Warren S. Grundfest

Time-resolved fluorescence spectra of arterial fluorescent compounds: Reconstruction with the laguerre expansion technique

Abstract The time-resolved fluorescence spectra of the main arterial fluorescent compounds were retrieved using a new algorithm based on the Laguerre expansion of kernels technique. Samples of elastin, collagen and cholesterol were excited with a pulsed nitrogen laser and the emission was measured at 29 discrete wavelengths between 370 and 510 nm. The expansion of the fluorescence impulse response function on the Laguerre basis of functions was optimized to reproduce the observed fluorescence emission. Collagen lifetime (5.3 ns at 390 nm) was substantially larger than that of elastin (2.3 ns) and cholesterol (1.3 ns). Two decay components were identified in the emission decay of the compounds. For collagen, the decay components were markedly wavelength dependent and hydration dependent such that the emission decay became shorter at higher emission wavelengths and with hydration. The decay characteristics of elastin and cholesterol were relatively unchanged with wavelength and with hydration. The observed variations in the time-resolved spectra of elastin, collagen and cholesterol were consistent with the existence of several fluorophores with different emission characteristics. Because the compounds are present in different proportions in healthy and atherosclerotic arterial walls, characteristic differences in their time-resolved emission spectra could be exploited to assess optically the severity of atherosclerotic lesions.

Thermal coronary angiography: a method for assessing graft patency and coronary anatomy in coronary bypass surgery

Thermal coronary angiography was evaluated in 50 patients undergoing 137 saphenous vein and 48 internal mammary artery bypass grafts. A total of 177 thermal coronary angiograms were performed after completion of the distal anastomoses by injection of cold cardioplegia into the vein or by reperfusion with warmer blood in the internal mammary artery grafts. These angiograms provided details of graft and anastomosis patency, flow directions, and presence of native coronary stenoses. Temperature differences between the injectant and the epimyocardium of greater than 4 degrees C resulted in high-contrast images. Thermal coronary angiograms were obtained in 173 of the 177 studied bypass grafts; 172 grafts were patent, and 1 internal mammary artery graft was occluded. Unsuspected stenoses were detected at the site of four distal anastomoses. Subsequently, two anastomoses were successfully revised and three additional grafts performed. Ninety-six native coronary stenoses were located in the recipient coronary arteries. In ten instances, the thermal coronary angiograms were obscured by excess fat or myocardium, thereby impeding correct image analysis. We conclude that thermal coronary angiography can be clinically relevant and helps improve decision making during coronary artery bypass operations.

Role of laser and thermal ablation devices in the treatment of vascular diseases

Since the first coronary angioplasty in 1977, both the number and complexity of interventional procedures have grown dramatically. Continuous-wave and pulsed lasers may further extend the capabilities of balloon angioplasty. Fiberoptic catheters may be used to transmit continuous-wave laser energy to ablate plaque via thermal mechanisms. Pulsed laser systems (such as the excimer) are technologically more complex than the continuous-wave systems, but may prove superior in small vessels given their ability to ablate plaque with minimal associated effects. On the other hand, modifications of the fiber-optic tip, such as the placement of a metal cap, have yielded even better results than current bare fiber systems. Such laser thermal techniques have proved a useful adjunct to balloon dilatation in peripheral vessels, but further research is necessary to determine their effect on coronary arteries. New, nonlaser technologies, however, may provide simpler power sources for thermal angioplasty. Although balloon angioplasty remains the cornerstone of interventional vascular therapy, new technologies should help to further expand the indications for nonsurgical interventions.

Characterization of type I, II, III, IV, and V collagens by time-resolved laser-induced fluorescence spectroscopy

The relative proportions of genetically distinct collagen types in connective tissues vary with tissue type and change during disease progression, development, wound healing, aging. This study aims to 1) characterize the spectro- temporal fluorescence emission of fiber different types of collagen and 2) assess the ability of time-resolved laser- induced fluorescence spectroscopy to distinguish between collagen types. Fluorescence emission of commercially available purified samples was induced with nitrogen laser excitation pulses and detected with a MCP-PMT connected to a digital storage oscilloscope. The recorded time-resolved emission spectra displayed distinct fluorescence emission characteristics for each collagen type. The time domain information complemented the spectral domain intensity data for improved discrimination between different collagen types. Our results reveal that analysis of the fluorescence emission can be used to characterize different species of collagen. Also, the results suggest that time-resolved spectroscopy can be used for monitoring of connective tissue matrix composition changes due to various pathological and non-pathological conditions.

Photobleaching of arterial fluorescent compounds: characterization of elastin, collagen and cholesterol time-resolved spectra during prolonged ultraviolet irradiation.

Abstract— To study the photobleaching of the main fluorescent compounds of the arterial wall, we repeatedly measured the time‐resolved fluorescence of elastin, collagen and cholesterol during 560 s of excitation with nitrogen laser pulses. Three fluence rate levels were used: 0.72, 7.25 and 21.75 μW/mm2. The irradiation‐related changes of the fluorescence intensity and of the time‐resolved fluorescence decay constants were characterized for the emission at 390, 430 and 470 nm. The fluorescence intensity at 390 nm decreased by 25–35% when the fluence delivered was 4 mJ/mm2 a common value in fluorescence studies of the arterial wall. Cholesterol fluorescence photobleached the most, and elastin fluorescence photobleached the least. Photobleaching was most intense at 390 nm and least intense at 470 nm such that the emission spectra of the three compounds were markedly distorted by photobleaching. The time‐resolved decay constants and the fluorescence lifetime were not altered by irradiation when the fluence was below 4 mJ/mm2. The spectral distortions associated with photobleaching complicate the interpretation of arterial wall fluorescence in terms of tissue content in elastin, collagen and cholesterol. Use of the time‐dependent features of the emission that are not altered by photobleaching should increase the accuracy of arterial wall analysis by fluorescence spectroscopy.

Time-resolved fluorescence of human aortic wall: use for improved identification of atherosclerotic lesions.

This study characterized aortic time‐resolved fluorescence spectra for stratified levels of atherosclerosis and proposed interpretation of spectrotemporal variations in terms of histologic changes.

Biliary lithotripsy as an adjunct to laparoscopic common bile duct stone extraction

SummaryWe investigated various energy sources and delivery systems suitable for fragmentation of common duct calculi by a laparoscopic technique. We evaluated electrohydraulic lithotripsy (EHL) using 1.9-Fr probe delivering 80 W and laser lithotripsy using a 200-μm fiber delivering 30–70 mJ/pulse at 5–20 Hz. In vitro biliary stone fragmentation analysis suggested that the laser lithotripsy produced a more controllable fragmentation than EHL. Initial attempts to employ EHL techniques in animal models resulted in common bile duct injury or inadequate fragmentation of stones. In contrast, biliary lithotripsy was accomplished in pigs using the pulsed-dye laser at 10 Hz and 60 mJ/pulse. Histologic evaluation revealed no evidence of ductal injury related to laser stone fragmentation. Subsequently, laser common duct lithotripsy was used in two human subjects. One patient had a 1.8-cm impacted ampullary stone and one patient had a 3-cm intrahepatic stone. In both cases, the stones were removed laparoscopically after laser fragmentation. Our experience suggests that the laser lithotripsy may facilitate laparoscopic common duct stone extraction procedures.

Consequences of scattering for spectral imaging of turbid biologic tissue

Spectral imaging permits two-dimensional mapping of the backscattering properties of biological systems. Such mapping requires broadband illumination of the entire area of interest. However, imaging of turbid biological media under these conditions often involves mean photon path lengths that exceed the pixel size. Using a numerical Monte Carlo model, we have studied the effects of photon scattering in a hemoglobin-bearing model system. We find that photon migration and the resulting wavelength-dependent optical coupling between pixels can complicate the analysis of imaging spectroscopy data. In fact, the wavelength dependence of photon trajectories also alters the distribution of photon exit angles at the tissue surface. We therefore find that the finite optical field of view of an imaging spectrometer can affect the measured spectra in the absence of chromatic aberrations.

Spectro-temporal studies of XeCl excimer laser-induced arterial wall fluorescence

We report on spectro-temporal fluorescence studies of cadaver femoral arterial walls at different stages in the progression of atherosclerosis. After excitation with a Xe-Cl excimer pulse, the time course of the fluorescence spectrum was recorded over time, and time-resolved multispectral analysis was performed. Then, under the assumption of linearity, we derived a linear spectro-temporal kernel (a weighting function) which describes the temporal behavior of the fluorescence process independently of the pulse width of the photoexcitation. The data analysis revealed both static and dynamic fluorescence characteristics which exhibited a good correlation with histological findings.

Control of excimer laser aided tissue ablation via laser-induced fluorescence monitoring

Human atherosclerotic arterial samples were ablated via a fiber with a XeCl excimer laser. The resulting tissue fluorescence was recorded for each ablating pulse. The pulse-to-pulse evolution of the fluorescence intensity at 430 nm was obtained and compared to the histological findings. The characteristic transition observed in such curves exhibited good correlation with the transition from the atheromatous layer to the normal media, as determined by the histological examination. The establishment of such a relation led to the development of a simple computer algorithm able to detect plaque to normal media transitions. The limitations of this approach are discussed.