Method for rapid detection and evaluation of cultured cell growth
AssigneeOxygen Enterprises, Ltd.
InventorDavid Wilson
Life of this patent
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Abstract
Provided is a method and system for the rapid and accurate detection of growth and metabolism of a cellular microorganism in a population of microorganisms in a non-liquid, culture medium. Further provided is a gelled culture medium containing a non-toxic, water-soluble, phosphorescent compound which measures oxygen content (partial pressure) of an microorganism also contained therein, by oxygen-dependent quenching of phosphorescence; or the gel contains a fluorescent pH indicator that demonstrates growth of the microorganism by pH-dependent intensity change or wavelength shift in the emission spectrum. Further provided is a system and method for killing undesirable microorganisms or colonies in the culture medium without harming the surrounding microorganisms.
Claims26
A method for rapidly detecting growth or metabolism of an immobilized cellular microorganism comprising:
- immobilizing a population of cells of at least one microorganism in an aqueous gelling medium comprising a dissolved oxygen-quenchable phosphorescent compound that is non-toxic to the microorganism;
- thereby
- maintaining communicable contact between each cell and the oxygen-sensitive phosphor in the surrounding medium, such that the phosphorescent compound is responsive to oxygen pressure in the medium;
- exciting said dissolved phosphorescent compound to phosphoresce;
- and
- detecting the phosphorescence in the gelling medium, wherein an increase in phosphorescence is indicative of growth or metabolism of the microorganism.
A method for rapidly detecting growth or metabolism of an immobilized anaerobic microorganism comprising:
- immobilizing a population of cells of at least one anaerobic microorganism in an aqueous gelling medium comprising a dissolved oxygen-quenchable fluorescent pH indicator compound that is non-toxic to the microorganism;
- thereby maintaining communicable contact between each cell and the oxygen-quenchable fluorescent pH indicator in the surrounding medium, such that the quenchable fluorescent pH indicator is responsive to hydrogen ion changes in the medium;
- exciting the dissolved fluorescent indicator to fluoresce, and
- detecting a change in fluorescence emission by the fluorescent pH indicator in the gelled culture medium, wherein a change in fluorescence emission is indicative of a hydrogen ion change resulting from growth or metabolism of the microorganism.
A hollow-form growth chamber for use in rapidly detecting growth or metabolism of a microorganism, comprising:
- a hollow form defining a space;
- and
- an aqueous gelled culture medium within said space of said hollow form,
- wherein said gelled culture medium further comprises a population of cells of at least one microorganism and either a dissolved oxygen-quenchable phosphorescent compound or a dissolved fluorescent pH indicator compound.
A system for killing one or more undesirable microorganism(s) immobilized in a gel within a growth chamber, wherein the system comprises:
- an excitation light source,
- a light detector,
- a collimated laser diode,
- a means for positioning a growth chamber device relative to said collimated laser diode, and
- a processing means operably interconnecting the excitation light source, the light detector and the collimated laser diode.
A kit for use in rapidly detecting growth or metabolism of at least one immobilized microorganism comprising:
- an aqueous gelling medium in powdered form that gels after solublizing;
- a non-toxic, aqueously soluble, oxygen-quenchable phosphorescent compound or fluorescent pH indicator, or both;
- a growth chamber;
- and
- instructional material.
Dependencies are inferred from the claim text.