共查询到20条相似文献,搜索用时 15 毫秒
1.
Pircher M Kroisamer JS Felberer F Sattmann H Götzinger E Hitzenberger CK 《Biomedical optics express》2010,2(1):100-112
In this study we use our previously introduced scanning laser ophthalmoscope (SLO) / transverse scanning optical coherence tomography (TS-OCT) instrument to investigate long term changes in cone photoreceptors. The instrument is capable to provide 3D information of the human cone photoreceptors with negligible eye motion artifacts due to an implemented 3D motion correction on a cellular level. This allows for an in vivo investigation of exactly the same location on the retina with cellular resolution over several days. Temporal changes in the backscattered intensity are observed and quantified within the junction between inner and outer segments of cone photoreceptors, the cone outer segments, the end tips of cone photoreceptors and the retinal pigment epithelium. Furthermore, the length of individual cone outer segments is measured and observed over time. We show, to the best of our knowledge for the first time, that bright reflection spots which are located within the outer segment of cone photoreceptors change their position when observed over extended time periods. The average measured bright reflection spot motion speed corresponds well to the expected cone growth speed. We believe that this observation can be associated with the first direct in vivo imaging of the cone renewal process. 相似文献
2.
M. Pircher J. S. Kroisamer F. Felberer H. Sattmann E. G?tzinger C. K. Hitzenberger 《Biomedical optics express》2011,2(1):100-112
In this study we use our previously introduced scanning laser ophthalmoscope (SLO) / transverse scanning optical coherence tomography (TS-OCT) instrument to investigate long term changes in cone photoreceptors. The instrument is capable to provide 3D information of the human cone photoreceptors with negligible eye motion artifacts due to an implemented 3D motion correction on a cellular level. This allows for an in vivo investigation of exactly the same location on the retina with cellular resolution over several days. Temporal changes in the backscattered intensity are observed and quantified within the junction between inner and outer segments of cone photoreceptors, the cone outer segments, the end tips of cone photoreceptors and the retinal pigment epithelium. Furthermore, the length of individual cone outer segments is measured and observed over time. We show, to the best of our knowledge for the first time, that bright reflection spots which are located within the outer segment of cone photoreceptors change their position when observed over extended time periods. The average measured bright reflection spot motion speed corresponds well to the expected cone growth speed. We believe that this observation can be associated with the first direct in vivo imaging of the cone renewal process.OCIS codes: (170.4500) Optical coherence tomography, (330.5310) Vision – photoreceptors, (170.5755) Retina scanning, (170.4470) Ophthalmology, (170.2655) Functional monitoring and imaging 相似文献
3.
Franz Felberer Matthias Rechenmacher Richard Haindl Bernhard Baumann Christoph K. Hitzenberger Michael Pircher 《Biomedical optics express》2015,6(4):1407-1418
We use our previously developed adaptive optics (AO) scanning laser ophthalmoscope (SLO)/ optical coherence tomography (OCT) instrument to investigate its capability for imaging retinal vasculature. The system records SLO and OCT images simultaneously with a pixel to pixel correspondence which allows a direct comparison between those imaging modalities. Different field of views ranging from 0.8°x0.8° up to 4°x4° are supported by the instrument. In addition a dynamic focus scheme was developed for the AO-SLO/OCT system in order to maintain the high transverse resolution throughout imaging depth. The active axial eye tracking that is implemented in the OCT channel allows time resolved measurements of the retinal vasculature in the en-face imaging plane. Vessel walls and structures that we believe correspond to individual erythrocytes could be visualized with the system.OCIS codes: (170.3890) Medical optics instrumentation, (110.1080) Active or adaptive optics, (170.4470) Ophthalmology, (330.5310) Vision - photoreceptors, (110.4500) Optical coherence tomography 相似文献
4.
Robert J. Zawadzki Pengfei Zhang Azhar Zam Eric B. Miller Mayank Goswami Xinlei Wang Ravi S. Jonnal Sang-Hyuck Lee Dae Yu Kim John G. Flannery John S. Werner Marie E. Burns Edward N. Pugh Jr. 《Biomedical optics express》2015,6(6):2191-2210
Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a large-scale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.OCIS codes: (170.4460) Ophthalmic optics and devices, (110.4500) Optical coherence tomography, (110.1080) Active or adaptive optics, (170.0110) Imaging systems, (330.7324) Visual optics, comparative animal models, (170.4470) Ophthalmology 相似文献
5.
Kari V. Vienola Boy Braaf Christy K. Sheehy Qiang Yang Pavan Tiruveedhula David W. Arathorn Johannes F. de Boer Austin Roorda 《Biomedical optics express》2012,3(11):2950-2963
Fixational eye movements remain a major cause of artifacts in optical coherence tomography (OCT)
images despite the increases in acquisition speeds. One approach to eliminate the eye motion is to
stabilize the ophthalmic imaging system in real-time. This paper describes and quantifies the
performance of a tracking OCT system, which combines a phase-stabilized optical frequency domain
imaging (OFDI) system and an eye tracking scanning laser ophthalmoscope (TSLO). We show that active
eye tracking minimizes artifacts caused by eye drift and micro saccades. The remaining tracking lock
failures caused by blinks and large saccades generate a trigger signal which signals the OCT system
to rescan corrupted B-scans. Residual motion artifacts in the OCT B-scans are reduced to 0.32
minutes of arc (~1.6 µm) in an in vivo human eye enabling acquisition of
high quality images from the optic nerve head and lamina cribrosa pore structure.OCIS codes: (110.0110) Imaging systems, (110.4500) Optical coherence tomography, (170.4460) Ophthalmic optics and devices, (170.4470) Ophthalmology 相似文献
6.
Han Saem Cho Sun-Joo Jang Kyunghun Kim Alexey V. Dan-Chin-Yu Milen Shishkov Brett E. Bouma Wang-Yuhl Oh 《Biomedical optics express》2014,5(1):223-232
Intravascular optical frequency-domain imaging (OFDI), a second-generation optical coherence tomography (OCT) technology, enables imaging of the three-dimensional (3D) microstructure of the vessel wall following a short and nonocclusive clear liquid flush. Although 3D vascular visualization provides a greater appreciation of the vessel wall and intraluminal structures, a longitudinal imaging pitch that is several times bigger than the optical imaging resolution of the system has limited true high-resolution 3D imaging, mainly due to the slow scanning speed of previous imaging catheters. Here, we demonstrate high frame-rate intravascular OFDI in vivo, acquiring images at a rate of 350 frames per second. A custom-built, high-speed, and high-precision fiber-optic rotary junction provided uniform and high-speed beam scanning through a custom-made imaging catheter with an outer diameter of 0.87 mm. A 47-mm-long rabbit aorta was imaged in 3.7 seconds after a short contrast agent flush. The longitudinal imaging pitch was 34 μm, comparable to the transverse imaging resolution of the system. Three-dimensional volume-rendering showed greatly enhanced visualization of tissue microstructure and stent struts relative to what is provided by conventional intravascular imaging speeds.OCIS codes: (170.4500) Optical coherence tomography, (170.2150) Endoscopic imaging, (170.3880) Medical and biological imaging 相似文献
7.
Vimal Prabhu Pandiyan Xiaoyun Jiang James A. Kuchenbecker Ramkumar Sabesan 《Biomedical optics express》2021,12(9):5865
Line-scan OCT incorporated with adaptive optics (AO) offers high resolution, speed, and sensitivity for imaging retinal structure and function in vivo. Here, we introduce its implementation with reflective mirror-based afocal telescopes, optimized for imaging light-induced retinal activity (optoretinography) and weak retinal reflections at the cellular scale. A non-planar optical design was followed based on previous recommendations with key differences specific to a line-scan geometry. The three beam paths fundamental to an OCT system –illumination/sample, detection, and reference– were modeled in Zemax optical design software to yield theoretically diffraction-limited performance over a 2.2 deg. field-of-view and 1.5 D vergence range at the eye’s pupil. The performance for imaging retinal structure was exemplified by cellular-scale visualization of retinal ganglion cells, macrophages, foveal cones, and rods in human observers. The performance for functional imaging was exemplified by resolving the light-evoked optical changes in foveal cone photoreceptors where the spatial resolution was sufficient for cone spectral classification at an eccentricity 0.3 deg. from the foveal center. This enabled the first in vivo demonstration of reduced S-cone (short-wavelength cone) density in the human foveola, thus far observed only in ex vivo histological preparations. Together, the feasibility for high resolution imaging of retinal structure and function demonstrated here holds significant potential for basic science and translational applications. 相似文献
8.
Janet A. H. Tang Charles E. Granger Karteek Kunala Keith Parkins Khang T. Huynh Kristen Bowles-Johnson Qiang Yang Jennifer J. Hunter 《Biomedical optics express》2022,13(3):1737
The intrinsic fluorescence properties of lipofuscin – naturally occurring granules that accumulate in the retinal pigment epithelium – are a potential biomarker for the health of the eye. A new modality is described here which combines adaptive optics technology with fluorescence lifetime detection, allowing for the investigation of functional and compositional differences within the eye and between subjects. This new adaptive optics fluorescence lifetime imaging ophthalmoscope was demonstrated in 6 subjects. Repeated measurements between visits had a minimum intraclass correlation coefficient of 0.59 Although the light levels were well below maximum permissible exposures, the safety of the imaging paradigm was tested using clinical measures; no concerns were raised. This new technology allows for in vivo adaptive optics fluorescence lifetime imaging of the human RPE mosaic. 相似文献
9.
Sylvia Desissaire Florian Schwarzhans Stefan Steiner Clemens Vass Georg Fischer Michael Pircher Christoph K. Hitzenberger 《Biomedical optics express》2021,12(11):7092
We demonstrate the use of temporal phase evolution (TPE-) OCT methods to evaluate retinal tissue deformation in-vivo over time periods of several seconds. A custom built spectral domain (SD)-OCT system with an integrated retinal tracker, ensuring stable imaging with sub-speckle precision, was used for imaging. TPE-OCT measures and images phase differences between an initial reference B-scan and each of the subsequent B-scans of the evaluated temporal sequence. In order to demonstrate the precision and repeatability of the measurements, retinal nerve fiber (RNF) tissue deformations induced by retinal vessels pulsating with the heartbeat were analyzed in several healthy subjects. We show TPE maps (M-scans of phase evolution as a function of position along B-scan trace vs. time) of wrapped phase data and corresponding deformation maps in selected regions of the RNF layer (RNFL) over the course of several cardiac cycles. A reproducible phase pattern is seen at each heartbeat cycle for all imaged volunteers. RNF tissue deformations near arteries and veins up to ∼ 1.6 µm were obtained with an average precision for a single pixel of about 30 nm. Differences of motion induced by arteries and veins are also investigated. 相似文献
10.
Tu S Holm NR Koning G Huang Z Reiber JH 《The international journal of cardiovascular imaging》2011,27(2):197-207
The combination/fusion of quantitative coronary angiography (QCA) and intravascular ultrasound (IVUS)/optical coherence tomography
(OCT) depends to a great extend on the co-registration of X-ray angiography (XA) and IVUS/OCT. In this work a new and robust
three-dimensional (3D) segmentation and registration approach is presented and validated. The approach starts with standard
QCA of the vessel of interest in the two angiographic views (either biplane or two monoplane views). Next, the vessel of interest
is reconstructed in 3D and registered with the corresponding IVUS/OCT pullback series by a distance mapping algorithm. The
accuracy of the registration was retrospectively evaluated on 12 silicone phantoms with coronary stents implanted, and on
24 patients who underwent both coronary angiography and IVUS examinations of the left anterior descending artery. Stent borders
or sidebranches were used as markers for the validation. While the most proximal marker was set as the baseline position for
the distance mapping algorithm, the subsequent markers were used to evaluate the registration error. The correlation between
the registration error and the distance from the evaluated marker to the baseline position was analyzed. The XA-IVUS registration
error for the 12 phantoms was 0.03 ± 0.32 mm (P = 0.75). One OCT pullback series was excluded from the phantom study, since it did not cover the distal stent border. The
XA-OCT registration error for the remaining 11 phantoms was 0.05 ± 0.25 mm (P = 0.49). For the in vivo validation, two patients were excluded due to insufficient image quality for the analysis. In total
78 sidebranches were identified from the remaining 22 patients and the registration error was evaluated on 56 markers. The
registration error was 0.03 ± 0.45 mm (P = 0.67). The error was not correlated to the distance between the evaluated marker and the baseline position (P = 0.73). In conclusion, the new XA-IVUS/OCT co-registration approach is a straightforward and reliable solution to combine
X-ray angiography and IVUS/OCT imaging for the assessment of the extent of coronary artery disease. It provides the interventional
cardiologist with detailed information about vessel size and plaque size at every position along the vessel of interest, making
this a suitable tool during the actual intervention. 相似文献
11.
Betul Sahin Barbara Lamory Xavier Levecq Fabrice Harms Chris Dainty 《Biomedical optics express》2012,3(2):225-239
Adaptive optics, when integrated into retinal imaging systems, compensates for rapidly changing ocular aberrations in real time and results in improved high resolution images that reveal the photoreceptor mosaic. Imaging the retina at high resolution has numerous potential medical applications, and yet for the development of commercial products that can be used in the clinic, the complexity and high cost of the present research systems have to be addressed. We present a new method to control the deformable mirror in real time based on pupil tracking measurements which uses the default camera for the alignment of the eye in the retinal imaging system and requires no extra cost or hardware. We also present the first experiments done with a compact adaptive optics flood illumination fundus camera where it was possible to compensate for the higher order aberrations of a moving model eye and in vivo in real time based on pupil tracking measurements, without the real time contribution of a wavefront sensor. As an outcome of this research, we showed that pupil tracking can be effectively used as a low cost and practical adaptive optics tool for high resolution retinal imaging because eye movements constitute an important part of the ocular wavefront dynamics.OCIS codes: (110.1080) Active or adaptive optics, (100.4999) Pattern recognition, target tracking, (170.4460) Ophthalmic optics and devices, (170.3890) Medical optics instrumentation 相似文献
12.
Mengxi Zhou Nathan Doble Stacey S. Choi Tianyu Jin Chenwei Xu Srinivasan Parthasarathy Rajiv Ramnath 《Biomedical optics express》2022,13(10):5082
Adaptive optics imaging has enabled the enhanced in vivo retinal visualization of individual cone and rod photoreceptors. Effective analysis of such high-resolution, feature rich images requires automated, robust algorithms. This paper describes RC-UPerNet, a novel deep learning algorithm, for identifying both types of photoreceptors, and was evaluated on images from central and peripheral retina extending out to 30° from the fovea in the nasal and temporal directions. Precision, recall and Dice scores were 0.928, 0.917 and 0.922 respectively for cones, and 0.876, 0.867 and 0.870 for rods. Scores agree well with human graders and are better than previously reported AI-based approaches. 相似文献
13.
Geng Y Dubra A Yin L Merigan WH Sharma R Libby RT Williams DR 《Biomedical optics express》2012,3(4):715-734
Correction of the eye's monochromatic aberrations using adaptive optics (AO) can improve the resolution of in vivo mouse retinal images [Biss et al., Opt. Lett. 32(6), 659 (2007) and Alt et al., Proc. SPIE 7550, 755019 (2010)], but previous attempts have been limited by poor spot quality in the Shack-Hartmann wavefront sensor (SHWS). Recent advances in mouse eye wavefront sensing using an adjustable focus beacon with an annular beam profile have improved the wavefront sensor spot quality [Geng et al., Biomed. Opt. Express 2(4), 717 (2011)], and we have incorporated them into a fluorescence adaptive optics scanning laser ophthalmoscope (AOSLO). The performance of the instrument was tested on the living mouse eye, and images of multiple retinal structures, including the photoreceptor mosaic, nerve fiber bundles, fine capillaries and fluorescently labeled ganglion cells were obtained. The in vivo transverse and axial resolutions of the fluorescence channel of the AOSLO were estimated from the full width half maximum (FWHM) of the line and point spread functions (LSF and PSF), and were found to be better than 0.79 μm ± 0.03 μm (STD)(45% wider than the diffraction limit) and 10.8 μm ± 0.7 μm (STD)(two times the diffraction limit), respectively. The axial positional accuracy was estimated to be 0.36 μm. This resolution and positional accuracy has allowed us to classify many ganglion cell types, such as bistratified ganglion cells, in vivo. 相似文献
14.
15.
Souvick Mukherjee Tharindu De Silva Peyton Grisso Henry Wiley D. L. Keenan Tiarnan Alisa T Thavikulwat Emily Chew Catherine Cukras 《Biomedical optics express》2022,13(6):3195
Introduction – Retinal layer segmentation in optical coherence tomography (OCT) images is an important approach for detecting and prognosing disease. Automating segmentation using robust machine learning techniques lead to computationally efficient solutions and significantly reduces the cost of labor-intensive labeling, which is traditionally performed by trained graders at a reading center, sometimes aided by semi-automated algorithms. Although several algorithms have been proposed since the revival of deep learning, eyes with severe pathological conditions continue to challenge fully automated segmentation approaches. There remains an opportunity to leverage the underlying spatial correlations between the retinal surfaces in the segmentation approach. Methods - Some of these proposed traditional methods can be expanded to utilize the three-dimensional spatial context governing the retinal image volumes by replacing the use of 2D filters with 3D filters. Towards this purpose, we propose a spatial-context, continuity and anatomical relationship preserving semantic segmentation algorithm, which utilizes the 3D spatial context from the image volumes with the use of 3D filters. We propose a 3D deep neural network capable of learning the surface positions of the layers in the retinal volumes. Results - We utilize a dataset of OCT images from patients with Age-related Macular Degeneration (AMD) to assess performance of our model and provide both qualitative (including segmentation maps and thickness maps) and quantitative (including error metric comparisons and volumetric comparisons) results, which demonstrate that our proposed method performs favorably even for eyes with pathological changes caused by severe retinal diseases. The Mean Absolute Error (MAE) and Root Mean Squared Error (RMSE) for patients with a wide range of AMD severity scores (0–11) were within 0.84±0.41 and 1.33±0.73 pixels, respectively, which are significantly better than some of the other state-of-the-art algorithms. Conclusion – The results demonstrate the utility of extracting features from the entire OCT volume by treating the volume as a correlated entity and show the benefit of utilizing 3D autoencoder based regression networks for smoothing the approximated retinal layers by inducing shape based regularization constraints. 相似文献
16.
The present study aimed to develop and validate an advanced image stitching algorithm integrated with chemical imaging at the nanometre scale. This was applied to track the swelling, erosion, drug release and changes in surface texture of a swelling-controlled release system. The technique involves the delivery and withdrawal of a liquid droplet from the surface of the tablet alongside capturing multiple images of tablet surface using white light profilometry. The recovered liquid was then subject to chemical analysis for the quantification of drug and HPMC. The multiple images acquired during drug release were stitched together using an algorithm developed to generate a full tablet surface. New methods for swelling analysis (regional point, area and multiple regional analysis techniques) were also successfully developed. The results exhibited the exceptional capability of this technique for providing quantitative information regarding swelling, erosion, drug release and surface topography, hence negating the need for separate investigations. Moreover, it can also be anticipated that this technique may have potential use in other fields where surface dissolution, erosion and swelling have significant impact.Nanoscale chemical imaging technique for simultaneous quantification of swelling, erosion, drug release and 3D surface topography with full surface scanning. 相似文献
17.
Yoo YM Sikdar S Karadayi K Kolokythas O Kim Y 《Ultrasound in medicine & biology》2008,34(8):1221-1231
In three-dimensional (3D) ultrasound color Doppler imaging (CDI), effective rejection of flash artifacts caused by tissue motion (clutter) is important for improving sensitivity in visualizing blood flow in vessels. Since clutter characteristics can vary significantly during volume acquisition, a clutter rejection technique that can adapt to the underlying clutter conditions is desirable for 3D CDI. We have previously developed an adaptive clutter rejection (ACR) method, in which an optimum filter is dynamically selected from a set of predesigned clutter filters based on the measured clutter characteristics. In this article, we evaluated the ACR method with 3D in vivo data acquired from 37 kidney transplant patients clinically indicated for a duplex ultrasound examination. We compared ACR against a conventional clutter rejection method, down-mixing (DM), using a commonly-used flow signal-to-clutter ratio (SCR) and a new metric called fractional residual clutter area (FRCA). The ACR method was more effective in removing the flash artifacts while providing higher sensitivity in detecting blood flow in the arcuate arteries and veins in the parenchyma of transplanted kidneys. ACR provided 3.4 dB improvement in SCR over the DM method (11.4 +/- 1.6 dB versus 8.0 +/- 2.0 dB, p < 0.001) and had lower average FRCA values compared with the DM method (0.006 +/- 0.003 versus 0.036 +/- 0.022, p < 0.001) for all study subjects. These results indicate that the new ACR method is useful for removing nonstationary tissue motion while improving the image quality for visualizing 3D vascular structure in 3D CDI. 相似文献
18.
Masahiro Yamanari Satoko Nagase Shinichi Fukuda Kotaro Ishii Ryosuke Tanaka Takeshi Yasui Tetsuro Oshika Masahiro Miura Yoshiaki Yasuno 《Biomedical optics express》2014,5(5):1391-1402
The relationship between scleral birefringence and biometric parameters of human eyes in vivo is investigated. Scleral birefringence near the limbus of 21 healthy human eyes was measured using polarization-sensitive optical coherence tomography. Spherical equivalent refractive error, axial eye length, and intraocular pressure (IOP) were measured in all subjects. IOP and scleral birefringence of human eyes in vivo was found to have statistically significant correlations (r = −0.63, P = 0.002). The slope of linear regression was −2.4 × 10−2 deg/μm/mmHg. Neither spherical equivalent refractive error nor axial eye length had significant correlations with scleral birefringence. To evaluate the direct influence of IOP to scleral birefringence, scleral birefringence of 16 ex vivo porcine eyes was measured under controlled IOP of 5−60 mmHg. In these ex vivo porcine eyes, the mean linear regression slope between controlled IOP and scleral birefringence was −9.9 × 10−4 deg/μm/mmHg. In addition, porcine scleral collagen fibers were observed with second-harmonic-generation (SHG) microscopy. SHG images of porcine sclera, measured on the external surface at the superior side to the cornea, showed highly aligned collagen fibers parallel to the limbus. In conclusion, scleral birefringence of healthy human eyes was correlated with IOP, indicating that the ultrastructure of scleral collagen was correlated with IOP. It remains to show whether scleral collagen ultrastructure of human eyes is affected by IOP as a long-term effect.OCIS codes: (170.4500) Optical coherence tomography, (170.4470) Ophthalmology, (050.2555) Form birefringence, (180.4315) Nonlinear microscopy 相似文献
19.
Intrinsic optical signal (IOS) imaging promises a noninvasive method for objective assessment of retinal function. This study demonstrates concurrent optical coherence tomography (OCT) of amplitude-IOS and phase-IOS changes in human photoreceptors. A new procedure for differential-phase-mapping (DPM) is validated to enable depth-resolved phase-IOS imaging. Dynamic OCT revealed rapid amplitude-IOS and phase-IOS changes, which occur almost right away after the stimulus onset. These IOS changes were predominantly observed within the photoreceptor outer segment (OS), particularly two boundaries connecting to the inner segment and retinal pigment epithelium. The comparative analysis supports that both amplitude-IOS and phase-IOS attribute to transient OS morphological change associated with phototransduction activation in retinal photoreceptors. A simulation modeling is proposed to discuss the relationship between the photoreceptor OS length and phase-IOS changes. 相似文献
20.
The performance of structured illumination microscopy (SIM) systems depends on the computational method used to process the raw data. In this paper, we present a regularized three-dimensional (3D) model-based (MB) restoration method with positivity constraint (PC) for 3D processing of data from 3D-SIM (or 3-beam interference SIM), in which the structured illumination pattern varies laterally and axially. The proposed 3D-MBPC method introduces positivity in the solution through the reconstruction of an auxiliary function using a conjugate-gradient method that minimizes the mean squared error between the data and the 3D imaging model. The 3D-MBPC method provides axial super resolution, which is not the same as improved optical sectioning demonstrated with model-based approaches based on the 2D-SIM (or 2-beam interference SIM) imaging model, for either 2D or 3D processing of a single plane from a 3D-SIM dataset. Results obtained with our 3D-MBPC method show improved 3D resolution over what is achieved by the standard generalized Wiener filter method, the first known method that performs 3D processing of 3D-SIM data. Noisy simulation results quantify the achieved 3D resolution, which is shown to match theoretical predictions. Experimental verification of the 3D-MBPC method with biological data demonstrates successful application to data volumes of different sizes. 相似文献