Laser excited confocal microscope fluorescence scanner and method

Claim 2

An improved gel scanner comprising means for supporting a gel to be scanned, means for forming a light beam of predetermined wavelength; a dichroic beam splitter for receiving and directing said beam toward a supported gel to be scanned; an objective lens for receiving said beam and focusing the beam in a selected volume of gel to cause fluorescence emission of light at a different wavelength and collecting the emitted light from samples in said selected volume and directing the emitted light to said dichroic beam splitter which passes said emitted light of different wavelengths and reflects light at said predetermined wavelength; a spatial filter for receiving and passing the emitted light from said volume while rejecting background and scattered light; means for providing relative movement between said focused beam and the gel being scanned, means for detecting said passed, emitted light and providing output signals, and means for processing said signals to provide an image of the fluorescence from the gel.

Claim 5

The method of claim 2 in which the excited volume is scanned across the gel to obtain emission from adjacent volumes to provide an output signal for each of said volumes, and processing the output signals to provide an image of the fluorescence from said fragments.

Claim 6

A scanner for scanning a gel to detect fluorescence from fluorescence labeled separated sample components comprising means for supporting a gel to be scanned, means for applying excitation light at a predetermined wavelength to a volume in the gel to cause labeled, separated sample components to fluoresce at a different wavelength, and for collecting the fluorescence emitted light from said sample at said different wavelength from said volume and reject the first wavelength; means for moving said excitation light and gel relative to one another to scan the gel; at least one spectral filter for receiving the collected emitted light and passing light at said different wavelength while rejecting light at said predetermined wavelength; a spatial filter for receiving the emitted light and passing light from said sample volume while rejecting background and scattered light; detecting means for receiving said collective fluorescently emitted light passed by said spectral and spatial filters and providing output signals, and means for receiving and processing said signals to provide an image of the labeled separated sample components.

Claim 7

A scanner for scanning a gel to detect fluorescence from fluorescence labeled separated sample components comprising means for supporting a gel to be scanned, means for applying excitation light at a predetermined wavelength to a volume in the gel to cause labeled, separated sample components to fluoresce at a different wavelength, and for collecting the fluorescence emitted light from said sample at said different wavelength from said volume and reject the first wavelength; means for moving said excitation light and gel relative to one another to scan the gel; at least one spectral filter for receiving the collected emitted light and passing light at said different wavelength while rejecting light at said predetermined wavelength; a spatial filter for receiving the emitted light and passing light from said sample volume while rejecting background and scattered light; detecting means for receiving said collective fluorescently emitted light passed by said spectral and spatial filters and providing output signals, and means for receiving and processing said signals to provide an image of the labeled separated sample components.

Claim 8

A scanner as in claim 4 including a laser for supplying said excitation light.

Claim 9

An improved scanner as in claim 5 including a spatial filter for receiving said light beam and transmitting light to said dichroic beam splitter.

Claim 10

An improved scanner for detecting fluorescence from a fluorescence labeled sample material carried by a sample carrier comprising means for forming a light beam having a predetermined wavelength; a dichroic beam splitter for receiving said light beam and directing it towards an objective lens which receives said beam and focuses the beam at a spot on the sample carrier whereby to cause the sample material to fluoresce at a selected volume and for collecting the fluorescently emitted light from said selected volume and directing the emitted light to said dichroic beam splitter, said fluorescent light being at a second wavelength and said dichroic beam splitter serving to pass the second wavelength; a spatial filter for receiving light at said second wavelength and passing light emitted from said selected volume; a spectral filter for further filtering said light and passing light at said second wavelength; means for moving the light beam and sample carrier relative to one another to scan the sample carrier, means for detecting the passed, emitted light of said second wavelength and providing output signals representative of the intensity of said detected light, and means for receiving and processing said signals to provide an image of the fluorescence of said samples.

Claim 11

An improved scanner for detecting fluorescence from a fluorescence labeled sample material carried by a sample carrier comprising means for forming a light beam having a predetermined wavelength; a dichroic beam splitter for receiving said light beam and directing it towards an objective lens which receives said beam and focuses the beam at a spot on the sample carrier whereby to cause the sample material to fluoresce at a selected volume and for collecting the fluorescently emitted light from said selected volume and directing the emitted light to said dichroic beam splitter, said fluorescent light being at a second wavelength and said dichroic beam splitter serving to pass the second wavelength; a spatial filter for receiving light at said second wavelength and passing light emitted from said selected volume; a spectral filter for further filtering said light and passing light at said second wavelength; means for moving the light beam and sample carrier relative to one another to scan the sample carrier, means for detecting the passed, emitted light of said second wavelength and providing output signals representative of the intensity of said detected light, and means for receiving and processing said signals to provide an image of the fluorescence of said samples.

Claim 12

An improved fluorescence scanner as in claim 7 in which the carrier is a gel and the sample is a nucleic acid or derivative thereof.

Claim 13

An improved gel scanner comprising means for supporting a gel to be scanned, means for forming a light beam of predetermined wavelength; a dichroic beam splitter for receiving and transmitting said beam toward a supported gel to be scanned; an objective lens for receiving said beam and focusing the beam in a selected volume of gel to cause fluorescence emission of light at a different wavelength and collecting the emitted light from samples in said selected volume and directing the emitted light to said dichroic beam splitter which reflects said emitted light of different wavelengths and rejects light at said predetermined wavelength; a spatial filter for receiving and passing the emitted light from said volume while rejecting background and scattered light; means for providing relative movement between said focused beam and the gel to be scanned, means for detecting said passed, emitted light and providing output signals, and means for processing said signals to provide an image of the fluorescence from the gel.

Claim 14

An improved gel scanner comprising means for supporting a gel to be scanned, means for forming a light beam of predetermined wavelength; a dichroic beam splitter for receiving and transmitting said beam toward a supported gel to be scanned; an objective lens for receiving said beam and focusing the beam in a selected volume of gel to cause fluorescence emission of light at a different wavelength and collecting the emitted light from samples in said selected volume and directing the emitted light to said dichroic beam splitter which reflects said emitted light of different wavelengths and rejects light at said predetermined wavelength; a spatial filter for receiving and passing the emitted light from said volume while rejecting background and scattered light; means for providing relative movement between said focused beam and the gel to be scanned, means for detecting said passed, emitted light and providing output signals, and means for processing said signals to provide an image of the fluorescence from the gel.