Acoustic resolution photoacoustic Doppler velocimetry in blood-mimicking fluids

Photoacoustic Doppler velocimetry provides a major opportunity to overcome limitations of existing blood flow measuring methods. By enabling measurements with high spatial resolution several millimetres deep in tissue, it could probe microvascular blood flow abnormalities characteristic of many different diseases. Although previous work has demonstrated feasibility in solid phantoms, measurements in blood have proved significantly more challenging. This difficulty is commonly attributed to the requirement that the absorber spatial distribution is heterogeneous relative to the minimum detectable acoustic wavelength. By undertaking a rigorous study using blood-mimicking fluid suspensions of 3 μm absorbing microspheres, it was discovered that the perceived heterogeneity is not only limited by the intrinsic detector bandwidth; in addition, bandlimiting due to spatial averaging within the detector field-of-view also reduces perceived heterogeneity and compromises velocity measurement accuracy. These detrimental effects were found to be mitigated by high-pass filtering to select photoacoustic signal components associated with high heterogeneity. Measurement under-reading due to limited light penetration into the flow vessel was also observed. Accurate average velocity measurements were recovered using "range-gating", which furthermore maps the cross-sectional velocity profile. These insights may help pave the way to deep-tissue non-invasive mapping of microvascular blood flow using photoacoustic methods

Velocity measurements in whole blood using acoustic resolution photoacoustic Doppler

Acoustic resolution photoacoustic Doppler velocimetry promises to overcome the spatial resolution and depth penetration limitations of current blood flow measuring methods. Despite successful implementation using blood-mimicking fluids, measurements in blood have proved challenging, thus preventing in vivo application. A common explanation for this difficulty is that whole blood is insufficiently heterogeneous relative to detector frequencies of tens of MHz compatible with deep tissue photoacoustic measurements. Through rigorous experimental measurements we provide new insight that refutes this assertion. We show for the first time that, by careful choice of the detector frequency and field-of-view, and by employing novel signal processing methods, it is possible to make velocity measurements in whole blood using transducers with frequencies in the tens of MHz range. These findings have important implications for the prospects of making deep tissue measurements of blood flow relevant to the study of microcirculatory abnormalities associated with cancer, diabetes, atherosclerosis and other conditions

Development of a blood oxygenation phantom for photoacoustic tomography combined with online pO2 detection and flow spectrometry

Photoacoustic tomography (PAT) is intrinsically sensitive to blood oxygen saturation (sO2) in vivo. However, making accurate sO2 measurements without knowledge of tissue- and instrumentation-related correction factors is extremely challenging. We have developed a low-cost flow phantom to facilitate validation of PAT systems. The phantom is composed of a flow circuit of tubing partially embedded within a tissue-mimicking material, with independent sensors providing online monitoring of the optical absorption spectrum and partial pressure of oxygen in the tube. We first test the flow phantom using two small molecule dyes that are frequently used for photoacoustic imaging: methylene blue and indocyanine green. We then demonstrate the potential of the phantom for evaluating sO2 using chemical oxygenation and deoxygenation of blood in the circuit. Using this dynamic assessment of the photoacoustic sO2 measurement in phantoms in relation to a ground truth, we explore the influence of multispectral processing and spectral coloring on accurate assessment of sO2. Future studies could exploit this low-cost dynamic flow phantom to validate fluence correction algorithms and explore additional blood parameters such as pH and also absorptive and other properties of different fluids

Photoacoustic Doppler velocity measurements using time-domain cross-correlation

The feasibility of making spatially resolved measurements of blood velocity using a pulsed photoacoustic Doppler technique has been investigated. Doppler time shifts were quantified via cross-correlation of photoacoustic waveform pairs. The waveforms were generated within a blood-simulating phantom using pairs of light pulses and detected using an ultrasound transducer. Two types of blood-simulating phantom were investigated. The first was a rotating wheel phantom consisting of micron-scale absorbers imprinted on an acetate sheet and moved at known velocities; this simulated plug flow. A time-correlation data processing scheme was used to quantify velocities in the range 0.15 to 1.5 m/s with accuracies as low as 1% and a measurement resolution <4%. The transducer beam width determines a maximum measurable velocity |Vmax| beyond which correlation is lost due to absorbers moving out of the focal beam between the two laser pulses. Resolution and |Vmax| can be scaled to much lower velocities such as those encountered in microvasculature (< 50 mm/s). Velocities in this range were investigated for the second type of phantom comprising absorbers, such as red blood cells or microspheres, flowing in a suspension within a transparent tube; this demonstrated non-plug flow. The absorber-filled tube could also be manually shifted for direct comparison between the plug and non-plug flow cases. Laminar flow gave rise to under-reading of the known velocities, which was exacerbated by increasing absorber concentrations and tube diameters, presumably due to inadequate light penetration into the tube. A novel signal processing scheme (“waveform segmentation”) was developed to surmount this difficulty, and also adds the potential for mapping out the flow velocity profile across the tube. The results show that the absorber spatial heterogeneity can be resolved even using a relatively low frequency detector, and thus pave the way for applying the cross-correlation technique to make blood velocity measurements in vivo

Evaluation of precision in optoacoustic tomography for preclinical imaging in living subjects.

Optoacoustic Tomography (OT) is now widely used in preclinical imaging, however, precision (repeatability and reproducibility) of OT has yet to be determined. METHODS: We used a commercial small animal OT system. Measurements in stable phantoms were used to independently assess the impact of system variables on precision (using coefficient of variation, COV), including acquisition wavelength, rotational position, frame averaging. Variables due to animal handling and physiology, such as anatomical placement and anesthesia conditions were then assessed in healthy nude mice using the left kidney and spleen as reference organs. Temporal variation was assessed by repeated measurements over hours and days both in phantoms and $\textit{in vivo}$. Sensitivity to small molecule dyes was determined in phantoms and $\textit{in vivo}$; precision was assessed $\textit{in vivo}$ using IRDye800CW. RESULTS: OT COV in a stable phantom was less than 2% across all wavelengths over 30 days. The factors with greatest impact on the signal repeatability in phantoms were rotational position and user experience, both of which still resulted in a COV of less than 4%. Anatomical ROI size showed the highest variation at 12% and 18% COV in the kidney and spleen respectively, however, functional SO₂ measurements based on a standard operating procedure showed exceptional reproducibility of <4% COV. COV for repeated injections of IRDye800CW was 6.6%. Sources of variability for $\textit{in vivo}$ data included respiration rate, user experience and animal placement. CONCLUSION: Data acquired with our small animal OT system was highly repeatable and reproducible across subjects and over time. Therefore, longitudinal OT studies may be performed with high confidence when our standard operating procedure is followed.This work was funded by: the EPSRC-CRUK Cancer Imaging Centre in Cambridge and Manchester (C197/A16465); CRUK (C14303/A17197, C47594/A16267); EU-FP7-agreement FP7-PEOPLE-2013-CIG-630729; and the University of Cambridge EPSRC Impact Acceleration Account

The Star Formation Across Cosmic Time (SFACT) Survey. III. Spectroscopy of the Initial Catalog of Emission-Line Objects

The Star Formation Across Cosmic Time (SFACT) survey is a new narrowband survey designed to detect emission-line galaxies (ELGs) and quasi-stellar objects (QSOs) over a wide range of redshifts in discrete redshift windows. The survey utilizes the WIYN 3.5m telescope and the Hydra multifiber positioner to perform efficient follow-up spectroscopy on galaxies identified in the imaging part of the survey. Since the objects in the SFACT survey are selected by their strong emission lines, it is possible to obtain useful spectra for even the faintest of our sources (r ~ 25). Here we present the 453 objects that have spectroscopic data from the three SFACT pilot-study fields, 415 of which are confirmed ELGs. The methodology for processing and measuring these data is outlined in this paper and example spectra are displayed for each of the three primary emission lines used to detect objects in the survey (H-alpha, [O III]5007, and [O II]3727). Spectra of additional QSOs and non-primary emission-line detections are also shown as examples. The redshift distribution of the pilot-study sample is examined and the ELGs are placed in different emission-line diagnostic diagrams in order to distinguish the star-forming galaxies from the active galactic nuclei.Comment: 20 pages, 10 figures. Accepted for publication in the Astronomical Journa

Properties of the KISS Green Pea Galaxies

Green peas (GPs) are a class of extreme star-forming galaxies (SFGs) at intermediate redshifts, originally discovered via color selection using multifilter, wide-field survey imaging data. They are commonly thought of as being analogs of high-redshift Lyα-emitting galaxies. The defining characteristic of GP galaxies is a high-excitation nebular spectrum with very large equivalent width lines, leading to the recognition that GP-like galaxies can also be identified in samples of emission-line galaxies. Here we compare the properties a sample of [O iii]-selected SFGs (z = 0.29–0.41) from the KPNO International Spectroscopic Survey (KISS) with the color-selected GPs. We find that the KISS [O iii]-selected galaxies overlap with the parameter space defined by the color-selected GPs; the two samples appear to be drawn from the same population of objects. We compare the KISS GPs with the full Hα-selected KISS SFG sample (z < 0.1) and find that they are extreme systems. Many appear to be young systems at their observed look-back times (3–4 Gyr), with more than 90% of their rest-frame B-band luminosity coming from the starburst population. We compute the volume density of the KISS red (KISSR) GPs at z = 0.29–0.41 and find that they are extremely rare objects. We do not see galaxies as extreme as the KISSR GPs in the local universe, although we recognize several lower-luminosity systems at z < 0.1

Photoacoustic imaging using genetically encoded reporters: a review

Genetically encoded contrast in photoacoustic imaging (PAI) is complementary to the intrinsic contrast provided by endogenous absorbing chromophores such as hemoglobin. The use of reporter genes expressing absorbing proteins opens the possibility of visualizing dynamic cellular and molecular processes. This is an enticing prospect but brings with it challenges and limitations associated with generating and detecting different types of reporters. The purpose of this review is to compare existing PAI reporters and signal detection strategies, thereby offering a practical guide, particularly for the nonbiologist, to choosing the most appropriate reporter for maximum sensitivity in the biological and technological system of interest.J.B. and S.E.B. are supported by the EPSRCCRUK Cancer Imaging Centre in Cambridge and Manchester (No. C197/A16465); CRUK (Nos. C14303/A17197 and C47594/A16267); and the European Union’s Seventh Framework Programme (No. FP7/2007-2013) under Grant Agreement No. FP7-PEOPLE-2013-CIG-630729. J.Y. is partly supported by Duke MEDx Basic Research Grant. J.L. acknowledges the support of ERC Starting Grant No. 281356

Properties of the KISS Green Pea Galaxies

Green Peas are a class of extreme star-forming galaxies at intermediate redshifts, originally discovered via color-selection using multi-filter, wide-field survey imaging data (Cardamone et al. 2009). They are commonly thought of as being analogs of high-redshift Ly$\alpha$-emitting galaxies. The defining characteristic of Green Pea galaxies is a high-excitation nebular spectrum with very large equivalent width lines, leading to the recognition that Green Pea-like galaxies can also be identified in samples of emission-line galaxies. Here we compare the properties a sample of [O III]-selected star-forming galaxies (z = 0.29-0.41) from the KPNO International Spectroscopic Survey (KISS) with the color-selected Green Peas. We find that the KISS [O III]-selected galaxies overlap with the parameter space defined by the color-selected Green Peas; the two samples appear to be drawn from the same population of objects. We compare the KISS Green Peas with the full H$\alpha$-selected KISS star-forming galaxy sample (z $<$ 0.1) and find that they are extreme systems. Many appear to be young systems at their observed look-back times (3-4 Gyr), with more than 90% of their rest-frame B-band luminosity coming from the starburst population. We compute the volume density of the KISSR Green Peas at z = 0.29-0.41 and find that they are extremely rare objects. We don't see galaxies as extreme as the KISSR Green Peas in the local Universe, although we recognize several lower-luminosity systems at z $<$ 0.1.Comment: 21 pages, 12 figures. Accepted for publication in the Astrophysical Journa