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Microphotometric Techniques In Intravital Microcirculatory Studies
Author(s) -
Witte Siegfried
Publication year - 1979
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1979.tb00222.x
Subject(s) - absorbance , fluorescence , chemistry , intravital microscopy , biomedical engineering , blood capillary , fluorescein , biophysics , capillary action , analytical chemistry (journal) , microcirculation , materials science , chromatography , optics , biology , circulatory system , physics , medicine , composite material , radiology , endocrinology
SUMMARY Intravital microscopic techniques were developed in order to measure plasma proteins: (1) circulating in the streaming blood of microcirculatory vessels of the exposed rat mesentery, (2) permeating the vessel wall, and (3) moving through the perivascular interstitial tissue. By means of the ultramicrospectrophotometry, the absorbance of proteins containing aromatic amino acids at 280 nm is measured. Absolute amounts of proteins in the range of picograms are measured in a circular measuring field with a diameter of 17 μm. Problems of calibration and of un‐specific light scattering are discussed. The scanning microfluorometry makes use of the fluorescent labelling of the plasma proteins. Interstitial tissue with a capillary vessel containing labelled proteins is measured at high speed. A motor‐driven object stage synchronized with a measuring system uses scanning lines to traverse the tissue. The data are then processed by a computer. In this way we get continuous quantitative information about the dynamics of protein permeability, starting from time zero. The measuring of the fluorescence intensity and the speed of the fluorometric system are further improved by a computerized 10 kHz scanning device.