US 5,018,866 A · Method and apparatus for performing high sensitivity fluorescence measurements

Abstract

An improved method and apparatus is provided for performing high sensitivity fluorescence measurements. A liquid sample contained within a cuvette is exposed to exciting radiation in such a way that the radiation is introduced directly to the sample and is restricted from reaching the container walls. The sample container is in the form of a cylindrical cuvette provided with reflector means on the bottom so that any unabsorbed exciting radiation reaching the cuvette bottom is reflected back through the sample solution. Fluorescent radiation emitted by the sample is collected by an ellipsoidal reflector which substantially surrounds the sample container and focuses radiation incident thereupon onto a small area from which it is efficiently sensed by a photodetetor. The optical arrangement realizes significantly improved signal-to-noise and signal-to-background ratios.

Claims15

Dependent claims

A method of performing fluorescence measurements on a sample capable of being activated by exciting radiation, said method comprising the steps of:

enclosing the sample within a container having
surrounding walls and an open side facing a bottom surface, said bottom surface having reflector means for reflecting any radiation incident thereupon;
introducing a beam of exciting radiation through said open side of container directly onto the sample contained therein in a direction normal to said reflector surface in such a way that said beam does not reach said container walls, thereby avoiding any secondary fluorescence therefrom, any portion of said beam which is unabsorbed by the sample and traverses through the sample to strike said reflector means on si bottom surface being reflected therefrom back through the sample without reaching said container walls, thereby avoiding any secondary fluorescence therefrom;
collecting fluorescent radiation from the sample through collector means surrounding said container walls, said collector means being adapted to focus said fluorescent radiation incident thereupon onto a predefined area;
and
detecting and measuring said fluorescent radiation focused onto said predefined area.

3 more elements

Apparatus for performing fluorescence measurements on a sample by detecting fluorescence induced in the sample by focusing exciting radiation thereupon, said apparatus comprising:

a sample container having surrounding walls and an open side facing a bottom surface, said walls and bottom surface defining a sample-containing cavity, said bottom surface being provided with reflector means for reflecting any radiation incident thereupon;
means for introducing a beam of exciting radiation through said open side of container directly onto the sample contained therein in a direction normal to said reflector surface in such a way that said beam does not reach said container walls, thereby avoiding any secondary fluorescence therefrom, any portion of said beam which is unabsorbed by the sample and traverses through the sample to strike said reflector means on said bottom surface being reflected therefrom back through the sample without reaching said container walls, thereby avoiding any secondary fluorescence therefrom;
collector means for collecting fluorescent radiation emitted from the sample, said collector means substantially surrounding said container walls and being adapted to focus said fluorescent radiation incident thereupon onto a predefined area;
and
means for detecting said focused fluorescent radiation.

2 more elements

Dependencies are inferred from the claim text.

Method and apparatus for performing high sensitivity fluorescence measurements

Life of this patent

Method and apparatus for performing high sensitivity fluorescence measurements

Life of this patent

Abstract

Claims15