating (for off-axis phase modulation) that defines the active area, correction for system-induced aberration, and a central λ/2 phase mask (SLM1 only).

Travis J. Gould, et al. Opt Express. 2012 September 10;20(19):20998-21009.

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