초록

Apparatus for rapid, ultra sensitive and automatic counting of flourescent biological cells such as microorganisms, carried by a solid support such as a filter.

대표청구항 ▼

[ We claim:] [1.] Apparatus for detecting and counting separated flourescent microorganisms arrayed in two dimensions on a solid support and spaced apart by a distance greater than a diameter of an incident laser beam, said apparatus comprising:a laser light source emitting said incident light beam

[ We claim:] [1.] Apparatus for detecting and counting separated flourescent microorganisms arrayed in two dimensions on a solid support and spaced apart by a distance greater than a diameter of an incident laser beam, said apparatus comprising:a laser light source emitting said incident light beam including means for focusing said light beam into a laser spot having said diameter on the solid support and wherein said laser spot diameter is greater than a size of the microorganism to be detected and wherein said laser spot diameter is between 4 and 14 microns;scanning means for directing said laser spot onto said solid support to irradiate the microorganisms to produce fluorescent spots, wherein a distance between two consecutive scanning lines is such that the entirety of the support is scanned at least twice, by partial overlapping of adjacent scanning paths; andmeans for detecting a resultant fluorescent light at least at one wavelength including means for selecting as samples only detected signals exceeding a given threshold, wherein a set of adjacent samples on a scan line represents a feature;means for providing line-to-line correlation of individual features and establishing a set of correlated-features by comparing features on each pair of adjacent lines in time-synchrony;means for counting a number of lines over which said set of correlated-features occur, each set of correlated features forming an event, and means for eliminating any single uncorrelated-feature;means for comparing said correlated-features on each pair of adjacent lines in time synchrony, at least at two different wavelengths .lambda..sub.1 and .lambda..sub.2 for selecting the correlated-features having an emission intensity ratio at said two wavelengths lower than a predetermined number, said predetermined number being determined by a spectral nature of fluorescent staining and said predetermined number being specified whereby if the emission ratio at said wavelength generated by any correlated samples is greater than a predefined value, said complete event is eliminated;means for determining if, for retained events, the events energy profile in three dimensions is within a predetermined Gaussin shape criteria predetermined by the Gaussian energy profile of the laser beam and rejecting events not within said predetermined Gaussian shaped criteria.