US20030072684A1 - Devices for use in MALDI mass spectrometry - Google Patents

Devices for use in MALDI mass spectrometry Download PDF

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Publication number
US20030072684A1
US20030072684A1 US10/302,984 US30298402A US2003072684A1 US 20030072684 A1 US20030072684 A1 US 20030072684A1 US 30298402 A US30298402 A US 30298402A US 2003072684 A1 US2003072684 A1 US 2003072684A1
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United States
Prior art keywords
plate
alignment
sample
samples
sample plate
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Abandoned
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US10/302,984
Inventor
N. Anderson
John Lennon
Jack Goodman
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Thermo Finnigan LLC
Original Assignee
Anderson N. Leigh
Lennon John Joseph
Jack Goodman
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Application filed by Anderson N. Leigh, Lennon John Joseph, Jack Goodman filed Critical Anderson N. Leigh
Priority to US10/302,984 priority Critical patent/US20030072684A1/en
Publication of US20030072684A1 publication Critical patent/US20030072684A1/en
Assigned to THERMO FINNIGAN LLC reassignment THERMO FINNIGAN LLC ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: LARGE SCALE PROTEOMICS CORPORATION
Abandoned legal-status Critical Current

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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00Supporting devices; Holding devices
    • B01L9/54Supports specially adapted for pipettes and burettes
    • HELECTRICITY
    • H01ELECTRIC ELEMENTS
    • H01JELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
    • H01J49/00Particle spectrometers or separator tubes
    • H01J49/0027Methods for using particle spectrometers
    • H01J49/0031Step by step routines describing the use of the apparatus
    • HELECTRICITY
    • H01ELECTRIC ELEMENTS
    • H01JELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
    • H01J49/00Particle spectrometers or separator tubes
    • H01J49/02Details
    • H01J49/04Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components
    • H01J49/0409Sample holders or containers
    • H01J49/0418Sample holders or containers for laser desorption, e.g. matrix-assisted laser desorption/ionisation [MALDI] plates or surface enhanced laser desorption/ionisation [SELDI] plates
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02Adapting objects or devices to another
    • B01L2200/025Align devices or objects to ensure defined positions relative to each other
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/02Devices for withdrawing samples
    • G01N1/22Devices for withdrawing samples in the gaseous state
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/04Details of the conveyor system
    • G01N2035/0474Details of actuating means for conveyors or pipettes
    • G01N2035/0491Position sensing, encoding; closed-loop control
    • G01N2035/0494Detecting or compensating piositioning errors
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027General features of the devices
    • G01N2035/1034Transferring microquantities of liquid
    • G01N2035/1039Micropipettes, e.g. microcapillary tubes
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/028Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations having reaction cells in the form of microtitration plates
    • HELECTRICITY
    • H01ELECTRIC ELEMENTS
    • H01JELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
    • H01J49/00Particle spectrometers or separator tubes
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00Chemistry: analytical and immunological testing
    • Y10T436/25Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/2575Volumetric liquid transfer

Definitions

  • the invention relates to a method and apparatus for preparation of samples that are to be subjected to mass spectrometry analysis. Specifically, the invention relates to methods for preparing samples for mass spectrometry analysis and to plate-like members used to position samples on a sample plate for subsequent mass spectrometry analysis.
  • Mass spectrometry devices measure the molecular mass of a molecule by measuring the molecule's flight path through a set of magnetic and electric fields. Such devices are well known and are widely used in the field of bio-molecular research. In proteomics research, for instance, mass spectrometry is used to identify proteins.
  • Proteins are typically separated from one another by electrophoresis, such as the techniques described and claimed U.S. Pat. No. 5,993,627 to Anderson et. al. (the Anderson et. al. patent), which is incorporated herein by reference in its entirety.
  • a tissue sample is first subjected to a first dimension electrophoresis process where groups of proteins are separated linearly within a tubular gel filled column.
  • the first dimension separation of proteins is then inserted along an edge of a flat planar gel slab and subjected to a second dimension of electrophoresis thereby generating a two dimensional pattern of spots formed by clusters of proteins that have moved to respective iso-electric focusing points.
  • selected proteins are excised from the second dimension gel slab for further study.
  • the selected excised spots are next prepared for analysis using, for instance, mass spectrometry.
  • An increasingly used technique for studying biological molecules includes the use of MALDI mass spectrometry apparatus (matrix-assisted laser desorption ionization apparatus) where the biological sample is embedded in a volatile matrix and is vaporized by being subjected to an intense laser emission.
  • MALDI apparatus matrix-assisted laser desorption ionization apparatus
  • MALDI-TOF apparatus TOF is time-of-flight spectrometry.
  • MALDI-TOF techniques are used to determine the molecular weight of peptides produced by digestion of isolated proteins.
  • MALDI-TOF apparatus is VOYAGER DE STR Biospectrometry Workstation manufactured and sold by APPLIED BIOSYSTEMS.
  • FIG. 1 depicts a generic MALDI-TOF apparatus that includes a frame 1 that supports the electronic and computer equipment necessary to control a laser 5 .
  • the laser 5 is aimed at a fixed location in a positioning mechanism 10 .
  • the positioning mechanism 10 includes means (not shown) for positioning a sample in the line of fire of the laser 5 .
  • the laser is fixed in place and the sample is moved into position for analysis.
  • the depicted MALDI-TOF apparatus includes a small removable sample plate 15 , shown in FIG. 2, that fits into the positioning mechanism 10 .
  • the sample plate 15 is insertable into a slot 20 in the positioning mechanism 10 of the MALDI-TOF apparatus and is thereafter held in a specific orientation within the positioning mechanism 10 for sample analysis.
  • the sample plate 15 typically holds a plurality of discrete samples on one surface thereof, with the samples being spaced apart from one another.
  • the sample plate 15 includes guide members 15 a , guide holes 15 b and alignment pin 15 d that are used by corresponding members (not shown) within the positioning mechanism 10 for positioning and moving the sample plate 15 with respect to the line of fire of the laser 5 .
  • the samples are typically loaded onto the sample plate 15 by a separate device or robotic apparatus, such as the X-Y robotic apparatus depicted in FIG. 3.
  • a separate device or robotic apparatus such as the X-Y robotic apparatus depicted in FIG. 3.
  • Such X-Y robotic apparatuses are typically manufactured and sold with each specific mass spectrometry apparatus.
  • the X-Y robotic apparatus depicted in FIG. 3 includes a recess that retains the sample plate 15 in position for sample loading.
  • the X-Y robotic apparatus includes a first arm 30 that moves back and forth along an X axis and a second arm 40 that moves along a Y axis defined along the length of the first arm.
  • the second arm 40 supports a pipette tip 45 that is used to spot samples on the sample plate 15 .
  • the pipette tip 45 is moved by the first and second arms 30 and 40 to a position above a 96-well plate 50 (or other similar sample holder) or microcentrifuge tubes and the pipette tip 45 picks up a sample from the 96-well plate 50 or microcentrifuge tubes. The pipette tip 45 is then moved to a location above the sample plate 15 and the sample is spotted on the specified location of the sample plate.
  • an array of samples are spotted on the sample plate 15 at predetermined locations. After the array of samples are loaded onto the sample plate 15 , the sample plate 15 is inserted into the slot 20 of the MALDI apparatus. Using an imaging system 25 focused on the sample plate 15 within the MALDI apparatus, in combination with the positioning mechanism 10 , the laser beam from the laser 5 can be aimed, one by one, at the sample(s) on the sample plate 15 .
  • the mass spectrometry apparatus typically takes several hours to analyze an array of samples on the sample plate 15 . Therefore, in order to minimize human involvement, automation of the process is critical.
  • the locations of the samples are typically pre-programmed into the computer that controls the MALDI-TOF apparatus so that during the analysis of the samples, the positioning mechanism 10 automatically repositions the sample plate 15 into the line of fire of the laser 5 . Therefore, if any of the samples on the sample plate 15 are not properly positioned, the laser 5 is not likely to hit each of the samples.
  • a 10 ⁇ 10 array of samples is positioned on the upper surface at spaced apart intervals.
  • the positioning mechanism 10 moved into a target position with respect to centers of the desired location of each spot. The desired location of each spot assumes that center of each of the spots in the 10 ⁇ 10 array is constant.
  • the positioning mechanism 10 within the MALDI apparatus has positional accuracy with respect to movement of the sample plate 15
  • the X-Y robotic apparatus typically sold with a MALDI apparatus is not as precise with respect to accurate spotting or depositing of samples on the sample plate 15 .
  • the spots in a 10 ⁇ 10 array of samples do not wind up being centered on the desired center targeted by the positioning mechanism 10 .
  • the array of 10 ⁇ 10 samples may have some samples that are accurately centered, and other samples that are off center by as much as half the width of the sample.
  • Another problem relates to the pre-programmed settings for locating samples on sample plates 15 of such X-Y robotic apparatuses.
  • X-Y robotic apparatuses are not programmed to maximize the use of the space of the surface of the sample plate 15 .
  • each pair of adjacent samples in the 10 ⁇ 10 array of samples mentioned above is typically spaced apart by a distance greater than the diameter of the sample.
  • the MALDI-TOF apparatus takes considerable time to analyze samples and requires human interaction to switch sample plates 15 in the mass spectrometer, it is advantageous to analyze as many samples as possible on a single sample plate. Current technology limits the number of samples that can be analyzed on a single sample plate 15 .
  • One object of the present invention is to provide a means for more precisely depositing samples on a sample plate of a mass spectrometry apparatus.
  • Another object of the present invention is to provide a means for maximizing the number of samples spotted on the surface of a sample plate of a mass spectrometry apparatus.
  • An aspect of the present invention relates to the use of an alignment plate to assist in accurate positioning of a pipette tip delivering samples onto a sample plate of a mass spectrometer.
  • Another aspect of the present invention relates to the use of an adaptor plate and an alignment plate with an X-Y manipulator device originally designed for liquid sample manipulation.
  • Use of the adaptor plate and alignment plate of the present invention allows for new usage of the X-Y manipulator device.
  • the X-Y manipulator device can be used for spotting samples on a sample plate of a mass spectrometer with the alignment providing alignment holes for accurate positioning of samples on the sample plate.
  • a plurality of alignment plates are used to guide a pipette tip toward a sample plate for accurate positioning of samples on the sample plate.
  • a first alignment plate guides the pipette tip to spot an array of samples on the sample plate.
  • a second array of samples can be used to guide the pipette tip to spot a second array of samples on the sample plate, where the second array of samples are located between, but offset from the first array of samples thereby maximizing the number of samples positionable on the sample plate.
  • FIG. 1 is a side schematic view of a mass spectrometry apparatus
  • FIG. 2 is a perspective view of a sample plate for samples to be analyzed in the mass spectrometry apparatus depicted in FIG. 1;
  • FIG. 3 is a perspective view of an X-Y robotic apparatus used to spot samples on the surface of the sample plate depicted in FIG. 2;
  • FIG. 4 is top view of an alignment plate having a 10 ⁇ 10 array of alignment holes used for positioning samples on a sample plate in accordance with one embodiment of the present invention
  • FIG. 5 is side exploded cross-section view taken along the line V-V in FIG. 4 showing the sample plate mounted on the table of an X-Y robotic apparatus, with the alignment plate depicted in FIG. 4 being mounted thereon in accordance with the present invention
  • FIG. 6 is a side cross-section view taken along the line VI-VI in FIG. 4 showing the sample plate mounted on the table of an X-Y robotic apparatus, with the alignment plate depicted in FIGS. 4 and 5 mounted thereon, and a pipette delivering a sample through a hole in the alignment plate to the surface of the sample plate in accordance with the present invention;
  • FIG. 7 is a top view of the sample plate with an array of 10 ⁇ 10 samples accurately positioned on the upper surface thereof in accordance with the present invention
  • FIG. 8 is top view of a second alignment plate having a 9 ⁇ 9 array of alignment holes used for positioning samples on a sample plate in accordance with the present invention, with the 9 ⁇ 9 array of alignment holes being located to compliment the 10 ⁇ 10 array of holes in the alignment plate depicted in FIG. 4;
  • FIG. 9 is another top view of the sample plate depicted in FIG. 7 with the array of 10 ⁇ 10 samples accurately positioned on the upper surface thereof along with an additional 9 ⁇ 9 array of samples loaded using the alignment plate depicted in FIG. 8 in accordance with the present invention
  • FIG. 10 is a top view of an adaptor plate for use with various types of X-Y robotic apparatus in accordance with a second embodiment of the present invention.
  • FIG. 11 is a top view of an alignment plate in accordance with the second embodiment of the present invention.
  • FIG. 12 is a side view showing two sample plates mounted on the adaptor plate depicted in FIG. 10, with the alignment plate depicted in FIG. 11 mounted thereon, and a pipette delivering a sample through a hole in the alignment plate to the surface of one of the sample plates in accordance with the present invention
  • FIG. 13 is an exploded perspective view showing the alignment plate, one sample plate and the adaptor plate, where the alignment plate is provided with a 10 ⁇ 10 array of alignment holes in accordance with the present invention
  • FIG. 14 is an exploded perspective view showing another alignment plate, one sample plate and the adaptor plate, where the alignment plate is provided with a 9 ⁇ 9 array of alignment holes in accordance with the present invention.
  • FIG. 15 is a block diagram showing the X-Y robotic apparatus and a computer for controlling the X-Y robotic apparatus in accordance with the present invention.
  • the present invention relates to alignment plates that include holes for guiding a pipette tip toward a sample plate thereby assisting in accurately spotting or depositing samples on a sample plate of a mass spectrometry apparatus.
  • FIG. 4 is top view of an alignment plate 60 that is formed with a plurality of holes 61 that define an array of holes. In the embodiment depicted in FIG. 4, there are one hundred holes 61 defining a 10 ⁇ 10 array of alignment holes used for positioning samples on a sample plate 15 in accordance with one embodiment of the present invention.
  • Each of the holes 61 extends from an upper surface 62 of the alignment plate 61 to a lower surface 65 .
  • each hole 61 is further provided with a conical contour 63 whose function is described in greater detail below.
  • the alignment plate 61 is further formed with a plurality of positioning legs 68 that extend downward away from the upper surface 62 .
  • the X-Y robotic apparatus described above with respect to FIG. 3 is typically formed with means for retaining the sample plate 15 .
  • the X-Y robotic apparatus includes a table 48 that includes a recess 49 for receiving the sample plate 15 , as shown in FIGS. 5 and 6.
  • the recess 49 snuggly receives the sample plate 15 thereby retaining the sample plate 15 in a stationary position.
  • the alignment plate 60 and its positioning legs 68 are formed to conform to the surface contours of the table 48 such that the sample plate 15 may be sandwiched in between the table 48 and the alignment plate 60 , as depicted in FIG. 6.
  • the positioning legs 68 firmly engage the contours of the table 48 such that the alignment plate 60 is held in a stationary position above the sample plate 15 , as depicted in FIG. 6.
  • the positioning legs 68 may be dimensioned to contact the outer edges of the sample plate 15 .
  • FIG. 6 is a side view showing the sample plate mounted on the table of the X-Y robotic apparatus, with the alignment plate 60 mounted thereon.
  • a pipette tip 45 is shown extending through one of the holes 61 in the alignment plate 60 delivering the sample to the upper surface 62 of the sample plate 60 .
  • the X-Y robotic apparatus depicted in FIG. 3 positions the pipette tip 45 , above the sample plate 15 in order to deposit a sample.
  • the pipette tip 45 engages the conical contour 63 of the hole 61 in the alignment plate 60 .
  • FIG. 7 is a top view of the sample plate 15 with an array of 10 ⁇ 10 samples accurately positioned on the upper surface of the sample plate 15 .
  • the pipette tip 45 is guided into and through the holes 61 in the alignment plate 60 in order to more reliably position the samples on the sample plate 15 .
  • the alignment plate 60 depicted in FIGS. 4 and 5 is not limited to the 10 ⁇ 10 array of holes 61 . Alternatively, a greater number of holes may be formed. For instance, the inventors have made an alternate embodiment of the alignment plate that has a 17 ⁇ 17 array of holes (not shown).
  • FIG. 8 is top view of a second alignment plate 70 having a 9 ⁇ 9 array of alignment holes used for positioning samples on the sample plate 15 in accordance with an additional feature of the present invention.
  • the 9 ⁇ 9 array of alignment holes 61 are located to compliment the 10 ⁇ 10 array of holes in the alignment plate 60 depicted in FIG. 4. Specifically, after the 10 ⁇ 10 array of samples is deposited on the sample plate 15 , as shown in FIG. 7, the alignment plate 60 is removed from the table 48 . The second alignment plate 70 is then installed on the table 48 above the sample plate 15 having the 10 ⁇ 10 array of samples (FIG. 7) deposited thereon.
  • An additional eighty-one (81) samples are then deposited on to the sample plate 15 to form the two space apart arrays of samples on the sample plate 15 , as shown in FIG. 9.
  • the alignment plate 60 is used to accurately position the pipette tip 45 above the sample plate 15 in order to deposit a 10 ⁇ 10 array of samples on the sample plate 15 .
  • the second alignment plate 60 is used to accurately position the pipette tip 45 above the sample plate 15 in order to deposit an additional 9 ⁇ 9 array of samples between the 10 ⁇ 10 array of samples, as depicted in FIG. 9.
  • the second alignment plate 70 can have any number of holes in order to complement the holes in the first alignment plate 60 .
  • the second alignment plate 70 is provided with either a 11 ⁇ 11 array of alignment holes or a 13 ⁇ 13 array of alignment holes to significantly increase the number of samples deposited on the sample plate 15 .
  • the second alignment plate 70 described above and shown in the drawings includes an array of holes that are configured to be diagonally offset from the holes in the first alignment plate.
  • the array of holes in the second alignment plate may be offset from the holes in the first plate along one axis, not diagonally offset.
  • a series of alignment plates can be configured to assist in spotting three or four separate arrays of spots on a sample plate, with the alignment holes in each alignment plate being offset from the other alignment plate holes.
  • the sample plate 15 is loaded with samples by an X-Y robotic apparatus that is not provided with a recess for retaining the sample holder 15 .
  • an adaptor plate 80 depicted in FIG. 10 is used to retain the sample holder 15 .
  • the adaptor plate 80 has an outer perimeter 82 that is formed with dimensions corresponding to retainer portions 85 (shown in FIGS. 13 and 14) on an X-Y robotic apparatus.
  • the adaptor plate 80 is provided in order to deposit samples on the sample plate 15 using an X-Y robotic apparatus that is not originally manufactured for use with a MALDI-TOF apparatus.
  • FIG. 10 is a top view of the adaptor plate 80 .
  • the adaptor plate 80 is formed with at least one recess 49 ′ that is shaped to securely receive and retain the sample plate 15 .
  • the recess 49 ′ and the recess 49 described above have generally the same shape.
  • the recess 49 ′ (and the recess 49 in FIGS. 5 and 6) includes an alignment recess 49 d that receives an alignment pin 15 d such that the sample plate 15 only fits into the recess 49 ′ in a single orientation.
  • the adaptor plate 80 is formed with two recesses 49 ′ but could be formed with more than two recesses 49 ′ providing the adaptor plate 80 is large enough to receive such recesses.
  • the size of the adaptor plate 80 is dependent upon the size of the retainer portions 85 of the X-Y robotic apparatus, since the adaptor plate 80 is made to be received in the retainer portions 85 .
  • a first alignment plate 90 shown in FIG. 11, is dimensioned to correspond to the size and shape of the adaptor plate 80 . Since the adaptor plate 80 is formed with two recesses 49 ′ to receive and securely retain two sample plates 15 , the first alignment plate 90 is formed with two separate 10 ⁇ 10 arrays of alignment holes 91 . Each two 10 ⁇ 10 arrays of alignment holes 91 corresponds to one of the sample plates 15 . Each alignment hole 91 is formed with a conical contour 92 to guide a pipette as the pipette deposits a sample on the sample plate 15 . The first alignment plate 90 is further formed with positioning legs 94 for holding the first alignment plate 90 on the adaptor plate 80 .
  • FIG. 12 is a slightly exploded side view showing two sample plates 15 mounted on the adaptor plate 80 , with the first alignment plate 90 depicted in FIG. 11 mounted thereon. A pipette tip 45 is shown extending through one of the alignment holes 91 depositing a sample on surface of one of the sample plates 15 .
  • FIG. 13 is an exploded perspective view showing the first alignment plate 90 , one sample plate 15 and the adaptor plate 80 , where the first alignment plate is provided with a 10 ⁇ 10 array of alignment holes.
  • a second alignment plate 98 shown in FIG. 14 is provided with a 9 ⁇ 9 array of alignment holes in order to load a second array of samples onto the sample plate 15 .
  • first and second alignment plates 90 and 98 allow multiple arrays of samples to be deposited on a single sample plate 15 . Further the adaptor plate 80 , in combination with the first and second alignment plates 90 and 98 allows for two sample plates 15 to be loaded with multiple arrays of samples.
  • alignment holes in the alignment plates of the present invention is not limited to the specific number shown in the drawings.
  • the alignment plates of the present invention may be provided with an array of holes of over to 20 ⁇ 20 holes if desired and the size of the sample plate allows. The number of holes in any array of holes is dependent primarily upon the size of the sample plate and the size of the samples to be deposited on the sample plate.
  • the present invention is not limited to the specific sample plate depicted.
  • the present invention is applicable to all.
  • the sample plate depicted is about 2.25 inches by 2.25 inches in size.
  • Other MALDI apparatus' use sample plates that are larger and can hold more samples.
  • the present invention is applicable to larger sample plates as well.
  • the alignment plates described above can be dimensioned to fit any sample plate.
  • samples to be deposited on the sample plate 15 can be prepared in a variety of ways.
  • the samples can be based upon processed spots initially excised from a 2-D electrophoresis gel slab.
  • 2-D electrophoresis gel slabs are prepared in a variety of ways, for instance see U.S. Pat. No. 5,993,627, which is incorporated herein by reference in its entirety.
  • the spot is deposited in a 96-well microtiter plate (aka 96-well plate 50 depicted in FIG. 3) or microcentrifuge tube.
  • the spot may include a specific peptide or protein that is to be subjected to mass spectrometry analysis.
  • the 96-well plate 50 or microcentrifuge tube is then moved to an X-Y robotic apparatus, such as the X-Y robotic apparatus 48 described above with respect to FIG. 3.
  • the X-Y robotic apparatus 48 is typically controlled by a computer 100 , as shown in FIG. 15.
  • the computer 100 typically includes an input 105 such as a keyboard, floppy disk drive and or mouse.
  • the computer 100 also includes a display 110 and an output 115 .
  • the X-Y robotic apparatus 48 under the control of the computer 100 , performs several procedures on each of the spots in the 96-well plate 50 in order to prepare the excised spots for spotting on the sample plate.
  • the spot which includes gel slab material, must be digested in order to remove the unwanted gel material.
  • Such procedures are well known in the art and vary depending upon various circumstances. However, almost all digestion procedures include manipulation of fluid in and about the 96-well plate 50 retaining the samples.
  • each sample is typically moved to a clean 96-well plate by the X-Y robotic device 48 .
  • the computer 100 records identification of the sample and its location on the sample plate 15 in memory (not shown) and is configured to output location information upon request.
  • spots excised from a 2-D gel are processed and thereafter deposited on a sample plate 15 for analysis in the MALDI apparatus 1 .
  • An advantage of the present invention is that the X-Y robotic apparatus 48 that performs the liquid manipulation in the sample preparation procedures may now be used to spot the samples on the sample plate 15 using the alignment plates and adaptor plate described above.

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  • Biochemistry (AREA)
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Abstract

An alignment plate is provided with a plurality of holes for guiding a pipette tip toward a sample plate of a MALDI mass spectrometer. Each of the holes is provided with a conical upper contour in order to guide the pipette tip toward a specific location on the sample plate. Two companion alignment plates are used in order to overlay two separate arrays of samples on the sample plate. For instance, a first of two alignment plates is formed with a 10×10 array of holes so that a 10×10 array of samples is deposited by the pipette tip onto the sample plate. The second of the two alignment plates is formed with a 9×9 array of holes so that a 9×9 array of samples is deposited on the sample plate at locations offset from the 10×10 array of samples already on the sample plate. The number of samples loaded on the sample plate is large and the space on the sample plate is more fully utilized.

Description

  • This application is a continuation of U.S. application Ser. No. 09/644,780, filed Aug. 24, 2000, which claims priority to U.S. Provisional Application No. 60/151,075, filed Aug. 27, 1999; the contents of both applications being incorporated herein by reference.[0001]
  • BACKGROUND OF THE INVENTION
  • 1. Field of the Invention [0002]
  • The invention relates to a method and apparatus for preparation of samples that are to be subjected to mass spectrometry analysis. Specifically, the invention relates to methods for preparing samples for mass spectrometry analysis and to plate-like members used to position samples on a sample plate for subsequent mass spectrometry analysis. [0003]
  • 2. Background Information [0004]
  • Mass spectrometry devices measure the molecular mass of a molecule by measuring the molecule's flight path through a set of magnetic and electric fields. Such devices are well known and are widely used in the field of bio-molecular research. In proteomics research, for instance, mass spectrometry is used to identify proteins. [0005]
  • Proteins are typically separated from one another by electrophoresis, such as the techniques described and claimed U.S. Pat. No. 5,993,627 to Anderson et. al. (the Anderson et. al. patent), which is incorporated herein by reference in its entirety. For instance, as set forth in the Anderson patent, a tissue sample is first subjected to a first dimension electrophoresis process where groups of proteins are separated linearly within a tubular gel filled column. The first dimension separation of proteins is then inserted along an edge of a flat planar gel slab and subjected to a second dimension of electrophoresis thereby generating a two dimensional pattern of spots formed by clusters of proteins that have moved to respective iso-electric focusing points. Thereafter, selected proteins are excised from the second dimension gel slab for further study. The selected excised spots are next prepared for analysis using, for instance, mass spectrometry. [0006]
  • An increasingly used technique for studying biological molecules includes the use of MALDI mass spectrometry apparatus (matrix-assisted laser desorption ionization apparatus) where the biological sample is embedded in a volatile matrix and is vaporized by being subjected to an intense laser emission. One such MALDI apparatus is a MALDI-TOF apparatus (TOF is time-of-flight spectrometry). In the field of proteomics, mass spectrometry, and in particular, MALDI-TOF techniques are used to determine the molecular weight of peptides produced by digestion of isolated proteins. One such MALDI-TOF apparatus is VOYAGER DE STR Biospectrometry Workstation manufactured and sold by APPLIED BIOSYSTEMS. [0007]
  • FIG. 1 depicts a generic MALDI-TOF apparatus that includes a [0008] frame 1 that supports the electronic and computer equipment necessary to control a laser 5. The laser 5 is aimed at a fixed location in a positioning mechanism 10. The positioning mechanism 10 includes means (not shown) for positioning a sample in the line of fire of the laser 5. Typically in a MALDI-TOF apparatus, the laser is fixed in place and the sample is moved into position for analysis. The depicted MALDI-TOF apparatus includes a small removable sample plate 15, shown in FIG. 2, that fits into the positioning mechanism 10. Typically, the sample plate 15 is insertable into a slot 20 in the positioning mechanism 10 of the MALDI-TOF apparatus and is thereafter held in a specific orientation within the positioning mechanism 10 for sample analysis. The sample plate 15 typically holds a plurality of discrete samples on one surface thereof, with the samples being spaced apart from one another. The sample plate 15 includes guide members 15 a, guide holes 15 b and alignment pin 15 d that are used by corresponding members (not shown) within the positioning mechanism 10 for positioning and moving the sample plate 15 with respect to the line of fire of the laser 5.
  • The samples are typically loaded onto the [0009] sample plate 15 by a separate device or robotic apparatus, such as the X-Y robotic apparatus depicted in FIG. 3. Such X-Y robotic apparatuses are typically manufactured and sold with each specific mass spectrometry apparatus. The X-Y robotic apparatus depicted in FIG. 3 includes a recess that retains the sample plate 15 in position for sample loading. The X-Y robotic apparatus includes a first arm 30 that moves back and forth along an X axis and a second arm 40 that moves along a Y axis defined along the length of the first arm. The second arm 40 supports a pipette tip 45 that is used to spot samples on the sample plate 15. Specifically, the pipette tip 45 is moved by the first and second arms 30 and 40 to a position above a 96-well plate 50 (or other similar sample holder) or microcentrifuge tubes and the pipette tip 45 picks up a sample from the 96-well plate 50 or microcentrifuge tubes. The pipette tip 45 is then moved to a location above the sample plate 15 and the sample is spotted on the specified location of the sample plate.
  • Typically, an array of samples are spotted on the [0010] sample plate 15 at predetermined locations. After the array of samples are loaded onto the sample plate 15, the sample plate 15 is inserted into the slot 20 of the MALDI apparatus. Using an imaging system 25 focused on the sample plate 15 within the MALDI apparatus, in combination with the positioning mechanism 10, the laser beam from the laser 5 can be aimed, one by one, at the sample(s) on the sample plate 15.
  • The mass spectrometry apparatus typically takes several hours to analyze an array of samples on the [0011] sample plate 15. Therefore, in order to minimize human involvement, automation of the process is critical. The locations of the samples are typically pre-programmed into the computer that controls the MALDI-TOF apparatus so that during the analysis of the samples, the positioning mechanism 10 automatically repositions the sample plate 15 into the line of fire of the laser 5. Therefore, if any of the samples on the sample plate 15 are not properly positioned, the laser 5 is not likely to hit each of the samples. For instance, on the sample plate 15 depicted in FIG. 2, a 10×10 array of samples is positioned on the upper surface at spaced apart intervals. The positioning mechanism 10 moved into a target position with respect to centers of the desired location of each spot. The desired location of each spot assumes that center of each of the spots in the 10×10 array is constant.
  • Unfortunately, there are several shortcomings associated with the above described X-Y robotic apparatus (FIG. 3). Although the [0012] positioning mechanism 10 within the MALDI apparatus has positional accuracy with respect to movement of the sample plate 15, the X-Y robotic apparatus typically sold with a MALDI apparatus is not as precise with respect to accurate spotting or depositing of samples on the sample plate 15. Specifically, the spots in a 10×10 array of samples do not wind up being centered on the desired center targeted by the positioning mechanism 10. The array of 10×10 samples may have some samples that are accurately centered, and other samples that are off center by as much as half the width of the sample.
  • Part of the problem with such X-Y robotic apparatuses relates to the [0013] replaceable pipette tips 45 used to retrieve a sample and deposit the sample onto the sample plate 15. The pipette tips are not perfectly uniform in size and shape. Further, the tips are deformable and hence accurate positioning of samples is not possible. As well, the X-Y robotic apparatus may not have movement and location capabilities as precise as the movement and location capabilities of the positioning mechanism 10 of the MALDI apparatus.
  • Another problem relates to the pre-programmed settings for locating samples on [0014] sample plates 15 of such X-Y robotic apparatuses. Specifically, X-Y robotic apparatuses are not programmed to maximize the use of the space of the surface of the sample plate 15. For instance, each pair of adjacent samples in the 10×10 array of samples mentioned above is typically spaced apart by a distance greater than the diameter of the sample. There is a substantial amount of empty space on the surface of the sample plate 15 even with a 10×10 array of samples. Since the MALDI-TOF apparatus takes considerable time to analyze samples and requires human interaction to switch sample plates 15 in the mass spectrometer, it is advantageous to analyze as many samples as possible on a single sample plate. Current technology limits the number of samples that can be analyzed on a single sample plate 15.
  • There is a need for more precise positioning of samples on a sample plate and a need for maximizing the space on a sample plate in order to more efficiently utilize a mass spectrometry apparatus. [0015]
  • SUMMARY OF THE INVENTION
  • One object of the present invention is to provide a means for more precisely depositing samples on a sample plate of a mass spectrometry apparatus. [0016]
  • Another object of the present invention is to provide a means for maximizing the number of samples spotted on the surface of a sample plate of a mass spectrometry apparatus. [0017]
  • An aspect of the present invention relates to the use of an alignment plate to assist in accurate positioning of a pipette tip delivering samples onto a sample plate of a mass spectrometer. [0018]
  • Another aspect of the present invention relates to the use of an adaptor plate and an alignment plate with an X-Y manipulator device originally designed for liquid sample manipulation. Use of the adaptor plate and alignment plate of the present invention allows for new usage of the X-Y manipulator device. Specifically, the X-Y manipulator device can be used for spotting samples on a sample plate of a mass spectrometer with the alignment providing alignment holes for accurate positioning of samples on the sample plate. [0019]
  • In accordance with yet another aspect of the present invention, a plurality of alignment plates are used to guide a pipette tip toward a sample plate for accurate positioning of samples on the sample plate. A first alignment plate guides the pipette tip to spot an array of samples on the sample plate. After replacing the first alignment plate with a second alignment plate, a second array of samples can be used to guide the pipette tip to spot a second array of samples on the sample plate, where the second array of samples are located between, but offset from the first array of samples thereby maximizing the number of samples positionable on the sample plate. [0020]
  • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIG. 1 is a side schematic view of a mass spectrometry apparatus; [0021]
  • FIG. 2 is a perspective view of a sample plate for samples to be analyzed in the mass spectrometry apparatus depicted in FIG. 1; [0022]
  • FIG. 3 is a perspective view of an X-Y robotic apparatus used to spot samples on the surface of the sample plate depicted in FIG. 2; [0023]
  • FIG. 4 is top view of an alignment plate having a 10×10 array of alignment holes used for positioning samples on a sample plate in accordance with one embodiment of the present invention; [0024]
  • FIG. 5 is side exploded cross-section view taken along the line V-V in FIG. 4 showing the sample plate mounted on the table of an X-Y robotic apparatus, with the alignment plate depicted in FIG. 4 being mounted thereon in accordance with the present invention; [0025]
  • FIG. 6 is a side cross-section view taken along the line VI-VI in FIG. 4 showing the sample plate mounted on the table of an X-Y robotic apparatus, with the alignment plate depicted in FIGS. 4 and 5 mounted thereon, and a pipette delivering a sample through a hole in the alignment plate to the surface of the sample plate in accordance with the present invention; [0026]
  • FIG. 7 is a top view of the sample plate with an array of 10×10 samples accurately positioned on the upper surface thereof in accordance with the present invention; [0027]
  • FIG. 8 is top view of a second alignment plate having a 9×9 array of alignment holes used for positioning samples on a sample plate in accordance with the present invention, with the 9×9 array of alignment holes being located to compliment the 10×10 array of holes in the alignment plate depicted in FIG. 4; [0028]
  • FIG. 9 is another top view of the sample plate depicted in FIG. 7 with the array of 10×10 samples accurately positioned on the upper surface thereof along with an additional 9×9 array of samples loaded using the alignment plate depicted in FIG. 8 in accordance with the present invention; [0029]
  • FIG. 10 is a top view of an adaptor plate for use with various types of X-Y robotic apparatus in accordance with a second embodiment of the present invention; [0030]
  • FIG. 11 is a top view of an alignment plate in accordance with the second embodiment of the present invention; [0031]
  • FIG. 12 is a side view showing two sample plates mounted on the adaptor plate depicted in FIG. 10, with the alignment plate depicted in FIG. 11 mounted thereon, and a pipette delivering a sample through a hole in the alignment plate to the surface of one of the sample plates in accordance with the present invention; [0032]
  • FIG. 13 is an exploded perspective view showing the alignment plate, one sample plate and the adaptor plate, where the alignment plate is provided with a 10×10 array of alignment holes in accordance with the present invention; [0033]
  • FIG. 14 is an exploded perspective view showing another alignment plate, one sample plate and the adaptor plate, where the alignment plate is provided with a 9×9 array of alignment holes in accordance with the present invention; and [0034]
  • FIG. 15 is a block diagram showing the X-Y robotic apparatus and a computer for controlling the X-Y robotic apparatus in accordance with the present invention.[0035]
  • DETAILED DESCRIPTION OF THE INVENTION
  • The present invention relates to alignment plates that include holes for guiding a pipette tip toward a sample plate thereby assisting in accurately spotting or depositing samples on a sample plate of a mass spectrometry apparatus. FIG. 4 is top view of an [0036] alignment plate 60 that is formed with a plurality of holes 61 that define an array of holes. In the embodiment depicted in FIG. 4, there are one hundred holes 61 defining a 10×10 array of alignment holes used for positioning samples on a sample plate 15 in accordance with one embodiment of the present invention.
  • Each of the [0037] holes 61 extends from an upper surface 62 of the alignment plate 61 to a lower surface 65. At the upper surface 61, each hole 61 is further provided with a conical contour 63 whose function is described in greater detail below. The alignment plate 61 is further formed with a plurality of positioning legs 68 that extend downward away from the upper surface 62.
  • The X-Y robotic apparatus described above with respect to FIG. 3 is typically formed with means for retaining the [0038] sample plate 15. Specifically, the X-Y robotic apparatus includes a table 48 that includes a recess 49 for receiving the sample plate 15, as shown in FIGS. 5 and 6. The recess 49 snuggly receives the sample plate 15 thereby retaining the sample plate 15 in a stationary position. The alignment plate 60 and its positioning legs 68 are formed to conform to the surface contours of the table 48 such that the sample plate 15 may be sandwiched in between the table 48 and the alignment plate 60, as depicted in FIG. 6. The positioning legs 68 firmly engage the contours of the table 48 such that the alignment plate 60 is held in a stationary position above the sample plate 15, as depicted in FIG. 6. Alternatively, the positioning legs 68 may be dimensioned to contact the outer edges of the sample plate 15.
  • FIG. 6 is a side view showing the sample plate mounted on the table of the X-Y robotic apparatus, with the [0039] alignment plate 60 mounted thereon. A pipette tip 45 is shown extending through one of the holes 61 in the alignment plate 60 delivering the sample to the upper surface 62 of the sample plate 60. As described above, the X-Y robotic apparatus depicted in FIG. 3 positions the pipette tip 45, above the sample plate 15 in order to deposit a sample. With the alignment plate 60 positioned above the sample plate 15, as the X-Y robotic apparatus moves the pipette tip 45 downward toward the sample plate 15, the pipette tip 45 engages the conical contour 63 of the hole 61 in the alignment plate 60. As the pipette tip 45 contacts the conical contour 63, it is guided into the hole 61 and toward the upper surface of the sample plate 15. Since the alignment plate 60 is finely machined with the centers of the holes 61 in the 10×10 array at accurate, predetermined distances from one another, the samples are reliably positioned with respect to one another, as shown in FIG. 7. FIG. 7 is a top view of the sample plate 15 with an array of 10×10 samples accurately positioned on the upper surface of the sample plate 15. In other words, the pipette tip 45 is guided into and through the holes 61 in the alignment plate 60 in order to more reliably position the samples on the sample plate 15.
  • It should be understood that the [0040] alignment plate 60 depicted in FIGS. 4 and 5 is not limited to the 10×10 array of holes 61. Alternatively, a greater number of holes may be formed. For instance, the inventors have made an alternate embodiment of the alignment plate that has a 17×17 array of holes (not shown).
  • It should further be appreciated that the X-Y robotic apparatus and the replaceable pipette tips are guided into position by the [0041] conical contour 63 of the holes 61 in order to deposit samples on the sample plate 15. Therefore, positional accuracy of the X-Y robotic apparatus and deformable nature of the pipette tips 45 are no longer so critical because positional accuracy is now provided by the alignment plate 60.
  • FIG. 8 is top view of a [0042] second alignment plate 70 having a 9×9 array of alignment holes used for positioning samples on the sample plate 15 in accordance with an additional feature of the present invention. The 9×9 array of alignment holes 61 are located to compliment the 10×10 array of holes in the alignment plate 60 depicted in FIG. 4. Specifically, after the 10×10 array of samples is deposited on the sample plate 15, as shown in FIG. 7, the alignment plate 60 is removed from the table 48. The second alignment plate 70 is then installed on the table 48 above the sample plate 15 having the 10×10 array of samples (FIG. 7) deposited thereon. An additional eighty-one (81) samples are then deposited on to the sample plate 15 to form the two space apart arrays of samples on the sample plate 15, as shown in FIG. 9. In other words, first the alignment plate 60 is used to accurately position the pipette tip 45 above the sample plate 15 in order to deposit a 10×10 array of samples on the sample plate 15. Thereafter, the second alignment plate 60 is used to accurately position the pipette tip 45 above the sample plate 15 in order to deposit an additional 9×9 array of samples between the 10×10 array of samples, as depicted in FIG. 9.
  • It should be understood that the [0043] second alignment plate 70 can have any number of holes in order to complement the holes in the first alignment plate 60. For instance, if the first alignment plate 60 is provided with a 12×12 array of alignment holes, the second alignment plate 70 is provided with either a 11×11 array of alignment holes or a 13×13 array of alignment holes to significantly increase the number of samples deposited on the sample plate 15. Further, the second alignment plate 70 described above and shown in the drawings includes an array of holes that are configured to be diagonally offset from the holes in the first alignment plate. Alternatively, the array of holes in the second alignment plate may be offset from the holes in the first plate along one axis, not diagonally offset. Further, a series of alignment plates can be configured to assist in spotting three or four separate arrays of spots on a sample plate, with the alignment holes in each alignment plate being offset from the other alignment plate holes.
  • In a second embodiment of the present invention, the [0044] sample plate 15 is loaded with samples by an X-Y robotic apparatus that is not provided with a recess for retaining the sample holder 15. Instead, an adaptor plate 80 depicted in FIG. 10 is used to retain the sample holder 15. Specifically, the adaptor plate 80 has an outer perimeter 82 that is formed with dimensions corresponding to retainer portions 85 (shown in FIGS. 13 and 14) on an X-Y robotic apparatus. The adaptor plate 80 is provided in order to deposit samples on the sample plate 15 using an X-Y robotic apparatus that is not originally manufactured for use with a MALDI-TOF apparatus.
  • FIG. 10 is a top view of the [0045] adaptor plate 80. The adaptor plate 80 is formed with at least one recess 49′ that is shaped to securely receive and retain the sample plate 15. The recess 49′ and the recess 49 described above have generally the same shape. The recess 49′ (and the recess 49 in FIGS. 5 and 6) includes an alignment recess 49 d that receives an alignment pin 15 d such that the sample plate 15 only fits into the recess 49′ in a single orientation.
  • The [0046] adaptor plate 80 is formed with two recesses 49′ but could be formed with more than two recesses 49′ providing the adaptor plate 80 is large enough to receive such recesses. The size of the adaptor plate 80 is dependent upon the size of the retainer portions 85 of the X-Y robotic apparatus, since the adaptor plate 80 is made to be received in the retainer portions 85.
  • A [0047] first alignment plate 90 shown in FIG. 11, is dimensioned to correspond to the size and shape of the adaptor plate 80. Since the adaptor plate 80 is formed with two recesses 49′ to receive and securely retain two sample plates 15, the first alignment plate 90 is formed with two separate 10×10 arrays of alignment holes 91. Each two 10×10 arrays of alignment holes 91 corresponds to one of the sample plates 15. Each alignment hole 91 is formed with a conical contour 92 to guide a pipette as the pipette deposits a sample on the sample plate 15. The first alignment plate 90 is further formed with positioning legs 94 for holding the first alignment plate 90 on the adaptor plate 80.
  • FIG. 12 is a slightly exploded side view showing two [0048] sample plates 15 mounted on the adaptor plate 80, with the first alignment plate 90 depicted in FIG. 11 mounted thereon. A pipette tip 45 is shown extending through one of the alignment holes 91 depositing a sample on surface of one of the sample plates 15.
  • FIG. 13 is an exploded perspective view showing the [0049] first alignment plate 90, one sample plate 15 and the adaptor plate 80, where the first alignment plate is provided with a 10×10 array of alignment holes. In a manner similar to the first embodiment depicted in FIGS. 4-9, a second alignment plate 98 shown in FIG. 14 is provided with a 9×9 array of alignment holes in order to load a second array of samples onto the sample plate 15.
  • It should be understood from the above description that the first and [0050] second alignment plates 90 and 98 allow multiple arrays of samples to be deposited on a single sample plate 15. Further the adaptor plate 80, in combination with the first and second alignment plates 90 and 98 allows for two sample plates 15 to be loaded with multiple arrays of samples.
  • It should also be understood that the specific number of alignment holes in the alignment plates of the present invention is not limited to the specific number shown in the drawings. The alignment plates of the present invention may be provided with an array of holes of over to 20×20 holes if desired and the size of the sample plate allows. The number of holes in any array of holes is dependent primarily upon the size of the sample plate and the size of the samples to be deposited on the sample plate. [0051]
  • Further, it should be understood that the present invention is not limited to the specific sample plate depicted. There are several MALDI apparatus currently available. The present invention is applicable to all. For instance, the sample plate depicted is about 2.25 inches by 2.25 inches in size. Other MALDI apparatus' use sample plates that are larger and can hold more samples. The present invention is applicable to larger sample plates as well. Specifically, the alignment plates described above can be dimensioned to fit any sample plate. [0052]
  • In the field of proteomics, samples to be deposited on the [0053] sample plate 15 can be prepared in a variety of ways. For example, for cellular matter analysis, the samples can be based upon processed spots initially excised from a 2-D electrophoresis gel slab. 2-D electrophoresis gel slabs are prepared in a variety of ways, for instance see U.S. Pat. No. 5,993,627, which is incorporated herein by reference in its entirety. After excising a spot from a 2-D gel, the spot is deposited in a 96-well microtiter plate (aka 96-well plate 50 depicted in FIG. 3) or microcentrifuge tube. The spot may include a specific peptide or protein that is to be subjected to mass spectrometry analysis.
  • After loading one or more excised spots into the 96-[0054] well plate 50 or microcentrifuge tube, the 96-well plate 50 or microcentrifuge tube is then moved to an X-Y robotic apparatus, such as the X-Y robotic apparatus 48 described above with respect to FIG. 3. The X-Y robotic apparatus 48 is typically controlled by a computer 100, as shown in FIG. 15. The computer 100 typically includes an input 105 such as a keyboard, floppy disk drive and or mouse. The computer 100 also includes a display 110 and an output 115.
  • The X-Y [0055] robotic apparatus 48, under the control of the computer 100, performs several procedures on each of the spots in the 96-well plate 50 in order to prepare the excised spots for spotting on the sample plate. For instance, the spot, which includes gel slab material, must be digested in order to remove the unwanted gel material. Such procedures are well known in the art and vary depending upon various circumstances. However, almost all digestion procedures include manipulation of fluid in and about the 96-well plate 50 retaining the samples. There are many X-Y robotic apparatus 48 available commercially for manipulation of such liquids in the preparation of samples.
  • After each sample is digested, and processed for sample preparation, each sample is typically moved to a clean 96-well plate by the X-Y [0056] robotic device 48. The computer 100 records identification of the sample and its location on the sample plate 15 in memory (not shown) and is configured to output location information upon request. In the above described manner, spots excised from a 2-D gel are processed and thereafter deposited on a sample plate 15 for analysis in the MALDI apparatus 1.
  • An advantage of the present invention is that the X-Y [0057] robotic apparatus 48 that performs the liquid manipulation in the sample preparation procedures may now be used to spot the samples on the sample plate 15 using the alignment plates and adaptor plate described above.
  • By virtue of the above described alignment plates it is possible to increase the sample carrying capabilities of a sample plate of a mass spectrometry apparatus. It is further possible to more reliably position the samples on the sample plate for more reliable targeting of the laser of the mass spectrometry apparatus. [0058]
  • By virtue of the above described alignment plates in combination with the adaptor plate, it is possible to increase the sample carrying capabilities of a sample plate of a mass spectrometry apparatus using an X-Y robotic device that is not specifically designed for mass spectrometry sample plate loading for depositing samples on a sample plate. Specifically, using the alignment plates and adaptor plate of the present invention, it is possible to augment a liquid handling X-Y robotic device such that the X-Y robotic device may be used for unintended use, thereby broadening the usefulness of the X-Y robotic device. [0059]
  • While the invention has been described in detail above, the invention is not intended to be limited to the specific embodiments as described. It is evident that those skilled in the art may now make numerous uses and modifications of and departures from the specific embodiments described herein without departing from the inventive concepts. [0060]

Claims (9)

What is claimed is:
1. An alignment plate for depositing samples on a sample plate, the alignment plate comprising:
a plate member, said plate member being formed with a plurality of alignment holes extending from one side of said plate member to another side of said plate member, each of said alignment holes being formed with a conical contour at said one side of said plate member for guiding a pipette tip therethrough.
2. An alignment plate as set forth in claim 1, further comprising:
a plurality of positioning legs extending away from said one side, said positioning legs being provided to align said plate member with the sample plate.
3. An alignment plate as set forth in claim 2, wherein said positioning legs are formed to engage and align with an adaptor plate configured to receive and retain the sample plate.
4. An alignment plate as set forth in claim 2, wherein said plurality of alignment holes comprises a 10×10 array of holes.
5. An alignment plate as set forth in claim 2, wherein said plurality of alignment holes comprises a 17×17 array of holes.
6. An alignment plate as set forth in claim 2, wherein said plurality of alignment holes comprises a 9×9 array of holes.
8. An alignment plate as set forth in claim 2, wherein the sample plate is part of a MALDI mass spectrometer.
9. An alignment apparatus comprising:
a support member;
a mass spectrometer sample plate removably coupled to said support member; and
an alignment plate removably coupled to said support member and being oriented above and substantially parallel to said sample plate, said alignment plate having a top surface, a bottom surface and a plurality of spaced-apart alignment holes extending between said top surface and said bottom surface, said alignment holes being oriented in an array and spaced-apart a distance corresponding to a predetermined spacing of samples on said sample plate, each of said alignment holes having a dimension to enable a pipette to pass through said alignment plate for pipetting a sample onto said sample plate.
10. The alignment apparatus of claim 9, wherein said alignment plate includes a plurality of legs extending from said bottom surface and engaging said support member.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7109481B1 (en) * 2005-04-28 2006-09-19 Thermo Finnigan Llc Matrix-assisted laser desorption and ionization (MALDI) sample plate releasably coupled to a sample plate adapter
US20130146758A1 (en) * 2010-05-21 2013-06-13 Empa Eidg. Materialprufungs-Und Forschungsanstalt High-density sample support plate for automated sample aliquoting
US9588133B2 (en) 2011-12-21 2017-03-07 Roche Molecular Systems, Inc. Analytical system with tip rack assembly configured to prevent contamination
US10690626B2 (en) * 2014-01-17 2020-06-23 Coastal Genomics Inc. Cassettes for use in automated parallel electrophoretic assays and methods for manufacturing and using same
JPWO2019058783A1 (en) * 2017-09-21 2020-09-03 浜松ホトニクス株式会社 Sample support

Families Citing this family (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6508986B1 (en) * 1999-08-27 2003-01-21 Large Scale Proteomics Corp. Devices for use in MALDI mass spectrometry
EP1358480A2 (en) * 2001-02-01 2003-11-05 The Johns Hopkins University Diagnosis of pathogen infections using mass spectral analysis of immune system modulators in post-exposure biological samples.
US20040234971A1 (en) * 2001-02-01 2004-11-25 Joany Jackman Diagnosis of pathogen infections using mass spectral analysis of immune system modulators in post-exposure biological samples
US7215813B2 (en) 2001-12-03 2007-05-08 Apple Computer, Inc. Method and apparatus for color correction
US7332347B2 (en) * 2003-04-14 2008-02-19 Liang Li Apparatus and method for concentrating and collecting analytes from a flowing liquid stream
US7138625B2 (en) * 2003-05-02 2006-11-21 Agilent Technologies, Inc. User customizable plate handling for MALDI mass spectrometry
US20040241659A1 (en) * 2003-05-30 2004-12-02 Applera Corporation Apparatus and method for hybridization and SPR detection
US7145135B1 (en) * 2003-05-30 2006-12-05 Agilent Technologies, Inc. Apparatus and method for MALDI source control with external image capture
US6825478B1 (en) * 2003-10-10 2004-11-30 Perseptive Biosystems, Inc. MALDI plate with removable magnetic insert
US7095018B2 (en) * 2004-12-29 2006-08-22 Wisconsin Alumni Research Foundation Deposition of samples and sample matrix for enhancing the sensitivity of matrix assisted laser desorption/ionization mass spectrometry
US7435951B2 (en) * 2005-06-08 2008-10-14 Agilent Technologies, Inc. Ion source sample plate illumination system
US7495231B2 (en) * 2005-09-08 2009-02-24 Agilent Technologies, Inc. MALDI sample plate imaging workstation
US8105788B2 (en) * 2005-12-08 2012-01-31 The University Of British Columbia Mucopolysaccharidosis (MPS) diagnostic methods, systems, kits and assays associated therewith
GB0717462D0 (en) * 2007-09-07 2007-10-17 Mole Genetics As Liquid delivery apparatus
DE102016124017B3 (en) * 2016-12-12 2017-12-28 Bruker Daltonik Gmbh Apparatus and method for preparing samples for ionization by laser desorption in a mass spectrometer
CN106960777B (en) * 2016-12-31 2019-08-20 宁波华仪宁创智能科技有限公司 Mass spectrometry system and its working method
GB2582135B (en) * 2019-03-11 2023-06-21 Kratos Analytical Ltd Mass spectrometry apparatus
US11958053B2 (en) * 2021-09-21 2024-04-16 The Government of the United States of America, as represented by the Secretary of Homeland Security Media holder for sample preparation

Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4699884A (en) * 1984-02-29 1987-10-13 Gerhard Noss Process and apparatus for the simultaneous application of a multiplicity of liquid samples to an object stage
US5262128A (en) * 1989-10-23 1993-11-16 The United States Of America As Represented By The Department Of Health And Human Services Array-type multiple cell injector
US5443791A (en) * 1990-04-06 1995-08-22 Perkin Elmer - Applied Biosystems Division Automated molecular biology laboratory
US5487997A (en) * 1994-02-15 1996-01-30 Point Plastics Incorporated Pipette tip mounting and transfer apparatus and method
US5719060A (en) * 1993-05-28 1998-02-17 Baylor College Of Medicine Method and apparatus for desorption and ionization of analytes
US5770860A (en) * 1996-07-12 1998-06-23 Franzen; Jochen Method for loading sample supports for mass spectrometers
US5882930A (en) * 1997-11-10 1999-03-16 Hyseq, Inc. Reagent transfer device
US5888831A (en) * 1997-03-05 1999-03-30 Gautsch; James W. Liquid-sample-separation laboratory device and method particularly permitting ready extraction by syringe of the separated liquid sample
US5993627A (en) * 1997-06-24 1999-11-30 Large Scale Biology Corporation Automated system for two-dimensional electrophoresis
US6024925A (en) * 1997-01-23 2000-02-15 Sequenom, Inc. Systems and methods for preparing low volume analyte array elements
US6132582A (en) * 1998-09-14 2000-10-17 The Perkin-Elmer Corporation Sample handling system for a multi-channel capillary electrophoresis device
US6175112B1 (en) * 1998-05-22 2001-01-16 Northeastern University On-line liquid sample deposition interface for matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy
US6267930B1 (en) * 1997-09-22 2001-07-31 Waldemar Ruediger Apparatus for synthesis of multiple organic compounds with pinch valve block
US6287872B1 (en) * 1997-12-11 2001-09-11 Bruker Daltonik Gmbh Sample support plates for Maldi mass spectrometry including methods for manufacture of plates and application of sample
US6302159B1 (en) * 1999-06-09 2001-10-16 Genomic Solutions, Inc. Apparatus and method for carrying out flow through chemistry of multiple mixtures
US6362004B1 (en) * 1999-11-09 2002-03-26 Packard Biochip Technologies, Llc Apparatus and method for using fiducial marks on a microarray substrate
US6508986B1 (en) * 1999-08-27 2003-01-21 Large Scale Proteomics Corp. Devices for use in MALDI mass spectrometry

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5011779A (en) * 1988-01-21 1991-04-30 Long Island Jewish Medical Center Apparatus for rapid deposition of test samples on an absorbent support
EP0527726A1 (en) * 1990-05-08 1993-02-24 Wallac Oy An apparatus for counting liquid scintillation samples
DE19712195B4 (en) * 1997-03-22 2007-12-27 Friedrich-Schiller-Universität Jena Method and apparatus for providing samples for off-line detection of analytes according to MALDI mass spectrometry
US6627446B1 (en) * 1998-07-02 2003-09-30 Amersham Biosciences (Sv) Corp Robotic microchannel bioanalytical instrument

Patent Citations (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4699884A (en) * 1984-02-29 1987-10-13 Gerhard Noss Process and apparatus for the simultaneous application of a multiplicity of liquid samples to an object stage
US5262128A (en) * 1989-10-23 1993-11-16 The United States Of America As Represented By The Department Of Health And Human Services Array-type multiple cell injector
US5443791A (en) * 1990-04-06 1995-08-22 Perkin Elmer - Applied Biosystems Division Automated molecular biology laboratory
US5719060A (en) * 1993-05-28 1998-02-17 Baylor College Of Medicine Method and apparatus for desorption and ionization of analytes
US6124137A (en) * 1993-05-28 2000-09-26 Baylor College Of Medicine Surface-enhanced photolabile attachment and release for desorption and detection of analytes
US5487997A (en) * 1994-02-15 1996-01-30 Point Plastics Incorporated Pipette tip mounting and transfer apparatus and method
US5770860A (en) * 1996-07-12 1998-06-23 Franzen; Jochen Method for loading sample supports for mass spectrometers
US6024925A (en) * 1997-01-23 2000-02-15 Sequenom, Inc. Systems and methods for preparing low volume analyte array elements
US5888831A (en) * 1997-03-05 1999-03-30 Gautsch; James W. Liquid-sample-separation laboratory device and method particularly permitting ready extraction by syringe of the separated liquid sample
US5993627A (en) * 1997-06-24 1999-11-30 Large Scale Biology Corporation Automated system for two-dimensional electrophoresis
US6267930B1 (en) * 1997-09-22 2001-07-31 Waldemar Ruediger Apparatus for synthesis of multiple organic compounds with pinch valve block
US5882930A (en) * 1997-11-10 1999-03-16 Hyseq, Inc. Reagent transfer device
US6287872B1 (en) * 1997-12-11 2001-09-11 Bruker Daltonik Gmbh Sample support plates for Maldi mass spectrometry including methods for manufacture of plates and application of sample
US6175112B1 (en) * 1998-05-22 2001-01-16 Northeastern University On-line liquid sample deposition interface for matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy
US6132582A (en) * 1998-09-14 2000-10-17 The Perkin-Elmer Corporation Sample handling system for a multi-channel capillary electrophoresis device
US6302159B1 (en) * 1999-06-09 2001-10-16 Genomic Solutions, Inc. Apparatus and method for carrying out flow through chemistry of multiple mixtures
US6508986B1 (en) * 1999-08-27 2003-01-21 Large Scale Proteomics Corp. Devices for use in MALDI mass spectrometry
US6362004B1 (en) * 1999-11-09 2002-03-26 Packard Biochip Technologies, Llc Apparatus and method for using fiducial marks on a microarray substrate

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7109481B1 (en) * 2005-04-28 2006-09-19 Thermo Finnigan Llc Matrix-assisted laser desorption and ionization (MALDI) sample plate releasably coupled to a sample plate adapter
US20130146758A1 (en) * 2010-05-21 2013-06-13 Empa Eidg. Materialprufungs-Und Forschungsanstalt High-density sample support plate for automated sample aliquoting
US9211542B2 (en) * 2010-05-21 2015-12-15 Eidgenossische Technische Hochschule Zurich High-density sample support plate for automated sample aliquoting
US9588133B2 (en) 2011-12-21 2017-03-07 Roche Molecular Systems, Inc. Analytical system with tip rack assembly configured to prevent contamination
US10101354B2 (en) 2011-12-21 2018-10-16 Roche Molecular Systems, Inc. Method for prevention of contamination during disposing of liquid by pipet array
US10690626B2 (en) * 2014-01-17 2020-06-23 Coastal Genomics Inc. Cassettes for use in automated parallel electrophoretic assays and methods for manufacturing and using same
JPWO2019058783A1 (en) * 2017-09-21 2020-09-03 浜松ホトニクス株式会社 Sample support
JP7026121B2 (en) 2017-09-21 2022-02-25 浜松ホトニクス株式会社 Sample support
US11355333B2 (en) 2017-09-21 2022-06-07 Hamamatsu Photonics K.K. Sample support body

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US6508986B1 (en) 2003-01-21

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