Sensitive imaging techniques for small animals are needed to assess drug toxicity in preclinical studies. Optical coherence tomography (OCT) provides a noninvasive tool for high-resolution, depth-resolved visualization of drug-induced changes in tissue morphology. In a mouse model, we utilize OCT to assess vaginal tissue integrity following the application of topical microbicides (drugs used to prevent infection). Mice are challenged with herpes simplex virus-2 (HSV-2) to determine the correlation of tissue damage as quantified by OCT to increased susceptibility. The microbicide benzalkonium chloride (BZK) (0.02, 0.2, or 2%) or phosphate buffered saline control is administered intravaginally. In vivo OCT imaging and collection of tissue samples are performed after treatment. A quantitative OCT scoring system is applied to assess epithelial damage, and the results are compared with those of histology. A separate group of mice are treated similarly then challenged with HSV-2. Epithelial morphology quantified noninvasively by OCT and histology are dose-dependent (p<0.0001 ). The OCT scoring system detected a significant increase in epithelial damage with increasing BZK concentration (p<0.0001 ). These results paralleled an increase in HSV-2 susceptibility (p<0.005 ). OCT can be used as a noninvasive tool to assess topical drug toxicity in a small animal model with potential to predict increased susceptibility to vaginal infection.
Optoacoustic (OA) tomography has demonstrated utility in identifying blood-rich malignancies in breast tissue. We describe the development and characterization of a laser OA imaging system for the prostate (LOIS-P). The system consists of a fiber-coupled Q-switched laser operating at 757 nm, a commercial 128-channel ultrasonic probe, a digital signal processor, and software that uses the filtered radial back-projection algorithm for image reconstruction. The system is used to reconstruct OA images of a blood-rich lesion induced in vivo in a canine prostate. OA images obtained in vivo are compared to images acquired using ultrasound, the current gold standard for guiding biopsy of the prostate. Although key structural features such as the urethra could be identified with both imaging techniques, a bloody lesion representing a highly vascularized tumor could only be clearly identified in OA images. The advantages and limitations of both forward and backward illumination modes are also evaluated by collecting OA images of phantoms simulating blood vessels within tissue. System resolution is estimated to be 0.2 mm in the radial direction of the acoustic array. The minimum detectable pressure signal is 1.83 Pa. Our results encourage further development toward a dual-modality OA/ultrasonic system for prostate imaging and image-guided biopsy.
We have developed and used a laser optoacoustic imaging system with transrectal probe (LOIS-P) for detection of
mechanical lesions in canine prostates in vivo. LOIS images have been acquired with a 128-channel transrectal probe
and a 32-channel data acquisition system. Optoacoustic images showed a strong contrast enhancement for a blood
containing lesion, when compared with ultrasound images. Our studies demonstrated that sufficient optoacoustic
contrast exists between blood containing lesion and prostate tissue, although the lesion has been undetectable with
ultrasound. The imaging results have been compared with visual examination of surgically excised prostates. Although
axial resolution of the wide-band transducers employed in the transrectal probe provides good axial resolution of 0.5
mm, the convex arc geometry of the this array of transducers provides lateral resolution degrading with depth in tissue.
A two step algorithm has been developed to improve the lateral resolution of deeply located objects. This algorithm
employs optoacoustic image reconstruction based on radial
back-projection to determine location and shape of the target
object, then a procedure, we call Maximum Angular Amplitude Probability (MAAP), to determine true brightness of the
object and simultaneously remove arc-shaped artifacts associated with radial back-projection. A laser optoacoustic
imaging system (LOIS-P) with transrectal probe operating in backward detection mode empowered with the new image
reconstruction algorithm seems promising as a modality for detection of prostate cancer and guiding prostate biopsy.
Optoacoustic tomography combines advantages of pronounced optical contrast between different tissues and high resolution of ultrawide-band ultrasound imaging. Laser pulses may be effectively used to produce acoustic sources in tissue with enhanced optical absorption. Ultrasonic waves can propagate in biological tissue with minimal distortion and deliver diagnostic information to the surface of tissue, where they may be detected with temporal resolution by piezoelectric transducers. Current status of the optoacoustic tomography applied in early detection of cancerous lesions in the breast (utilizing forward mode) and in oral cavity (utilizing backward mode) is reviewed.
Oral squamous cell carcinoma is a disease which progresses through a number of well-defined morphological and biochemical changes. Optical coherence tomography (OCT) is a rapidly-evolving, non-invasive imaging modality which allows detailed probing of subsurface tissue structures with resolution on the order of microns. While this technique offers tremendous potential as a diagnostic tool for detection and characterization of oral cancer, OCT imaging is presently associated with a field of view on the order of millimeters, and acquisition time on the order of seconds. Thus, OCT's utility as a rapid cancer screening technique is presently limited. On the other hand, imaging of tissue autofluorescence provides a very rapid, high-throughput method for cancer screening. However, while autofluorescence measures may be sensitive to cancer, they are often non- specific and lead to a large number of false positives. In the present work, we have developed a fluorescence image guided optical coherence tomographic (FIG-OCT) probe in which tissue autofluorescence images are simultaneously used to guide OCT image acquisition of suspicious regions in real time. We have begun pre-clinical pilot studies with this instrument in a DMBA-induced model of oral cancer in the hamster cheek pouch. Initial results indicate that the FIG- OCT approach shows promise as a rapid and effective tool for screening of oral cancer.
Confocal opto-acoustic transducer (COAT) was developed and applied for detection of early stages of squamous cell carcinoma in hamster model of oral cancer. COAT is a novel imaging modality with optical and acoustic lens utilized for detecting in-depth opto-acoustic front surface transducer is an improved lateral resolution of 60-micrometers . The bandwidth of the confocal opto-acoustic transducer is more than 100 MHz. Therefore, in-depth axial resolution defined by the laser pulse duration and detection system equals 15-micrometers . Imaging was performed at the wavelength of the Nd:YAG laser second harmonic, which provided sufficient depth of monitoring and significant tissue contrast. Correlation of the opto- acoustic images with H and E histology sections in control animals and in animals treated with carcinogenic agent, DMBA, confirmed previous findings that early cancer lesions invisible by the naked eye may be detected with the opto- acoustic tomography. Compact design of COAT allows, in principle, application of the opto-acoustic imaging in any organ of the human digestive system.
An otoacoustic transducer capable of detecting wide-band ultrasonic profiles at the site of pulsed laser irradiation was developed and applied in imaging various stages of squamous cell carcinoma produced in a hamster cheek pouch model. Imaging was performed using 12-nanosecond Nd:YAG laser pulses (FWHM) at the wavelength of 532 nm, which provided sufficient depth of monitoring (2 mm), significant tissue contrast and 15-?m in depth resolution. The opto-acoustic images depict distinct tissue layers in control (normal) animals and heterogeneous structureless tissue in pre-cancer and cancer stages. Pronounced changes in subsurface tissue structures that occur in the course of cancer development were not detected upon gross examination. Optoacoustic images of later stages in cancer development presented exophytic or irregular profiles on the surfaces, possibly associated with enhanced microvascularization. The changes associated with benign inflammation yielded optoacoustic images different from those depicting cancer progression. The experimental results suggest that the optoacoustic images displaying layered tissue structure on the basis of variations in optical absorption and scattering may be utilized for early detection and staging of superficial mucosa cancer.
Ventricular tachycardia (VT) is a rapid and life-threatening cardiac arrhythmia that most often occurs after healing of myocardial infarction. The same catheter techniques that use resistive endocardial heating to treat supraventricular tachycardias are less effective against post-infarction VT, in part because cure of the latter arrhythmia requires large volume, deep tissue coagulation. Greater risk may also be incurred when standard percutaneous methods are used to induce deep tissue heating, because excess endocardial damage can cause mural thrombi, and a large area of non-lethal endocardial injury may itself generate VT. To address these problems we have developed a unique optical fiber for direct intramyocardial photocoagulation which, when coupled to a diode laser (805 nm), can generate lesions up to 1 cm deep and wide without disruption of the endocardium. With further refinement this system may effectively and safely cure post- infarction VT.
Christopher Frederickson, Quiang Lu, Donald Hayes, David Wallace, Michael Grove, Brent Bell, Massoud Motamedi, Sohi Rastegar, C. Wright, Charles Arcoria
Nd:YAG lasers have been used previously for selective removal of various material from teeth. To permit ablation of healthy enamel with the Nd:YAG laser, we have adopted a strategy in which micro-drops of photoabsorptive 'promoters' are placed on the enamel to enhance absorption of individual laser pulses. Ink-jet technology dispenses the micro-drops with micron- and millisecond-scale precision. Various promoters using drug and cosmetic dyes, indocyanine green, or carbon-black pigments have been studied. Typical ablation parameters are 1.064 micrometers ; 20-180 mJ per pulse; 100 microsecond(s) ; 10-30 pulses/sec; 0.2-2.0 nl drops. Recent results from the program include: (1) For a variety of promoters, a monotonic relationship obtains between absorption coefficient at 1.064 micrometers and the efficiency of ablation of enamel. (2) With different promoter volumes, the efficiency of ablation rises, plateaus, then falls with increasing volume. (3) At drilling rates of 30 pulses/sec, ablation efficiency approaches rates of 0.1 mm3/sec. LM and SEM observations show a glassy 'pebbled' crater surface indicative of hydroxyapatite that has cooled, condensed, and solidified on the crater walls. Together these results favor the view that a micro-drop promoter-assisted Nd:YAG drill can five clinically useful ablations hard dental tissue.
Thermal therapy using various heating sources such as lasers or microwaves to destroy benign and malignant lesions has recently gained widespread acceptance. However, the accurate prediction of thermal damage in tissue according to theoretical or computer modeling is difficult and unreliable due to target variability with respect to physical properties, geometry, and blood perfusion. Thus, one of the major obstacles to application of thermal therapies has been the lack of a noninvasive, real-time method that could determine the extent and geometry of treated tissue. To evaluate the effects of laser heating on tissue, we have developed an analog-digital hybrid Doppler ultrasound system to measure the phase and amplitude of ultrasonic echoes returned from the heated tissue. The system consists of an eight-gate pulsed Doppler detector, a 16-channel 12-bit A/D converter, and a signal analysis and visualization software package. In vitro studies using canine liver showed two distinct types of modulation of the echoes along the ultrasound beam path during laser irradiation using an 810 nm diode laser. Type 1 signals showed a small and slow variation in amplitude and phase, and were attributed to tissue coagulation. Type 1 signals showed a small and slow variation in amplitude and phase, and were attributed to tissue coagulation. Type 2 signals showed large and rapid variations in amplitude and phase which usually appeared after tissue surface explosion and were indicative of tissue ablation. We hypothesize that the observed phase changes in type 1 signals are due to thermal effects within the tissue consistent with tissue expansion and contraction while the phase changes in type 2 signals are likely due to formation and motion of gas bubbles in the tissue. A further development of the Doppler ultrasound technique could lead to the generation of feedback information needed for monitoring and automatic control of thermal treatment using various heating modalities such as laser, high intensity focused ultrasound, microwaves, or radio frequency waves.
Recent studies have established clinical application of laser ablation of cartilaginous tissue. The goal of this study was to investigate removal of cartilaginous tissue using diode laser. To enhance the interaction of laser light with tissue, improve the ablation efficiency and localize the extent of laser-induced thermal damage in surrounding tissue, we studied the use of a novel delivery system developed by MicroFab Technologies to dispense a known amount of Indocyanine Green (ICG) with a high spatial resolution to alter the optical properties of the tissue in a controlled fashion. Canine intervertebral disks were harvested and used within eight hours after collection. One hundred forty nL of ICG was topically applied to both annulus and nucleus at the desired location with the MicroJet prior to each irradiation. Fiber catheters (600 micrometers ) were used and positioned to irradiate the tissue with a 0.8 mm spot size. Laser powers of 3 - 10 W (Diomed, 810 nm) were used to irradiate the tissue with ten pulses (200 - 500 msec). Discs not stained with ICG were irradiated as control samples. Efficient tissue ablation (80 - 300 micrometers /pulse) was observed using ICG to enhance light absorption and confine thermal damage while there was no observable ablation in control studied. The extent of tissue damage observed microscopically was limited to 50 - 100 micrometers . The diode laser/Microjet combination showed promise for applications involving removal of cartilaginous tissue. This procedure can be performed using a low power compact diode laser, is efficient, and potentially more economical compared to procedures using conventional lasers.
Ventricular tachycardia (VT) is a rapid heart rhythm which is often life threatening. Several surgical and percutaneous ways to destroy the myocardium responsible for VT have been studied. It has been postulated that laser therapy may be ideal for this purpose because it can heat a large volume of tissue. Recent developments in diode lasers have prompted us to evaluate this source (810 nm) for photocoagulation of myocardial tissue. Its size, ease of maintenance, and cost make diode laser suitable for clinical practice in general, and for percutaneous photoablation in particular. Lesions were created in myocardium with contact irradiation using a 600 micron bare tipped optical fiber both in vitro and in vivo. Exposures of 2 to 3 W over 30 to 60 seconds created lesions with no or minimal char formations. In vivo lesions tended to be larger than in vitro, with better defined border zones. Animals tolerated laser irradiation well without significant ventricular ectopy. Diode laser irradiation is a promising means to percutaneously coagulate ventricular myocardium and for cure of VT. Further investigation of the dosimetry and healing response in both healthy and diseased myocardium is warranted.
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