Time-resolved pulsed fluorescence immunometric assays of carcinoembryonic antigen.

A time-resolved pulsed fluorescence immunometric assay (TR-PFIA) for carcinoembryonic antigen is described in which either Eu(III) or Tb(III) chelate is used as label. Described in detail is the assay involving the well-documented format of microtiter matrix and Eu(III) fluorescence enhanced with a beta-diketone and quantified in a commercial time-resolving fluorometer. We have also used the same basic assay, but one with a Tb(III) chelate as label, and we read the fluorescence signal directly off a surface without the application of enhancement solution. The Tb(III) fluorescence is then brought into solution by using an analog of dipicolinic acid in an enhancement solution. The latter approach demonstrates the scope of the methodology, which invokes the extra complexity of enhancement only when increased sensitivity might be required. The power and versatility of the enhancement methodology are demonstrated.

The fluorometric determination of europium ion concentration as used in time-resolved fluoroimmunoassay.

A model of the relationships between optimal fluorescence yield, Eu3+ ion concentration, and the concentration of beta-diketone in the determination of Eu3+ ion concentration in aqueous solutions as used in time-resolved fluoroimmunoassays was developed. The model, in addition to optimizing the determination of the metal ion, also presents a new simple and rapid method for the determination of stability constants for the formation of beta-diketone-Eu3+ complexes. It is shown how the method may be extended to determine the formation constants for Eu3+ with other chelates.

Europium as a label in time-resolved immunofluorometric assays.

A nonisotopic immunoassay has been developed based on a sensitive detection of europium (III) in water solution using time-resolved fluorometry. The europium label is bound to the antibody with EDTA derivatives, either diazophenyl-EDTA-Eu or isothiocyanatophenyl-EDTA-Eu. After the immunometric assay has been completed the europium is preferably dissociated from the antibody at low pH and measured by time-resolved fluorescence in a micellar solution containing Triton X-100, beta-diketone, and a Lewis base. The detergent solubilizes the chelating compounds in the solution and excludes water from the fluorescent ligand-europium complex. Europium concentrations as low as 5 X 10(-14)M were measured using a 1-s counting time. The sensitivity of the immunoassay of rabbit IgG used as a model system was 25 pg/ml (6 pg/assay).

Total body water, total exchangeable sodium and related variables in the Ghanaian.

1. Standard radioisotope dilution techniques employing [3H]water and [22Na]sodium chloride have been used to determine the total body water and total exchangeable sodium of 20 male and 10 female normal Ghanaians (Africans) aged 19--25 years. 2. Lean body mass and total body fat are calculated as a percentage of body weight; the total exchangeable sodium values have been expressed in relation to lean body mass. 3. Comparison of the data for Ghanaian subjects with published figures for Caucasian subjects of similar age shows that the Ghanaian men have much less total body fat and the women a little less total body fat than their Caucasian counterparts. 4. Total exchangeable sodium expressed in terms of lean body mass shows close agreement in both men and women.

Cell inactivation by ultrasound.

Cell survival curves have been obtained for Escherichia coli B (E. coli B) after the sonication of suspensions of the bacteria with continuous wave ultrasound at a fixed frequency of 2 MHZ between peak intensities of 8.7 and 2.25 W cm-2. It was found that under suitable conditions the survival curves were reproducible and it also was found that there was a clear relationship between the rate of inactivation and the peak acoustic intensity of the ultrasound. There appeared to be a lower threshold of peak intensity below which no inactivation was observed.