Session - Biosensing and Bio-Manipulation Techniques II (BW2A): paper BW2A.7Bio-Optics: Design and Application (BODA)We present experimental demonstration of pixel super-resolution time-stretch imaging by high-speed agile-beam-steering with the use of synchronized acousto-optic deflector--enabling high-resolution imaging rate of 1MHz whereas relaxing the stringent requirement on extreme data acquisition. © 2015 OSApostprin

Chan, ACS

Lam, EY

Tsia, KK

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Title Pixel super-resolution in optical time-stretch microscopy usingacousto-optic deflectorAuthor(s) Chan, ACS; Lam, EY; Tsia, KKCitationThe 2015 Optics in the Life Sciences Congress, Vancouver,Canada, 12-15 April 2015. In Conference Proceedings, 2015,paper BW2A.7Issued Date 2015URL http://hdl.handle.net/10722/211413Rights© 2015 Optical Society of America. One print or electronic copymay be made for personal use only. Systematic reproductionand distribution, duplication of any material in this paper for afee or for commercial purposes, or modifications of the contentof this paper are prohibited.BW2A.7.pdf Optics in the Life Sciences 2015 © OSA 2015Pixel super-resolution in optical time-stretchmicroscopy using acousto-optic deflectorAntony C. S. Chan, Edmund Y. Lam and Kevin K. Tsia∗Department of Electrical and Electronic Engineering, The University of Hong Kong, Pokfulam, Hong Kong∗tsia@hku.hkAbstract: We present experimental demonstration of pixel super-resolution time-stretchimaging by high-speed agile-beam-steering with the use of synchronized acousto-opticdeflector – enabling high-resolution imaging rate of 1MHz whereas relaxing the stringentrequirement on extreme data acquisition.© 2015 Optical Society of AmericaOCIS codes: 100.0118, 100.6640, 170.3880Enabling ultra-high continuous frame/line-scan rate (as high as multi-MHz and even more), optical time-stretchimaging is proven to be an unique potential solution for high-throughput image-based bioassay with high-contentsingle-cell image information (e.g. morphological information, quantitative phase and and absorption contrast [1–3]).Operating at an ultrafast imaging rate, however, entails the need for high-speed, high-bandwidth digitizer (> 40 GSa/ssampling rate, > 10 GHz analog bandwidth) to guarantee sufficient time-stretch image pixel resolution. Such state-of-the-art specification might prohibit the wide-spread utilization of this imaging technology for practical biomedicaldiagnostics. To address this challenge, we have previously proposed to adopt the concept of pixel super-resolution(SR) to time-stretch imaging with the aim of relaxing the stringent requirement on high-bandwidth digitizer whereasthe high image resolution can still be preserved [4]. Common in conventional photography/imaging, pixel SR imagingis employed to circumvent the pixel-size limitation (or equivalently to improve spatial-sampling bandwidth) by fusingmultiple subpixel-shifted, low-sampling-rate (LR) frames with minimal reduction in the effective frame rate [6]. Thekey technical challenge of adopting pixel SR technique with time-stretch imaging is to implement ultrafast sub-pixelshifts in order to maintain the high frame rate on the order of 1 MHz. In this paper, we experimentally demonstratethat such ultrahigh-speed sub-pixel shift (in the multi-MHz regime) can be achieved by synchronized high-precisionbeam steering through the use of acousto-optic deflector (AOD) – realizing practical pixel SR time-stretch imagingsystem at a single-shot line-scan rate of 1 MHz.Fig. 1 shows the schematics of an AOD-enabled pixel-SR time-stretch imaging. A broadband pulsed laser (centeredat 1064nm) is time-stretched by a dispersive fiber before feeding into the microscopic setup in free space. The pulsedDigital frequencysynthesizer (DDSPA) Custom-madecontroller boardAODAchromatic doublet pairObjective lens DiffractiongratingLaser beamBeam stop BeamsplitterReflected1st order beamResolutiontargetTime-stretched pulsedbroadband laser To photodetector& digitizer Fiber collimator(b)(a)0th order1st order(c)frequencychirpAODFig. 1: Schematics of AOD-enabled pixel super-resolution (SR) time-stretch imaging. Subpixel beam steering is re-alized by feeding an microwave frequency chirp to the AOD. For the sake of clarity, only the laser beam at centerwavelength (1060nm) is shown here. (a) Subpixel-shifted laser beam of single wavelength component; (b) Zoom-inview of spectral-encoding process with multiple wavelengths; (c) exaggerated view of Bragg diffraction by the AOD.BW2A.7.pdf Optics in the Life Sciences 2015 © OSA 2015global time / ns0 100 200 300 400 500 600 700 800ADCvalue/A.U.Digital frequency control signal for AOD}STEAM line scan signal109.50MHz 109.98MHz 110.47MHz 110.96MHz(b)(c)(a)(d)Fig. 2: Four low-sampling-rate (LR) frames are captured by precisely controlling the subpixel shift by the AOD. (a)Single-shot time-stretch image in synchronous with the beam steering signals driving the AOD (i.e. the yellow andorange traces representing the binary frequency control signals (00,01,10,11)). The 8 consecutive pulses are pulse-averaged to produce (b) 4 subpixel-shifted LR frames, which are then interleaved to produce (c) a pixel SR time-stretchimage; (d) A bright-field image of the USAF target group 8 and 9 is captured for comparison.beam passes through the AOD which performs agile beam-steering at the diffraction angle directly proportional to thedriving acoustic frequency onto the AOD (Fig. 1a). This beam steering is solely responsible for the ultrafast sub-pixelshifts, which will be discussed shortly. The diffraction grating, positioned at the back aperture plane of the objectivelens via the achromatic doublet pair, disperses different wavelengths of the laser beam to the corresponding lateralposition on the object plane – a process called spectral-encoding (Fig. 1b). The reflected beam is descanned by thesame diffraction grating along the same path, and is coupled into a single-mode fiber which serves as a confocal pinholeof time-stretch imaging system. Then, time-stretch-encoded signal is relayed to the photodetector and the high-speeddigitizer for image acquisition. To reject the unwanted zeroth order beam from the AOD, a beam stop is placed atthe image plane between the achromatic doublet pair. The field-of-view of the STEAM system is limited to 90µmgovened by the total bandwidth of the laser source (12 nm). To match optical resolution limit at around 2.2µm, a highspeed digitizer with an analog bandwidth > 16 GHz, sampling rate 80 GSa/s is required. Here, we demonstrate thatthe same performance can be achieved from a digitizer with lower bandwidth (8 GHz) and sampling rate (20 GSa/s)by synchronous subpixel-shift control of the AOD.To synchronize the AOD to the home-built pulsed fiber laser, a custom-designed printed circuit board is fabricated tophase-lock the pulse output. This is to produce a set of binary frequency control signals (00, 01, 10, 11) for an digitalfrequency synthesizer (AA Opto-electronic DDSPA-B15b), as shown in Fig. 2a. As a result, every 2 consecutivepules encode one set of subpixel-shifted low-sampling-rate (LR) line-scan. In total, 8 consecutive pulses complete onesubpixel-shift imaging cycle. To meet these requirements, the beam has to settle at a subpixel shift of 2.2µm/4 =0.5µm within the pulse repetition interval of 89 ns. This is feasible with an acoustic frequency chirp ∆f = 0.49 MHzwith a beam settle time less than 40 ns [7], as shown in Fig. 2. This results in an effective line-scanning frame rate ashigh as 1 MHz. To reconstruct a pixel SR image from multiple LR frames, the 8 consecutive spectrally-encoded pulsesare pulse-averaged to 4 LR frames (Fig. 2b), and then fused by interleaving the pixelated samples (Fig. 2c). The twodimensional LR image is captured by scanning the sample stage orthogonal to the beam steering axis with the use of amotorized actuator (Newport LTA-HL) We note that this orthogonal scanning can be replaced by active beam steeringcontrol by another AOD, or other high-speed motions such as ultrafast microfludic flow [3].Fig. 3 shows the zoom-in view of the pixel SR time-stretch image, in comparison with the same region captured bythe selected LR frame. It is clear that pixel SR is able to recover the feature of Elements 5 and 6 (E5 and E6), whichcorrespond to a spatial resolution down to 2.2µm. Judging from the line profile of the high-sampling-rate (HR) image,we verify that the image resolution is not limited by the sampling rate (80 GSa/s) and analog bandwidth (16 GHz) ofthe digitizer. In contrast, when the digitizer is down-graded (20 GSa/s sample rate, 8 GHz analog bandwidth), the LRframe appears to be highly pixelated and the fine image feature is lost. Such loss is caused by the unfavorable subpixelposition at which the LR frame is registered, that fails to record the fine features. Pixel SR manages to restore thelost signal bandwidth by multiple subpixel-shift acquisitions, thus guarantees that at least one LR frame can partiallyregister the fine features. Unlike the conventional imaging concept, the significance here is that the sub-pixel time-stretch image shifts is performed at an ultrafast rate (i.e. in the MHz regime) and with the timing precision as shortas sub-nanoseconds (see Fig. 2). This is made possible by the presented high-speed synchronized agile-beam steeringwith the use of AOD. The image contrast of Element 5 (∼ 28 ADC steps) is just below the contrast of correspondingBW2A.7.pdf Optics in the Life Sciences 2015 © OSA 2015(a)(b)local time / nsx-axis / umE6 E5 E4 E3 E2(c)(d)0 1 2 3 48-bit ADC valuePixel SRLRHR, 80GSa/s(a)(b)(c)HR, 80GSa/sSR, 4 X 20GSa/sLR, 20GSa/s0 10 20 30 40 50 60 70 80 90-150-100-50050100150200250Fig. 3: Close up view of USAF target Group 8, showing an improved contrast of pixel SR time-stretch image over LRimage. (a) An LR frame acquired at 20 GSa/s sampling rate, 8 GHz analog bandwidth. (b) Pixel SR image generatedfrom four subpixel-shifted LR frames; (c) High-sampling-rate (HR) image acquired at 80 GSa/s, 16 GHz analogbandwidth; (d) line profile showing an improvement of contrast, particularly in Elements 5 and 6 (E5 and E6).feature (∼ 39 ADC steps) in a high-sample-rate (HR) image (Fig. 3d) due to a lower analog bandwidth. Nevertheless,this seemingly-lost bandwidth can be recovered by image sharpening technique, present in all advanced pixel SRalgorithm [6].In summary, we have demonstrated pixel SR time-stretch imaging enabled by agile-beam-steering with the use of asynchronized AOD, at a high-speed sub-pixel shift cycle in the MHz regime and with a sub-nanosecond timing preci-sion. This proof-of-principle demonstration (effective pixel SR line-scan rate of 1MHz) shows promising applicationof time-stretch imaging to high-throughput and high-content image-based cellular bioassay, particularly in the formatof microfluidic flow, without resorting to the state-of-the-art high-speed back-end data acquisition.This work is partially supported by grants from the Research Grants Council of Hong Kong Special AdministrativeRegion, China (HKU 717911E, HKU 720112E, 17207714 and 102009399), ITS/090/14 Innovation and TechnologySupport Programme (Tier 3) and the University Development Fund of HKU.References1. K. Goda, K. K. Tsia, and B. Jalali, Nature 458, 1145–9 (2009).2. A. K. S. Lau, T. T. W. Wong, A. C. S. Chan, E. Y. Lam, K. K. Y. Wong, and K. K. Tsia, Optics in the LifeSciences 1, NW1B.4 (2013).3. T. T. W. Wong, A. K. S. Lau, K. K. Y. Ho, M. Y. H. Tang, J. D. F. Robles, X. Wei, A. C. S. Chan, A. H. L. Tang,E. Y. Lam, K. K. Y. Wong, G. C. F. Chan, H. C. Shum, and K. K. Tsia, Scientific reports 4, 3656 (2014).4. T. T. W. Wong, Y. Qiu, A. K. S. Lau, J. Xu, A. C. S. Chan, K. K. Y. Wong, and K. K. Tsia, 8553, 85,531P(2012).5. T. T. W. Wong, A. C. S. Chan, K. K. Y. Wong, and K. K. Tsia, Conference on Lasers and Electro-Optics 20121, 1–2 (2012).6. S. Park, M. Park, and M. Kang, Signal Processing Magazine pp. 21–36 (2003).7. A. A. Sa, “Acousto-optics theory and application notes,” Tech. rep., AA Opto-electronic (2014).

Pixel super-resolution in optical time-stretch microscopy using acousto-optic deflector

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