High spatial resolution matrix assisted laser desorption/ionization (MALDI)
First Claim
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1. A system for matrix assisted laser desorption/ionization (MALDI) comprising:
- a. a coherent light source to generate light;
b. at least one confocal microscopic objective to create a desorption/ionization source at the surface of a MALDI sample plate adapted to receive a biological sample substrate within the focal working distance of the microscopic objective;
c. at least one fiber optic cable to transport the light to said at least one confocal microscopic objective;
d. at least one collimating fiber optic coupler to collimate the light to an aperture of said at least one fiber optic cable;
e. an insulating microscopic objective holder to hold said at least one confocal microscopic objective and insulate said at least one confocal microscopic objective from the electrical fields of the MALDI;
f. at least one adapter to secure the said objective holder;
g. at least one X, Y positioner to move said confocal microscopic objective in X and Y co-ordinates;
h. a Z positioner to move said confocal microscopic objective in the Z co-ordinate; and
i. a mass analyzer to analyze said sample substrate.
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Abstract
Disclosed is an invention that provides a system and process for focusing light to micron and submicron spot sizes for matrix assisted laser desorption/ionization (MALDI). Moreover, the present invention features a second process and system for creating a correlated optical image of the ion desorption region of a sample substrate.
28 Citations
35 Claims
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1. A system for matrix assisted laser desorption/ionization (MALDI) comprising:
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a. a coherent light source to generate light;
b. at least one confocal microscopic objective to create a desorption/ionization source at the surface of a MALDI sample plate adapted to receive a biological sample substrate within the focal working distance of the microscopic objective;
c. at least one fiber optic cable to transport the light to said at least one confocal microscopic objective;
d. at least one collimating fiber optic coupler to collimate the light to an aperture of said at least one fiber optic cable;
e. an insulating microscopic objective holder to hold said at least one confocal microscopic objective and insulate said at least one confocal microscopic objective from the electrical fields of the MALDI;
f. at least one adapter to secure the said objective holder;
g. at least one X, Y positioner to move said confocal microscopic objective in X and Y co-ordinates;
h. a Z positioner to move said confocal microscopic objective in the Z co-ordinate; and
i. a mass analyzer to analyze said sample substrate. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A system for focusing light to a less than 0.5 micron spot size area for matrix assisted laser desorption/ionization (MALDI) comprising:
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a. a coherent light source to generate ultra-violet light;
b. at least one confocal microscopic objective to create a desorption/ionization source of less than 0.5 micron spatial resolution at the surface of a MALDI sample plate adapted to receive a biological sample substrate within the focal working distance of the microscopic objective;
c. at least one fiber optic cable to transport the ultra-violet light to said at least one confocal microscopic objective;
d. at least one collimating fiber optic coupler to collimate the light to the aperture of said at least one fiber optic cable;
e. at least one insulating microscopic objective holder to hold said at least one confocal microscopic objective and insulate said at least one confocal microscopic objective from electrical fields of the MALDI;
f. at least one adapter to secure the said objective holder;
g. at least one X, Y positioner to move said confocal microscopic objective in X and Y coordinates;
h. a Z positioner to move said confocal microscopic objective in the Z co-ordinate; and
i. a mass analyzer to analyze ions desorbed from said sample substrate. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18)
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19. A process for focusing a light source to a sub-micron spot size for matrix assisted laser desorption/ionization (MALDI), comprising the steps of:
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a. depositing a biological sample substrate containing analyte and an appropriately absorbing matrix on a sample plate;
b. generating a coherent light source;
c. positioning said sample plate within the focal working distance of at least one confocal microscopic objective;
d. coupling said at least one confocal microscopic objective to said coherent light source with at least one fiber optic cable;
e. positioning said at least one confocal microscopic objective in a geometry that does not interfere with the path of desorbed sample ions;
f. focusing said coherent light source through said at least one microscopic objective to create a desorption/ionization ultra-violet source of submicron spatial resolution directed at said sample substrate;
g. ionizing said sample substrate; and
h. separating and detecting ions from said ionized sample substrate in one or more stages using an appropriate mass separation and analysis method. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27)
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28. A process for creating a correlated optical image of the ion desorption region associated with matrix assisted laser desorption/ionization (MALDI) of a biological sample substrate comprising the steps of:
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a. depositing a biological sample substrate containing analyte and an appropriately absorbing matrix on a sample plate;
b. generating a coherent light source;
c. positioning said sample plate within the focal working distance of at least one confocal microscopic objective;
d. coupling said at least one confocal microscopic objective to said coherent light source with at least one fiber optic cable;
e. positioning said at least one confocal microscopic objective in a geometry that does not interfere with the path of desorbed sample ions;
f. focusing said coherent light source through said at least one microscopic objective to create a desorption/ionization ultra-violet light source of submicron spatial resolution directed at said sample substrate;
g. ionizing said sample substrate; and
h. separating and detecting ions from said ionized sample substrate in one or more stages using an appropriate mass separation and analysis method;
i. illuminating the sample;
j. transferring an optical image of the ionized sample substrate using said at least one fiber optic cable; and
k. capturing an optical image of said ionized sample substrate. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35)
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Specification