Experimental determination of the linear correlation between in vivo TV fluorescence intensity and vascular and tissue FITC-Dx concentrations

Document Type

Article

Publication Date

1-1-1991

Abstract

A novel in vivo calibration procedure was developed to determine the microvascular and tissue concentration of fluorescein isothiocyanate-labeled dextrans (FITC-Dx) in the hamster cheek pouch, using intravital fluorescence microscopy with manually controlled TV camera gain and threshold value. Two FITC-dextrans (70,000 and 150,000 MW) were used as tracers. Five minutes after the tracer was administered, selected venules (diameter 20-50 μm) were videotaped, and intravascular gray levels were obtained by digital image processing. Simultaneously, arterial blood samples were taken to measure vascular FITC-Dx concentrations with a spectrofluorometer. The gray levels and the concentrations were used to produce a calibration curve for the vascular FITC-Dx concentration. A similar calibration curve for the interstitial FITC-Dx concentration was obtained by first video recording interstitial space areas saturated with the tracer. After flushing out the tracer in the vessels, the hamster cheek pouch was then cut, weighed, and homogenized. The interstitial FITC-Dx concentration was finally measured with a spectrofluorometer. The gray levels and the concentrations were used to produce a calibration curve for the interstitial FITC-Dx concentration. The gray level was found to vary linearly with both the FITC-Dx vascular concentration (range 0.4-3.0 mg/ml) and the interstitial FITC-Dx concentration (0.12-1.50 mg/ml) in the hamster cheek pouch. © 1991.

Identifier

0026008608 (Scopus)

Publication Title

Microvascular Research

External Full Text Location

https://doi.org/10.1016/0026-2862(91)90087-R

e-ISSN

10959319

ISSN

00262862

PubMed ID

1719355

First Page

198

Last Page

208

Issue

2

Volume

42

Grant

R01HL043146

Fund Ref

National Heart, Lung, and Blood Institute

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