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WO2000005739A1 - Instrument mecanique electro-optique - Google Patents

Instrument mecanique electro-optique Download PDF

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Publication number
WO2000005739A1
WO2000005739A1 PCT/US1999/016412 US9916412W WO0005739A1 WO 2000005739 A1 WO2000005739 A1 WO 2000005739A1 US 9916412 W US9916412 W US 9916412W WO 0005739 A1 WO0005739 A1 WO 0005739A1
Authority
WO
WIPO (PCT)
Prior art keywords
optical signal
specimen
optical
path
drive mechanism
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US1999/016412
Other languages
English (en)
Inventor
Herman Deweerd
Michael Beach
Jose Hernandez
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Virtek Vision Corp
Original Assignee
Virtek Vision Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Virtek Vision Corp filed Critical Virtek Vision Corp
Priority to EP99935743A priority Critical patent/EP1108262A1/fr
Priority to CA002337830A priority patent/CA2337830A1/fr
Priority to AU51153/99A priority patent/AU5115399A/en
Publication of WO2000005739A1 publication Critical patent/WO2000005739A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052—Optical details of the image generation
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0036—Scanning details, e.g. scanning stages
    • G02B21/0048—Scanning details, e.g. scanning stages scanning mirrors, e.g. rotating or galvanomirrors, MEMS mirrors
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052—Optical details of the image generation
    • G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence

Definitions

  • the present invention relates to optical scanners, and more particularly to a quasi con-focal microscope scanner in which the specimen and the scanner are simultaneously moved relative to each other.
  • Micro array biochips are currently being developed by several biotechnology companies. Micro array biochips are small substrates containing thousands of DNA sequences that represent the genetic codes of a variety of living organisms including human, plant, animal, and pathogens.
  • Biochip technology is used for genetic expression, DNA sequencing of genes, food and water testing for harmful pathogens, and diagnostic screening. Biochips may be used in pharmacogenomics and proteomics research aimed at high throughput screening for drug discovery. High-speed automated biochemistry may lead to drugs for treating illnesses including HIV, cancer, heart disease and others.
  • DNA sequences are extracted from a sample and are tagged with a fluorescent probe, a molecule that, when "excited” by a laser, will emit light of various colors. These fluorescently tagged DNA sequences are then spread over the chip. A DNA sequence will bind to its complementary (cDNA) sequence at a given array location.
  • a typical biochip contains a two- dimensional array of thousands of cDNA sequences, each one unique to a specific gene. These cDNA sequences may be "printed" on the chip in several ways. Once the biochip is printed, it represents thousands of experiments in an area usually smaller than a postage stamp.
  • the chip is then ready to be scanned and analyzed with a scanning laser microscope using a dichromic beam splitter.
  • the dichromic beam splitter has two drawbacks. Each time a specimen with a different dye is to be read, the beam splitter must be changed to match the different wavelengths of operation of the new dye and the number of multiple dyes that can be simultaneously interrogated is usually limited to two.
  • the microscope collects data from successive "pixels" which are best dimensioned in microns.
  • optical scanner There are essentially two types of optical scanner, namely scanners that move scan heads and associated optics over stationary specimens, and scanners that move the specimens relative to stationary optics.
  • Known scanning microscopes must therefore precisely align the optics of a moving scan head with the beam of a stationary laser, or alternatively carry the laser on the moving scan.
  • a stationary laser can be aligned with a moving scan head only at relatively slow speeds, and therefore the scan speed of the system is inherently limited.
  • the alternate system requires a relatively large scan head to carry the associated optics whereby the relatively great size and weight also effectively limits the scan speed.
  • the present invention provides a scanning laser microscope which can be used to scan biochips and display the information embodied in the fluorescent energy emitted by the individual dots as a pictorial representation of the array on a T.V. monitor.
  • the means of interrogation is laser light (the excitation energy).
  • the laser light excites the fluorescein that is contained in the fluorescent dyes.
  • the fluorophores will subsequently emit light of a wavelength that is longer than the wavelength of the excitation energy.
  • the optical diagram is a quasi con-focal microscope, i.e.
  • the size of approximately one pixel is illuminated (excited) and observed (detected) at a time, however the size of the illuminating spot is not nearly as closely matched to that of the detected spot as it is in a pure con-focal microscope, in fact the former is about 10X larger in diameter than the latter.
  • the emitted light is conducted by lenses, mirrors, and optical filters to a detector, where it is converted into computer readable data.
  • the horizontal, or line scan (the X scan) is mechanized by moving the objective lens of the system rapidly back and forth in the X direction across the shorter length of a microscope slide specimen collecting data in each direction.
  • the slide specimen does not move in the X direction as the vertical, or page scan (the Y scan) is mechanized by moving the slide specimen in the Y direction, incrementally advancing the slide each time the X scan is about to start a line.
  • the information is preferably processed so that it may be displayed in a convenient format such as tables, histograms and the like.
  • the pictorial or image-processed information can thereafter be stored on a hard drive and sent to a hard copy printer, transmitted to a LAN, or transmitted over the Internet.
  • Figure 1 is a detailed perspective view of an optical instrument of the present invention
  • Figure 2 is a plan view of a slide specimen of the present invention showing the movement of the scanning objective lens
  • Figure 3 is a side view of the first drive mechanism; and Figure 4 is a top view of the second drive mechanism.
  • optical instrument 10 of the present invention is generally shown in Figure 1.
  • the optical instrument 10 generally includes a transmitter 12 that emits an optical signal 14, a beam splitting mirror 20 having an opening 22, a reflector assembly 30 which directs the optical signal 14 onto a specimen 90, a detector assembly 40 which detects a reflected optical signal 44 from the specimen 90, a first drive mechanism 50 for varying the position of the optical signal 14 on the specimen
  • Figure 1 illustrates the main components of the optical instrument 10 and the optical signal 14 path.
  • the means of interrogation is preferably laser light and more than one laser can be incorporated into the system. Further, various types of lasers may be employed, such as argon-ion, semiconductor diode, and other similar solid state lasers.
  • a plurality of lasers 12A-C each operating on a different wavelength, are shown.
  • the optical signals 14A-C are each first transmitted through a beam correcting lens 16A-C and then through a continuously variable neutral density filter 18A-C, which is employed to adjust the intensity of the beam.
  • the variable neutral density filter 18A-C can be an addressable array of several fixed neutral density filters of different densities, a pair of polarizers of which one is rotatable, or a rotating polarization retarder, in front of a polarizer.
  • the reflector assembly 30 includes a plurality of turn mirrors 32A-C.
  • Each optical signal 14A-C is folded as appropriate by the turn mirrors 32A-C to a beam combiner 34A-C.
  • the beam combiner is preferably a know dichroic filter which transmits light of one wavelength while blocking others.
  • the individual optical signal are thereby collected into a combined beam along a first path which then passes through the opening 22 in the beam splitting mirror 20.
  • the combined beam is then directed to a 90 degree fold mirror 36 located immediately above the scanning objective lens 52.
  • the fold mirror 36 reflects the combined optical signal 14 into the scanning objective lens 52, which in turn is focused onto the specimen 90, thus creating a scanning illumination spot.
  • the embodiment shown in Figure 1 shows three laser transmitters 12A-C, however, those skilled in the art will realize that additional lasers can be used to interrogate multiple dyes of different fluorescent properties in the specimen 90 simultaneously or sequentially.
  • the objective lens 52 preferably outputs a beam of emitted energy concentric with the laser beam, having a diameter about 10X larger than that of the laser beam.
  • the fold mirror 36 located above the scanning lens 52 will fold the reflected optical signal 44 along a second path.
  • the reflected optical signal 44 is again directed by 90 degrees towards the beam splitting mirror 20.
  • the latter will fold the emission beam 90 degrees away from the combined optical signal first path, except for a very small central portion in the middle as determined by the opening 22 in the beam splitting mirror 20. It can be seen that a for a portion of the path the original combined optical signal 14 traveling along the first path, and the reflected optical signal 44 traveling along the second path, have a common path segment. This common path segment is shown between the beam splitting mirror 20 the fold mirror 36, and the scanning lens 52.
  • the reflected optical signal 44 reflecting from the opposite side of the beam splitting mirror 20 will then pass through a plurality of beam splitters 38A-B to separate the combined signal into individual signals 44A-C.
  • Each individual signal 44A-C passes through an emission filter 46A-C, and will then be focused by a detector lens 48A-C into a pinhole.
  • the pinhole acts as the field stop of the system, i.e. , it defines the size of the scanning detection aperture on the slide.
  • the individual signals 46A-C diverts once through the pinhole until it impinges onto a detector 42A-C.
  • the horizontal, or line scan (the X scan) is mechanized by moving the objective lens 52 of the system rapidly (20 Hz or so) back and forth in the X direction across the shorter length of a microscope slide specimen 90 (commonly 1 inch wide), collecting data in each direction.
  • FIG. 3 shows the first drive mechanism 50 for varying the position of the combined optical signal on the specimen 90.
  • the first or X scan mechanism preferably employs a galvanometric torque motor 54 to rotate a sector-shaped cam 56 over an angle between +40 degrees, and -40 degrees.
  • the circular portion of the cam 56 is connected to the carriage 58 via a set of roll-up, roll-off thin, high-strength steel wires 66A-B.
  • the scanning objective lens 52 is attached to the carriage 54.
  • FIG. 4 shows the second drive mechanism 70.
  • the second or Y scan mechanism employs a stepper motor 72 to drive a precision screw 74 in a known manner.
  • the nut 76 on the screw 74 is attached to the carriage 58, so that any rotation of the screw 74 will cause the carriage 58 to move along a linear rail 60.
  • the carriage 58 in turn is equipped with a tray 76.
  • the tray 76 is equipped with appropriate retainers 78 to hold a specimen slide 90 in a position and orientation which is repeatable within an accuracy required by optical focus and alignment criteria.
  • the rail of the linear slide and the stepper motor 72 are attached to the frame of the Y scan mechanism.
  • the carriage 58 is pivotally mounted such that the carriage 58, and thus the objective lens 52, move in an arcuate motion.
  • the arcuate motion is thereafter converted into linear motion by know computer mapping programs.
  • the frame of the Y scan mechanism is further attached to the carriage of a vertically oriented linear slide.
  • the rail of the slide is mounted to the main frame of the reader system.
  • the carriage is supported by a precision screw, the nut of which is attached to the frame. The screw is turned causing the Y scan mechanism, and with it the slide holding the specimen, to move toward or away from the objective lens, thus affecting a focusing sequence.

Landscapes

  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

L'invention concerne un microscope laser à balayage que l'on peut utiliser pour scanner des biopuces et qui comprend un émetteur, y compris des lasers (12A-C), qui émet un signal optique (14), un miroir diviseur de faisceau (20) comportant une ouverture (22), un réflecteur (36) qui dirige un signal optique (14) sur un échantillon (90), un ensemble détecteur, y compris les détecteurs (42A-C), qui détecte un signal optique (44) réfléchi depuis l'échantillon (90), un premier mécanisme d'entraînement destiné à modifier la position du signal optique (14) sur l'échantillon (90) et un deuxième mécanisme d'entraînement destiné à modifier la position de l'échantillon (90) par rapport au signal optique (14).
PCT/US1999/016412 1998-07-23 1999-07-20 Instrument mecanique electro-optique Ceased WO2000005739A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP99935743A EP1108262A1 (fr) 1998-07-23 1999-07-20 Instrument mecanique electro-optique
CA002337830A CA2337830A1 (fr) 1998-07-23 1999-07-20 Instrument mecanique electro-optique
AU51153/99A AU5115399A (en) 1998-07-23 1999-07-20 Electro-optical mechanical instrument

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US9388298P 1998-07-23 1998-07-23
US60/093,882 1998-07-23

Publications (1)

Publication Number Publication Date
WO2000005739A1 true WO2000005739A1 (fr) 2000-02-03

Family

ID=22241515

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1999/016412 Ceased WO2000005739A1 (fr) 1998-07-23 1999-07-20 Instrument mecanique electro-optique

Country Status (4)

Country Link
EP (1) EP1108262A1 (fr)
AU (1) AU5115399A (fr)
CA (1) CA2337830A1 (fr)
WO (1) WO2000005739A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6899700B2 (en) 2001-08-29 2005-05-31 Kimberly-Clark Worldwide, Inc. Therapeutic agent delivery tampon

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5736257A (en) * 1995-04-25 1998-04-07 Us Navy Photoactivatable polymers for producing patterned biomolecular assemblies

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5736257A (en) * 1995-04-25 1998-04-07 Us Navy Photoactivatable polymers for producing patterned biomolecular assemblies
US5847019A (en) * 1995-04-25 1998-12-08 The United States Of America As Represented By The Secretary Of The Navy Photoactivatable polymers for producing patterned biomolecular assemblies

Also Published As

Publication number Publication date
EP1108262A1 (fr) 2001-06-20
CA2337830A1 (fr) 2000-02-03
AU5115399A (en) 2000-02-14

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