Due to the wide use of animals for human disease studies, small animal whole-body imaging plays an increasingly
important role in biomedical research. Currently, none of the existing imaging modalities can provide both anatomical
and glucose metabolic information, leading to higher costs of building dual-modality systems. Even with image coregistration,
the spatial resolution of the metabolic imaging modality is not improved. We present a ring-shaped confocal
photoacoustic computed tomography (RC-PACT) system that can provide both assessments in a single modality.
Utilizing the novel design of confocal full-ring light delivery and ultrasound transducer array detection, RC-PACT
provides full-view cross-sectional imaging with high spatial resolution. Scanning along the orthogonal direction provides
three-dimensional imaging. While the mouse anatomy was imaged with endogenous hemoglobin contrast, the glucose
metabolism was imaged with a near-infrared dye-labeled 2-deoxyglucose. Through mouse tumor models, we
demonstrate that RC-PACT may be a paradigm shifting imaging method for preclinical research.
With the increasing use of small animals for human disease studies, small-animal whole-body molecular imaging plays an important role in biomedical research. Currently, none of the existing imaging modalities can provide both anatomical and glucose molecular information, leading to higher costs of building dual-modality systems. Even with image co-registration, the spatial resolution of the molecular imaging modality is not improved. Utilizing a ring-shaped confocal photoacoustic computed tomography system, we demonstrate, for the first time, that both anatomy and glucose uptake can be imaged in a single modality. Anatomy was imaged with the endogenous hemoglobin contrast, and glucose metabolism was imaged with a near-infrared dye-labeled 2-deoxyglucose.
We report a novel small-animal whole-body imaging system called ring-shaped confocal photoacoustic computed tomography (RC-PACT). RC-PACT is based on a confocal design of free-space ring-shaped light illumination and 512-element full-ring ultrasonic array signal detection. The free-space light illumination maximizes the light delivery efficiency, and the full-ring signal detection ensures a full two-dimensional view aperture for accurate image reconstruction. Using cylindrically focused array elements, RC-PACT can image a thin cross section with 0.10 to 0.25 mm in-plane resolutions and 1.6 s/frame acquisition time. By translating the mouse along the elevational direction, RC-PACT provides a series of cross-sectional images of the brain, liver, kidneys, and bladder.
pH is a tightly regulated indicator of metabolic activity. In mammalian systems, imbalance of pH regulation may result
from or result in serious illness. Even though the regulation system of pH is very robust, tissue pH can be altered in many
diseases such as cancer, osteoporosis and diabetes mellitus. Traditional high-resolution optical imaging techniques, such
as confocal microscopy, routinely image pH in cells and tissues using pH sensitive fluorescent dyes, which change their
fluorescence properties with the surrounding pH. Since strong optical scattering in biological tissue blurs images at
greater depths, high-resolution pH imaging is limited to penetration depths of 1mm. Here, we report photoacoustic
microscopy (PAM) of commercially available pH-sensitive fluorescent dye in tissue phantoms. Using both opticalresolution
photoacoustic microscopy (OR-PAM), and acoustic resolution photoacoustic microscopy (AR-PAM), we
explored the possibility of recovering the pH values in tissue phantoms. In this paper, we demonstrate that PAM was
capable of recovering pH values up to a depth of 2 mm, greater than possible with other forms of optical microscopy.
KEYWORDS: Transducers, Photoacoustic tomography, Whole body imaging, Imaging systems, Data acquisition, Ultrasonics, In vivo imaging, Photoacoustic spectroscopy, 3D image reconstruction, Confocal microscopy
We report herein a novel three-dimensional photoacoustic computed tomography (PACT) system for small-animal
whole-body imaging. The PACT system, based on a 512-element
full-ring ultrasonic transducer array, was cylindrically
focused and capable of forming a two-dimensional image in 1.6 seconds. The pulsed laser could either illuminate
directly from the top or be reshaped to illuminate the sample from the side. Top illumination was mainly used for mouse
brain and mouse embryo imaging. Side illumination provided in vivo anatomical images of an adult mouse. By
translating the mouse along the elevational direction, the system provided serial cross-sectional images.
pH is a tightly regulated indicator of metabolic activity. In mammalian systems, an imbalance of pH regulation may result from or result in serious illness. In this paper, we report photoacoustic microscopy (PAM) of a commercially available pH-sensitive fluorescent dye (SNARF-5F carboxylic acid) in tissue phantoms. We demonstrated that PAM is capable of pH imaging in absolute values at tissue depths of up to 2.0 mm, greater than possible with other forms of optical microscopy.
Traditionally micro-well plate based platforms used in biology utilize fluorescence intensity based methods to measure
processes of biological relevance. However, fluorescence intensity measurements suffer from calibration drift due to a
variety of factors. Photobleaching and self-quenching of the fluorescent dyes cause the intensity signal to drop over the
lifetime of sensor immobilized inside the well. Variation in turbidity of the sample during the course of the measurement
affects the measured fluorescence intensity. In comparison, fluorescence lifetime measurements are not significantly
affected by these factors because fluorescence lifetime is a physico-chemical property of the fluorescent dye. Reliable
and inexpensive frequency domain fluorescence lifetime instrumentation platforms are possible because the greater
tolerance for optical alignment, and because they can be performed using inexpensive light sources such as LEDs. In this
paper we report the development of a frequency domain fluorescence lifetime well-plate platform utilizing an oxygen
sensitive transition-metal ligand complex fluorophore with a lifetime in the microsecond range. The fluorescence
lifetime dye is incorporated in a polymer matrix and immobilized on the base of micro-well of a 60 well micro-well
plate. Respiration measurements are performed in both aqueous and non-aqueous environment. Respirometry
measurements were recorded from single Daphnia magna egg in hard water. Daphnia is an aquatic organism, important
in environmental toxicology as a standard bioassay and early warning indicator for water quality monitoring. Also
respirometry measurements were recorded from Tribolium castaneum eggs, which are common pests in the processed
flour industry. These eggs were subjected to mitochondrial electron transport chain inhibitor such as potassium cyanide
(KCN) and its effects on egg respiration were measured in real-time.
The detection of harmful chemicals and biological agents in real time is a critical need for protecting water quality. We
studied the real-time effects of five environmental contaminants with differing modes of action (atrazine,
pentachlorophenol, cadmium chloride, malathion, and potassium cyanide) on respiratory oxygen consumption in 2-day
post-fertilization fathead minnow (Pimephales promelas) eggs. Our objective was to assess the sensitivity of fathead
minnow eggs using the self-referencing micro-optrode technique to detect instantaneous changes in oxygen consumption
after brief exposures to low concentrations of contaminants. Oxygen consumption data indicated that the technique is
indeed sensitive enough to reliably detect physiological alterations induced by all contaminants. After 2 h of exposure,
we identified significant increases in oxygen consumption upon exposure to pentachlorophenol (100 and 1000 μg/L),
cadmium chloride (0.0002 and 0.002 μg/L), and atrazine (150 μg/L). In contrast, we observed a significant decrease in
oxygen flux after exposures to potassium cyanide (5.2, 22, and 44 μg/L) and atrazine (1500 μg/L). No effects were
detected after exposures to malathion (200 and 340 μg/L). We have also tested the sensitivity of Daphnia magna
embryos as another animal model for real-time environmental biomonitoring. Our results are so far encouraging and
support further development of this technology as a physiologically coupled biomonitoring tool for the detection of
environmental toxicants.
The reference optical path is essential for optical systems which function on the basis of light interference. In the case of frequency domain (FD) fluorescence life-time optrodes, a reference LED is used as a standard for calculating the phase angle. The reference LED is configured so that radiation travels the same length to the detector as that of the fluorescence signal being analyzed. The phase shift, which provides details of fluorescence lifetime, is measured between these two signals - the fluorescence signal and reference LED signal, using a photodetector. We have designed, developed and implemented a FD optrode system without a reference LED. The key requirement of such a system is that phase shifts due to optics at wavelength of fluorescence and electronics have to be calibrated. In the reference-free system, the reference signal comes from the lock-in-amplifier which also drives the excitation LED. The lock-in-amplifier measures the phase shift between the excitation signal and the fluorescence emission signal from the photodetector and is locked at the frequency of modulation of the excitation signal. This insures higher signal to noise ratio and low-noise measurements. The reference-free optrode system removes some constraints on the coupling optics, which help improve the overall performance of the system. After development of electronics, and optimization of coupling optics, the system was calibrated in different oxygen concentration solutions to measure fluorescence intensity and lifetime of the oxygen sensitive dye platinum tetrakis (pentafluorophenyl) porphine (PtTFPP).
Fiber optic based sensor technologies have many significant advantages over electrochemical sensors, and as a result
have broad application for sensing in biology, agriculture and medicine. An important component of fiber optic
biosensor is the sensing element. Usually, a polymer matrix containing the analyte specific fluorescent dye is
immobilized on one end of the fiber optic probe. The polymer matrix provides mechanical stability to the immobilized
membrane and entraps the fluorescent dye molecules. The target analyte diffuses into the membrane polymer and
quenches the fluorescent dye. This optical interaction between the analyte and fluorescent dye dynamically changes the
fluorescence lifetime of the dye. These changing fluorescent lifetimes reveal information about the target analyte
concentration. Although the fluorescent lifetime of the dye is of primary interest, high signal-to-noise ratio (SNR) is also
very desirable. Conventionally, complex electronics is implemented to achieve high SNR. Various signal processing
methods such as signal amplification and filtering are implemented to achieve high SNR. In this study, we report optical
signal amplification by modification of the dye-polymer matrix by addition of titanium oxide particles. The addition of
titanium oxide particles enhanced the optical signal intensity. Optical performances of different sizes and concentrations
of titanium oxide particles are compared. We believe that this increased optical intensity is due to increased optical
scattering in the dye-polymer matrix. We also compare the performance of titanium oxide particles to gold and other
material particles to experimentally probe the exact nature of light scattering in the dye-polymer matrix.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.