LED point scanning system

• US 5,578,818 A
• Filed: 05/10/1995
• Issued: 11/26/1996
• Est. Priority Date: 05/10/1995
• Status: Expired due to Term

• Current Assignee: GE Healthcare Bio-Sciences Corporation (General Electric Company)
• Original Assignee: Molecular Dynamics Limited
• Inventors: Majlof, Lars, Kain, Robert C., Miller, Michael F.
• Primary Examiner(s): Allen, Stephone
• Application Number:
  US08/438,416
• Time in Patent Office: 566 Days
• Field of Search: 250/201.3, 250/234-235, 250/458.1, 250/459.1, 250/461.1, 250/461.2, 356/318, 356/326, 356/328
• US Class Current: 250/234
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates

Abstract

An LED point imaging scanner for stimulating and for reading fluorescent and reflective signal radiation from a target sample is disclosed. The LED light source is mounted on a scan head and is focused into the pinhole aperture of a spatial filter, then collimated and focused to a spot on the sample. The spatial filter in the illumination path, or an optical fiber serving the equivalent function, effectively restricts the incoherent LED light and creates a point light source. Signal radiation from the sample is collected through the scan head and the returning light beam is collimated and focused onto the receiving section of a detection means. A spatial filter in the path of the returning beam may also be utilized. The detection means is either a small detector which is mounted directly on the scan head or an end of an optical fiber which transmits the point image to a remotely-positioned stationary detector. The scan head is moved along one or two axes relative to the sample for scanning of a target area of the sample.

Claims

1. An LED optical system for point scanning of a sample comprising:

(a) a movable scan head having

(i) an LED light source providing an illumination beam along a single path,

(ii) an illumination spatial filter having a pinhole aperture spaced apart from the light source,

(iii) means for focusing the illumination beam into the pinhole aperture,

(iv) means for directing the illumination beam after the illumination beam has been focused into the pinhole aperture onto a spot of the sample to cause signal radiation to be returned from the sample, and

(v) means for collecting and detecting the signal radiation returned from the sample,

(b) a stage for supporting the sample, and

(c) means for moving the scan head along a first axis relative to the stage to illuminate a plurality of spots along a scan line on the sample.

2. The system of claim 1 further comprising:

means for moving the scan head along a second axis relative to the stage to cause illumination of spots on a plurality of parallel scan lines on the sample, the second axis being perpendicular to the first axis.

3. The system of claim 1 wherein the means for directing the illumination beam onto a spot of the sample comprises:

a collimating lens for receiving and collimating the illumination beam after the illumination beam has been focused into the pinhole aperture, and

an objective lens for focusing the collimated illumination beam onto a spot of the sample.

4. The system of claim 3 wherein the collimating lens has a numerical aperture and the objective lens has a numerical aperture, the numerical apertures of the collimating lens and the objective lens being equal.

5. The system of claim 1 wherein the means for collecting and detecting the signal radiation returned from the sample comprises:

an objective lens for directing the signal radiation to form a collimated returning light beam, and

a focusing lens for receiving at least a portion of the collimated returning light beam and focusing the portion into an optical fiber, the fiber transmitting the returning light beam to a stationary detector removed from the movable scan head.

6. The system of claim 5 further comprising a beam splitter disposed in the path of the returning light beam, wherein the beam splitter comprises a wavelength discrimination filter.

7. The system of claim 5 further comprising a beam splitter disposed in the path of the returning light beam, wherein the beam splitter comprises a partial reflection filter.

8. The system of claim 1 wherein the means for collecting and detecting the signal radiation returned from the sample comprises:

an objective lens for directing the signal radiation to form a collimated returning light beam, and

a detector for receiving at least a portion of the returning light beam, the detector being mounted on the movable scan head.

9. The system of claim 8 further comprising:

a focusing lens, disposed between the objective lens and the detector, for receiving at least a portion of the collimated returning light beam and focusing the portion onto the detector.

10. The system of claim 8 further comprising:

a detection spatial filter having a pinhole aperture, disposed between the objective lens and the detector, and

a focusing lens, disposed between the objective lens and the detection spatial filter, for receiving at least a portion of the collimated returning light beam and focusing the portion into the pinhole aperture of the detection spatial filter.

11. The system of claim 8 further comprising a focusing lens, disposed between the objective lens and the detector, for receiving at least a portion of the collimated returning light beam and focusing the portion into an optical fiber, the fiber transmitting the returning light beam toward the detector.

12. The system of claim 8 further comprising a beam splitter disposed in the path of the returning light beam, wherein the beam splitter comprises a wavelength discrimination filter.

13. The system of claim 8 further comprising a beam splitter disposed in the path of the returning light beam, wherein the beam splitter comprises a partial reflection filter.

14. An LED optical system for point scanning of a sample comprising:

(a) an LED light source providing an illumination beam along a single path,

(b) means for focusing the illumination beam into an optical fiber, the fiber transmitting the illumination beam,

(c) a movable scan head having

(i) means for receiving the transmitted illumination beam from the fiber and for directing the illumination beam onto a spot of the sample to cause signal radiation to be returned from the sample, and

(ii) means for collecting and detecting the signal radiation returned from the sample,

(d) a stage for supporting the sample, and

(e) means for moving the scan head along a first axis relative to the stage to illuminate a plurality of spots along a scan line on the sample.

15. The system of claim 14 further comprising:

means for moving the scan head along a second axis relative to the stage to cause illumination of spots on a plurality of parallel scan lines on the sample, the second axis being perpendicular to the first axis.

16. The system of claim 14 wherein the LED light source and the means for focusing the illumination beam into an optical fiber are mounted on the movable scan head.

17. The system of claim 14 wherein the means for collecting and detecting the signal radiation returned from the sample comprises:

an objective lens for directing the signal radiation to form a collimated returning light beam, and

a focusing lens for receiving at least a portion of the collimated returning light beam and focusing the portion into an optical fiber, the fiber transmitting the returning light beam to a stationary detector removed from the movable scan head.

18. The system of claim 14 wherein the means for collecting and detecting the signal radiation returned from the sample comprises:

an objective lens for directing the signal radiation to form a collimated returning light beam, and

a detector for receiving at least a portion of the returning light beam, the detector being mounted on the movable scan head.

19. A moving scan head for point imaging of a sample, the scan head comprising:

an LED source providing an illumination beam along a single path,

an illumination spatial filter having a pinhole aperture,

a first focusing lens disposed between the LED source and the illumination spatial filter for focusing the illumination beam into the pinhole aperture of the illumination spatial filter,

a collimating lens for collimating the illumination beam after the illumination beam has been focused into the pinhole aperture,

a beam splitter disposed in the illumination beam path for reflecting at least a portion of the collimated illumination beam,

an objective lens for receiving the portion of the illumination beam from the beam splitter and for focusing the portion of the illumination beam onto a spot of the sample to cause signal radiation to emerge from the sample, and for receiving and collimating the signal radiation from the sample to form a collimated returning beam, the beam splitter also disposed in the returning beam path for passing at least a portion of the collimated returning beam, and

a receiving section of a detection means disposed to intercept the returning beam.

20. The scan head of claim 19 further comprising:

a second focusing lens for receiving the portion of the collimated returning beam that has passed through the beam splitter and for focusing the portion of the collimated returning beam onto the receiving section of the detection means.

21. The scan head of claim 19 further comprising:

a detection spatial filter having a pinhole aperture disposed in the path of the returning beam, and

a second focusing lens for receiving the portion of the collimated returning beam that has passed through the beam splitter and for focusing the portion of the collimated returning beam into the pinhole aperture of the detection spatial filter.

22. The scan head of claim 19 wherein the collimating lens has a numerical aperture and the objective lens has a numerical aperture, the numerical apertures of the collimating lens and the objective lens being equal.

23. The scan head of claim 19 wherein the beam splitter disposed in the illumination beam path for reflecting at least a portion of the collimated illumination beam and disposed in the returning beam path for passing at least a portion of the collimating returning beam comprises a wavelength discrimination filter.

24. The scan head of claim 19 wherein the beam splitter disposed in the illumination beam path for reflecting at least a portion of the collimated illumination beam and disposed in the returning beam path for passing at least a portion of the collimating returning beam comprises a partial reflection filter.

25. The scan head of claim 19 wherein the detection means is a photomultiplier tube mounted on the scan head.

26. The scan head of claim 19 wherein the detection means is a solid state detector mounted on the scan head.

27. The scan head of claim 19 wherein the receiving section of the detection means is an end of an optical fiber, the fiber transmitting the returning beam to the detection means.

28. An LED point scanner for imaging of a sample, the scanner comprising:

(a) an incident beam means having

(i) a movable light source providing an illumination beam from an LED along a single path,

(ii) a spatial filter having a pinhole aperture,

(iii) means for focusing the illumination beam into the pinhole aperture,

(iv) means for collimating the illumination beam after the illumination beam has been focused into the pinhole aperture, and

(v) means for receiving the collimated illumination beam and for focusing the collimated illumination beam onto a spot of the sample to cause signal radiation to be returned from the sample,

(b) an outgoing beam means having

(i) means for collecting and collimating the signal radiation from the sample into a returning beam, and

(ii) means for focusing the collimated returning beam onto a receiving section of a detection means, and

(c) means for moving the incident and outgoing beam means to provide for scanning along a first axis relative to the sample.

29. The scanner of claim 28 further comprising:

means for moving the incident and outgoing beam means to provide for scanning along a second axis relative to the sample, the second axis being perpendicular to the first axis.

30. The scanner of claim 28 wherein the receiving section of the detection means is a window of a photomultiplier tube, the photomultiplier tube being mounted on the scan head.

31. The scanner of claim 28 wherein the receiving section of the detection means is a window of a solid state detector, the solid state detector being mounted on the scan head.

32. The scanner of claim 28 wherein the receiving section of the detection means is an end of an optical fiber, the fiber transmitting the returning beam to a stationary detector.

33. The scanner of claim 28 further comprising a beam splitter included in the incident and outgoing beam means, wherein the beam splitter comprises a wavelength discrimination filter.

34. The scanner of claim 28 further comprising a beam splitter included in the incident and outgoing beam means, wherein the beam splitter comprises a partial reflection filter.