Method and apparatus for sampling macromolecules from a biological specimen
AssigneeThe Board of Regents of the University of Oklahoma
InventorsJan Sunner, Iwona Beech, Matthew Kowalski, Kathleen Duncan, Joseph Suflita
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Abstract
Apparatus and methods for laser ablation sampling, electrophoretic extraction from the laser plume, electrophoretic transport to a container, and capture of macromolecules of interest. In certain embodiments, when macromolecules of interest are nucleic acid, the apparatus and methods further provides for nucleic acid amplification and detection in a rapid mobile platform for environmental and clinical identification of pathogens.
Claims20
A method of sampling macromolecules of interest directly from a biological specimen for capture and identification, comprising:
- (a) applying a laser beam of photons to the biological specimen to produce an ablation plume comprising a plume matrix and the macromolecules of interest at least partially isolated from the biological specimen;
- (b) extracting the macromolecules of interest from the plume matrix with an electric field;
- and
- (c) capturing the extracted macromolecules of interest in a container for analysis, wherein electric field lines extend from the plume to the container, and wherein an electric potential at the container is higher than the electric potential in the ablation plume, such that negatively charged macromolecules are preferentially extracted from the plume matrix as the macromolecules of interest;
- and wherein the electric field is established by a substrate electrode, a drift electrode and a focusing electrode, and wherein extracting the macromolecules of interest is controlled by increasing the potential of electric fields among the electrodes toward the container.
A method of extracting nucleic acids of interest from a biological specimen, said method comprising:
- (a) ablating the biological specimen with a pulsed laser beam to produce an ablation plume comprising the nucleic acids together with associated plume matrix;
- (b) directing the ablation plume with a first electrode towards a second electrode;
- (c) extracting the nucleic acids from the ablation plume with the second electrode to form transport lines of extracted nucleic acids;
- (d) focusing the transport lines of extracted nucleic acids through an orifice in the second electrode with a third electrode;
- and
- (e) capturing the extracted nucleic acids in a container suitable for conducting a polymerase chain reaction.
An apparatus for extracting macromolecules from a biological specimen, said apparatus comprising:
- (a) a laser configured to ablate the biological specimen with a pulsed laser beam to produce an ablation plume comprising the macromolecules;
- and
- (b) a first electrode, a second electrode, and a third electrode configured to extract the macromolecules from the ablation plume,
- wherein the apparatus is configured to perform a method of sampling the macromolecules of interest directly from the biological specimen for capture and identification, comprising:
- (i) applying the laser beam of photons to the biological specimen to produce the ablation plume further comprising a plume matrix and the macromolecules of interest at least partially isolated from the biological specimen;
- (ii) extracting the macromolecules of interest from the plume matrix with an electric field;
- and
- (iii) capturing the extracted macromolecules of interest in a container for analysis, wherein electric field lines extend from the plume to the container, and wherein the electric potential at the container is higher than the electric potential in the ablation plume, such that negatively charged macromolecules are preferentially extracted from the plume matrix as the macromolecules of interest;
- and wherein the first electrode, the second electrode, and the third electrode, are a substrate electrode, a drift electrode and a focusing electrode, respectively, establishing the electric field, wherein extracting the macromolecules of interest is controlled by increasing the potential of electric fields among the electrodes toward the container.
Dependencies are inferred from the claim text.