Distribution Of Fluorescent Dye Research Articles

Assessment of Hyperosmolar Blood-Brain Barrier Opening in Glioblastoma via Histology with Evans Blue and DCE-MRI.

While the blood-brain barrier (BBB) is often compromised in glioblastoma (GB), the perfusion and consequent delivery of drugs are highly heterogeneous. Moreover, the accessibility of drugs is largely impaired in the margins of the tumor and for infiltrating cells at the origin of tumor recurrence. In this work, we evaluate the value of methods to assess hemodynamic changes induced by a hyperosmolar shock in the core and the margins of a tumor in a GB model. Osmotic shock was induced with an intracarotid infusion of a hypertonic solution of mannitol in mice grafted with U87-MG cells. The distribution of fluorescent dye (Evans blue) within the brain was assessed via histology. Dynamic contrast-enhanced (DCE)-MRI with an injection of Gadolinium-DOTA as the contrast agent was also used to evaluate the effect on hemodynamic parameters and the diffusion of the contrast agent outside of the tumor area. The histological study revealed that the fluorescent dye diffused much more largely outside of the tumor area after osmotic shock than in control tumors. However, the study of tumor hemodynamic parameters via DCE-MRI did not reveal any change in the permeability of the BBB, whatever the studied MRI parameter. The use of hypertonic mannitol infusion seems to be a promising method to increase the delivery of compounds in the margins of GB. Nevertheless, the DCE-MRI analysis method using gadolinium-DOTA as a contrast agent seems of limited value for determining the efficacy of opening the BBB in GB after osmotic shock.

Effect of Ion Motion on Zeta-Potential Distribution at Microchannel Wall Obtained from Nanoscale Laser-Induced Fluorescence

The present study has experimentally investigated the two-dimensional distribution of zeta-potential at the wall, which dominates electroosmotic microchannel flow. Nanoscale laser-induced fluorescence imaging using fluorescent dye and the evanescent wave with total internal reflection was developed for the zeta-potential measurement. The fluorescent dye in the vicinity of the wall is excited by the evanescent wave, which decays exponentially from the wall. The zeta-potential is obtained from the fluorescent intensity because the distribution of fluorescent dye near the wall is related to the zeta-potential by the Boltzman distribution. Two kinds of solution at different Na+ concentrations were mixed in a T-shaped microchannel composed of PDMS and silica glass. The zeta-potential distribution at the silica glass wall was measured with the uncertainty of 4.7 mV. The motion of Na+ in the microchannel was estimated by the numerical analysis using the velocity information obtained by micrometer-resolution particle image velocimetry. It is concluded that electroosmotic flow was generated by the zeta-potential distribution at the silica glass and PDMS wall, which was dependent on the Na+ transport in the flow field.

Pure‐Phase Transport and Dissolution of TCE in Sedimentary Rock Saprolite

The objective of this study was to experimentally determine the influence of pore structure on the transport and dissolution of trichloroethylene (TCE) in clay-rich saprolite. In order to simulate a "spill," pure-phase TCE containing a water-insoluble fluorescent dye was injected into two heterogeneous 24-cm-diameter by 37-cm-long undisturbed columns of water-saturated saprolite. TCE entry occurred at capillary pressures of 2.7 and 4.0 kPa. Ten or 28 d after injection, the column was sliced horizontally into three sections and visually examined. The distribution of fluorescent dye indicated that pure-phase TCE migrated mainly through fractures in the shale saprolite and through fine root holes or other macropores in the limestone saprolite residuum. Analysis of saprolite subsamples indicated that TCE was present throughout much of the saprolite column but usually at concentrations less than the solubility of TCE. This spreading was caused by diffusion, which also contributed to the rapid dissolution of TCE in the fractures and macropores. Modeling was carried out using previously published dissolution and diffusion equations. The calculations confirm that rapid disappearance of immiscible TCE can occur in this type of material because of the small size of fracture or macropore openings and the high porosity of the fine-grained material. This study indicates that industrial solvents can readily enter fractures and macropores in otherwise very fine-grained subsoils and then rapidly dissolve and diffuse into the fine-pore structure, from which they may be very difficult to remove.

Fringe-scan flow cytometry.

We describe the development of a scanning flow cytometer capable of measuring the distribution of fluorescent dye along objects with a spatial resolution of 0.7 micron. The heart of this instrument, called a fringe-scan flow cytometer, is an interference field (i.e., a series of intense planes of illumination) produced by the intersection of two laser beams. Fluorescence profiles (i.e., records showing the intensity of fluorescence measured at 20 ns intervals) are recorded during the passage of objects through the fringe field. The shape of the fringe field is determined by recording light scatter profiles as 0.25 micron diameter microspheres traverse the field. The distribution of the fluorescent dye along each object passing through the fringe field is estimated from the recorded fluorescence profile using Fourier deconvolution. We show that the distribution of fluorescent dye along microsphere doublets and along propidium iodide stained human chromosomes can be determined accurately using fringe-scan flow cytometry. The accuracy of fringe-scan shape analysis was determined by comparing fluorescence profiles estimated from fringe-scan profiles for microspheres and chromosomes with fluorescence profiles for the same objects measured using slit-scan flow cytometry.

Function of the Precapillary Vessels in Peripheral Vascular Disease

This article reviews the present knowledge of vasomotion in skin capillaries of patients with peripheral vascular disease, in particular Raynaud's disease and acrocyanosis. The data are based on measurements of red blood-cell velocity and of fluorescent-dye distribution in capillary areas obtained by videomicroscopy in combination with videodensitometric evaluation. Normally, adjacent skin capillaries exhibit similar flow patterns and are filled at the same time after i.v. Na-fluorescein injection, whereas in acrocyanosis, pathological vasomotion occurs frequently (different flow patterns in adjacent capillaries, delayed filling of single capillaries or capillary groups). Inhomogeneous perfusion is also common in microangiopathy caused by scleroderma. Hypersensitivity to a standardized cold stimulus is the main feature of primary Raynaud's disease.

Characterisation of hepatocyte sub-populations generated by centrifugal elutriation.

We describe protocols for the fractionation of isolated hepatocytes into eight sub-populations using centrifugal elutriation. The distribution of fluorescein isothiocyanate and acridine orange in hepatocytes prepared from livers pre-perfused with one of these dyes is described and used as an indicator of acinar zone derivation for each population. The cytochrome P-450 content and response to induction by 3-methylcholanthrene and phenobarbitone; the distribution of lactate dehydrogenase, glucose-6-phosphatase, pyruvate kinase and tyrosine aminotransferase activities in the sub-populations is also reported. A marked asymmetry of distribution in all these activities was observed. On the basis of putative zone derivations (based on data of fluorescent dye distribution) of eight factors studied, the distributions of six were consistent with the sub-populations being derived from different acinar zones. Two major discrepancies were noted however, the distribution of pyruvate kinase activity and the response of the sub-populations to phenobarbitone. We conclude from this study that while a metabolic heterogeneity was revealed in the sub-populations generated, further characterisation is required to determine whether acinar zone separation has occurred and if so to what extent.