15 January 1996 Real-time fluorescence microscopy monitoring of porphyrin biodistribution
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Abstract
In vivo uptake of the natural porphyrins, uroporphyrin III (UP), coproporphyrin III (CP) and protoporphyrin IX (PP), was monitored by fluorescence microscopy. Experiments were performed using the chick chorioallantoic membrane (CAM) model, which allowed video documentation of fluorescence both in real time and after integration over a chosen time interval (usually 2 s). Sensitizers at a concentration of 50 (mu) M (100 (mu) L) were injected into a medium-sized vein (diameter approximately 40 micrometer) using an ultra-fine 10 micrometer diameter needle. Fluorescence images were quantitated by subtracting the fluorescence intensity of surrounding CAM tissue (Fmatrix) from the intravascular fluorescence intensity (Fintravascular), after transformation of the video frames into digital form. The differential fluorescence intensity, Fintravascular - Fmatrix, is a measure of the biodistribution. Real time measurements clearly showed that CP and UP fluorescence is associated with moving erythrocytes and not with endothelial cells of the vessel wall. Fluorescence intensity was monitored, up to 60 minutes after injection, by averaging the fluorescence over time intervals of 2 s and recording the integrated images. The fluorescence intensity reached its maximum in about 20 - 30 min after injection, presumably after monomerization inside erythrocyte membranes. The results are interpreted in terms of physical-chemical characteristics (e.g. hydrophilicity) and correlated with the photodynamically induced hemostasis in CAM blood vessels.
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Sol Kimel, Sol Kimel, Varda Gottfried, Varda Gottfried, Karin Kunzi-Rapp, Karin Kunzi-Rapp, Nermin Akguen, Nermin Akguen, Herbert Schneckenburger, Herbert Schneckenburger, } "Real-time fluorescence microscopy monitoring of porphyrin biodistribution", Proc. SPIE 2628, Optical and Imaging Techniques for Biomonitoring, (15 January 1996); doi: 10.1117/12.229978; https://doi.org/10.1117/12.229978
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