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Here a switchable acoustic resolution (AR) and optical resolution (OR) photoacoustic microscopy (AR-OR-PAM) system capable of both high resolution imaging at shallow depth and low resolution deep tissue imaging on the same sample in vivo is demonstrated.
Photoacoustic microscopy (PAM) is a fast-growing invivo imaging modality that combines both optics and ultrasound, providing penetration beyond the optical mean free path (~1 mm in skin) with high resolution. By combining optical absorption contrast with the high spatial resolution of ultrasound in a single modality, this technique can penetrate deep tissues. Photoacoustic microscopy systems can have either a low acoustic resolution and probe deeply or a high optical resolution and probe shallowly. It is challenging to achieve high spatial resolution and large depth penetration with a single system. This work presents an AR-OR-PAM system capable of both high-resolution imaging at shallow depths and low-resolution deep-tissue imaging of the same sample in vivo. A lateral resolution of 4 µm with 1.4 mm imaging depth using optical focusing and a lateral resolution of 45 µm with 7.8 mm imaging depth using acoustic focusing were successfully demonstrated using the combined system. Here, in vivo small-animal blood vasculature imaging is performed to demonstrate its biological imaging capability.
High-resolution optical imaging modalities, such as optical coherence tomography, confocal microscopy, and multiphoton microscopy, have numerous benefits. However, the spatial resolution decreases significantly as the imaging depth increases. This is because of the diffuse nature of light transport in soft tissues1,2. The integration of optical excitation and ultrasound detection provides a solution to overcome the challenge of high-resolution optical imaging in deep tissues. Photoacoustic microscopy (PAM) is one such modality that can provide deeper imaging than other optical imaging modalities. It has been s....
All animal experiments were performed according to the approved regulations and guidelines of the Institutional Animal Care and Use Committee of Nanyang Technological University, Singapore (Animal Protocol Number ARF-SBS/NIE-A0263).
1. AR-OR-PAM System (Figure 1)
The schematic of the AR-OR-PAM system is shown in Figure 1. In this setup, all components were integrated and assembled in an optical cage setup. The use of a cage system makes the AR-OR-PAM scanning head compact and easily assembled, aligned, and integrated onto a single scanning stage.
Two-dimensional continuous raster scanning of the imaging head was used during image acquisition. The time-resolv.......
In conclusion, a switchable AR and OR PAM system that can achieve both high-resolution imaging at lower imaging depths and lower-resolution imaging at higher imaging depths has been developed. The lateral resolution and imaging depth of the switchable system was determined. The advantages of this switchable PAM system include: (1) the high-resolution imaging using tight optical focusing; (2) the deep-tissue imaging using acoustic focusing; 3) the dark-field illumination for AR-PAM, which prevents strong PA signals from a.......
All animal experiments were performed according to the approved guidelines and regulations of the Institutional Animal Care and Use Committee of Nanyang Technological University, Singapore (Animal Protocol Number ARF-SBS/NIE-A0263). The authors have no relevant financial interests in the manuscript and no other potential conflicts of interest to disclose.
The authors would like to acknowledge the financial support from a Tier 2 grant funded by the Ministry of Education in Singapore (ARC2/15: M4020238). The authors would also like to thank Mr. Chow Wai Hoong Bobby for the machine shop help.
....Name | Company | Catalog Number | Comments |
Q-switched Nd:YAG laser | Edgewave | BX80-2-L | Pump laser |
Credo-High Repetition Rate Dye Laser | Spectra physics | CREDO-DYE-N | Dye laser |
Precision Linear Stage | Physik Instrumente | PLS 85 | XY raster scanning stage |
Translation stage | Physik Instrumente | VT 80 | Confocal determine |
Mounted Silicon photodiode | Thorlabs | SM05PD1A | Triggering/Pulse variation |
Motorized continuous Rotational stage | Thorlabs | CR1/M-Z7 | Diverting laser beam |
Mounted Continuously Variable ND Filter | Thorlabs | NDC-50C-4M | Intensity variable |
Fiber Patch Cable | Thorlabs | M29L01 | Multimode fiber |
Microscope objective | Newport | M-10X | Objective |
XY translating mount | Thorlabs | CXY1 | Translating mount |
Plano convex lens | Thorlabs | LA1951 | Collimating lens |
Conical lens | Altechna | APX-2-B254 | Ring shape beam |
Translation stage | Thorlabs | CT1 | Translating stage |
Optical condenser | Home made | ||
Ultrasonic transducer | Olympus-NDT | V214-BB-RM | 50MHz transducer |
Plano concave lens | Thorlabs | LC4573 | Acoustic lens |
Pulser/Receiver | Olympus-NDT | 5073PR | Pulse echo amplifier |
Mounted standard iris | Thorlabs | ID12/M | Beam shaping |
Plano convex lens | Thorlabs | LA4327 | Condenser lens |
Mounted precision pinhole | Thorlabs | P50S | Spatial filtering |
Single mode fiber patch cable | Thorlabs | P1-460B-FC-1 | Single mode fiber |
Fiber coupler | Newport | F-91-C1 | Single mode coupling |
Achromatic doublet lens | Edmund Optics | 32-317 | Achromatic doublet |
Protected silver elliptical mirror | Thorlabs | PFE10-P01 | Mirror |
Right angle kinematic mirror mount | Thorlabs | KCB1 | Mirror mount |
Z-Axis Translation Mount | Thorlabs | SM1Z | z translator |
Lens tube | Thorlabs | SM05L10 | |
UV Fused Silica Right-Angle Prism | Thorlabs | PS615 | Right angle prism |
Rhomboid prism | Edmund Optics | 47-214 | Shear wave |
Dimethylpolysiloxane | Sigma Aldrich | DMPS1M | Silicon oil |
Amplifier | Mini Circuits | ZFL-500LN | Amplifier |
16 bit high speed digitizer | Spectrum | M4i.4420 | Data acquisition card |
Oscilloscope | Agilent Technologies | DS06014A | |
Mice | InVivos Pte.Ltd | ICR | Animal model |
Ultrasound gel | Progress/parker acquasonic gel | PA-GEL-CLEA-5000 | Acoustic coupling |
Water tank | Home made | ||
Translation stage | Homemade | Switching AR-OR | |
Gold nanoparticles | Sigma Aldrich | 742031 | Lateral resolution |
Sterile ocular ointment | Alcon | Duratears | Animal imaging |
1951 USAF resolution test target | Edmund Optics | 38257 | Confocal alignment |
Data acquisition software | National Instrument | Labview | Home made software using Labview |
Image Processing software | Mathworks | Matlab | Home made program using Matlab |
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