Melissa L. Mather

Ultrasound techniques for characterizing colloidal dispersions

Interest in the interaction of acoustic waves with particulate mixtures has a long history—dating back to the work of Rayleigh in the 19th century. This interest has intensified over the last fifteen years as advances in electronics and instrumentation science have brought the possibility of using ultrasound to characterize colloidal mixtures both in the laboratory and in-process, and in both of these contexts a small number of instruments are currently in use. The characterization of colloidal mixtures by ultrasound requires a formal theoretical basis which relates the properties of the mixture, particularly the dispersed phase particle size distribution (PSD), to the complex wavenumber governing propagation. The number of theoretical treatments is vast, having evolved over more than a century. This paper is intended to provide a review of these developments in a form which will enable new researchers in the field to climb a very steep learning curve in a relatively short time. We discuss definitions and production techniques for colloidal mixtures and the basic physical phenomena underlying wave propagation through them. We identify two approaches to the propagation problem—scattering and coupled-phase; these are treated both separately and comparatively, particularly in relation to limitations that arise when the concentration of particles is high and the basic theories break down. We introduce the basic method for the measurement of PSD and show how dynamic effects such as flocculation and crystallization can be observed and modelled. The core of all ultrasonic characterization procedures is the physical measurement of the ultrasonic wave attenuation coefficient and phase velocity as functions of frequency; here we discuss these techniques on the basis that what is observable or measurable about a colloid depends on both its physical properties and the frequency bandwidth available for measurement. This paper concludes with our view on future developments of measurement technique and theoretical treatments.

Ultrasound evaluation of polymer gel dosimeters.

A new method for the evaluation of radiotherapy 3D polymer gel dosimeters has been developed using ultrasound to assess the significant structural changes that occur following irradiation of the dosimeters. The ultrasonic parameters of acoustic speed of propagation, attenuation and transmitted signal intensity were measured as a function of absorbed radiation dose. The dose sensitivities for each parameter were determined as 1.8 x 10(-4) s m(-1) Gy(-1), 3.9 dB m(-1) Gy(-1) and 3.2 V(-1) Gy(-1) respectively. All parameters displayed a strong variation with absorbed dose that continued beyond absorbed doses of 15 Gy. The ultrasonic measurements demonstrated a significantly larger dynamic range in dose response curves than that achieved with previously published magnetic resonance imaging (MRI) dose response data. It is concluded that ultrasound shows great potential as a technique for the evaluation of polymer gel dosimeters.

Cytoplasmic RNA in Undifferentiated Neural Stem Cells: A Potential Label-Free Raman Spectral Marker for Assessing the Undifferentiated Status

Raman microspectroscopy (rms) was used to identify, image, and quantify potential molecular markers for label-free monitoring the differentiation status of live neural stem cells (NSCs) in vitro. Label-free noninvasive techniques for characterization of NCSs in vitro are needed as they can be developed for real-time monitoring of live cells. Principal component analysis (PCA) and linear discriminant analysis (LDA) models based on Raman spectra of undifferentiated NSCs and NSC-derived glial cells enabled discrimination of NSCs with 89.4% sensitivity and 96.4% specificity. The differences between Raman spectra of NSCs and glial cells indicated that the discrimination of the NSCs was based on higher concentration of nucleic acids in NSCs. Spectral images corresponding to Raman bands assigned to nucleic acids for individual NSCs and glial cells were compared with fluorescence staining of cell nuclei and cytoplasm to show that the origin of the spectral differences were related to cytoplasmic RNA. On the basis of calibration models, the concentration of the RNA was quantified and mapped in individual cells at a resolution of ~700 nm. The spectral maps revealed cytoplasmic regions with concentrations of RNA as high as 4 mg/mL for NSCs while the RNA concentration in the cytoplasm of the glial cells was below the detection limit of our instrument (~1 mg/mL). In the light of recent reports describing the importance of the RNAs in stem cell populations, we propose that the observed high concentration of cytoplasmic RNAs in NSCs compared to glial cells is related to the repressed translation of mRNAs, higher concentrations of large noncoding RNAs in the cytoplasm as well as their lower cytoplasm volume. While this study demonstrates the potential of using rms for label-free assessment of live NSCs in vitro, further studies are required to establish the exact origin of the increased contribution of the cytoplasmic RNA.

Ultrasound tomography imaging of radiation dose distributions in polymer gel dosimeters: Preliminary study

A novel imaging system for investigation of absorbed dose distributions in radiotherapy polymer gel dosimeters using ultrasound is introduced. A prototype transmission ultrasound computed tomography (UCT) imaging system is developed and evaluated. The imaging capabilities of the system are assessed through investigation of an irradiated polyacrylamide gel test phantom. Images based on transmitted signal amplitude and time of flight (TOF) of the ultrasonic signal through the phantom are reconstructed using a filtered backprojection technique. In general, the reconstruction of the square field in the TOF image was superior to the transmission image, however, transmission images displayed superior contrast to TOF images. The image quality achieved with this prototype system is promising and could be significantly enhanced through improvements, in particular through the development of more sophisticated experimental equipment. It is concluded that UCT is a viable technique for imaging absorbed dose distributions in polymer gel dosimeters and investigations are continuing to further improve the system.

Investigation of ultrasonic properties of PAG and MAGIC polymer gel dosimeters

Ultrasonic speed of propagation and attenuation were investigated as a function of absorbed radiation dose in PAG and MAGIC polymer gel dosimeters. Both PAG and MAGIC gel dosimeters displayed a dependence of ultrasonic parameters on absorbed dose with attenuation displaying significant changes in the dose range investigated. The ultrasonic attenuation dose sensitivity at 4 MHz in MAGIC gels was determined to be 4.7 +/- 0.3 dB m(-1) Gy(-1) and for PAG 3.9 +/- 0.3 dB m(-1) Gy(-1). Ultrasonic speed dose sensitivities were 0.178 +/- 0.006 m s(-1) Gy(-1) for MAGIC gel and -0.44 +/- 0.02 m s(-1) Gy(-1) for PAG. Density and compressional elastic modulus were investigated to explain the different sensitivities of ultrasonic speed to radiation for PAG and MAGIC gels. The different sensitivities were found to be due to differences in the compressional elastic modulus as a function of dose for the two formulations. To understand the physical phenomena underlying the increase in ultrasonic attenuation with dose, the viscoelastic properties of the gels were studied. Results suggest that at ultrasonic frequencies, attenuation in polymer gel dosimeters is primarily due to volume viscosity. It is concluded that ultrasonic attenuation significantly increases with absorbed dose. Also, the ultrasonic speed in polymer gel dosimeters is affected by changes in dosimeter elastic modulus that are likely to be a result of polymerization. It is suggested that ultrasound is a sufficiently sensitive technique for polymer gel dosimetry.

Image-based characterization of foamed polymeric tissue scaffolds

Tissue scaffolds are integral to many regenerative medicine therapies, providing suitable environments for tissue regeneration. In order to assess their suitability, methods to routinely and reproducibly characterize scaffolds are needed. Scaffold structures are typically complex, and thus their characterization is far from trivial. The work presented in this paper is centred on the application of the principles of scaffold characterization outlined in guidelines developed by ASTM International. Specifically, this work demonstrates the capabilities of different imaging modalities and analysis techniques used to characterize scaffolds fabricated from poly(lactic-co-glycolic acid) using supercritical carbon dioxide. Three structurally different scaffolds were used. The scaffolds were imaged using: scanning electron microscopy, micro x-ray computed tomography, magnetic resonance imaging and terahertz pulsed imaging. In each case two-dimensional images were obtained from which scaffold properties were determined using image processing. The findings of this work highlight how the chosen imaging modality and image-processing technique can influence the results of scaffold characterization. It is concluded that in order to obtain useful results from image-based scaffold characterization, an imaging methodology providing sufficient contrast and resolution must be used along with robust image segmentation methods to allow intercomparison of results.

Longitudinal acoustic properties of poly(lactic acid) and poly(lactic-co-glycolic acid)

Acoustics offers rich possibilities for characterizing and monitoring the biopolymer structures being employed in the field of biomedical engineering. Here we explore the rudimentary acoustic properties of two common biodegradable polymers: poly(lactic acid) and poly(lactic-co-glycolic acid). A pulse-echo technique is developed to reveal the bulk speed of sound, acoustic impedance and acoustic attenuation of small samples of the polymer across a pertinent temperature range of 0-70 °C. The glass transition appears markedly as both a discontinuity in the first derivative of the speed of sound and a sharp increase in the acoustic attenuation. We further extend our analysis to consider the role of ethanol, whose presence is observed to dramatically modify the acoustic properties and reduce the glass transition temperature of the polymers. Our results highlight the sensitivity of acoustic properties to a range of bulk properties, including visco-elasticity, molecular weight, co-polymer ratio, crystallinity and the presence of plasticizers.

Meeting the needs of monitoring in tissue engineering

Tissue engineering is a rapidly growing field that aims to develop biological substitutes that restore, maintain or improve tissue function. The focus of research to date has been the underlying biology required for tissue-engineered therapies. However, as tissue-engineered products reach the marketplace, there is a pressing need for an improved understanding of the engineering and economic issues associated with them. This is motivated by the lack of commercial viability of many of the initial therapies that have been produced. It has been suggested in the literature that this is partly due to poor process and system design in tissue production, as well as a lack of process monitoring and control. This review argues that principles of design, measurement and process monitoring from the physical sciences are needed to move tissue engineering forward, and that much of the technology needed to realize this is already available.

Measurement of ultrasonic attenuation coefficient in polymer gel dosimeters.

A technique is described for investigation of the ultrasonic attenuation coefficient for evaluation of absorbed dose in polymer gel dosimeters. Using this technique the attenuation coefficient as a function of absorbed dose in PAG and MAGIC polymer gel dosimeters was measured. The ultrasonic attenuation coefficient dose sensitivity for PAG was found to be 2.9 +/- 0.3 dB m(-1) Gy(-1) and for MAGIC gel 4.2 +/- 0.3 dB m(-1) Gy(-1). Unlike previous studies of ultrasonic attenuation in polymer gel dosimeters this technique enables a direct measure of the attenuation coefficient.

Ultrasonic absorption in polymer gel dosimeters.

Ultrasonic absorption in polymer gel dosimeters was investigated. An ultrasonic interferometer was used to study the frequency (f) dependence of the absorption coefficient (alpha) in a polyacrylamide gel dosimeter (PAG) in the frequency range 5-20 MHz. The frequency dependence of ultrasonic absorption deviated from that of an ideal viscous fluid. The presence of relaxation mechanisms was evidenced by the frequency dependence of alpha/f(2) and the dispersion in ultrasonic velocity. It was concluded that absorption in polymer gel dosimeters is due to a number of relaxation processes which may include polymer-solvent interactions as well as relaxation due to motion of polymer side groups. The dependence of ultrasonic absorption on absorbed dose and formulation was also investigated in polymer gel dosimeters as a function of pH and chemical composition. Changes in dosimeter pH and chemical composition resulted in a variation in ultrasonic dose response curves. The observed dependence on pH was considered to be due to pH induced modifications in the radiation yield while changes in chemical composition resulted in differences in polymerisation kinetics.