US2009095919A1PendingUtilityA1

System and method for monitoring cellular activity

58
Individually held — no corporate assignee on recordPriority: Jul 30, 1999Filed: Dec 15, 2008Published: Apr 16, 2009
Est. expiryJul 30, 2019(expired)· nominal 20-yr term from priority
Inventors:Gregory BearmanScott E. FraserRussell D. Lansford
G01N 21/6458G01N 21/6428G01N 2021/6417Y10S435/968G01N 2021/6421
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Claims

Abstract

A system and method for monitoring cellular activity in a cellular specimen. According to one embodiment, a plurality of excitable markers are applied to the specimen. A multi-photon laser microscope is provided to excite a region of the specimen and cause fluorescence to be radiated from the region. The radiating fluorescence is processed by a spectral analyzer to separate the fluorescence into respective wavelength bands. The respective bands of fluorescence are then collected by an array of detectors, with each detector receiving a corresponding one of the wavelength bands.

Claims

exact text as granted — not AI-modified
1 . A system, comprising:
 a single photon laser configured to emit radiation capable of exciting a plurality of different excitable markers in a region of a cellular specimen to cause the plurality of different excitable markers to radiate florescence;   a focusing lens configured to focus radiation emitted by the single photon laser on the region of the cellular specimen;   a dichroic mirror configured to reflect the fluorescence;   a grating configured to separate the reflected fluorescence into wavelength bands of fluorescence;   a linear array of high gain photomultiplier tubes configured so that each of the wavelength bands of fluorescence is incident upon a corresponding one of the high gain photomultiplier tubes and so that each of the high gain photomultiplier tubes generates a corresponding analog signal;   a plurality of digital to analog converters, each digital to analog converter being configured to convert one of the analog signals from the linear array of high gain photomultiplier tubes to a digital signal; and   a processor configured to linearly unmix the digital signals.   
   
   
       2 . The system of  claim 1 , wherein the dichroic mirror is configured so that radiation from the single photon laser that is reflected from the specimen passes through the dichroic mirror. 
   
   
       3 . The system of  claim 1 , wherein the processor is configured to identify the contribution to the fluorescence from each of the plurality of excitable markers. 
   
   
       4 . A system, comprising:
 a single photon laser microscope operative to cause a plurality of different excitable markers in a region of a cellular specimen to radiate fluorescence;   photomultiplier tubes, each of the plurality of photomultiplier tubes being configured to receive a corresponding wavelength band of the florescence so that each of the photomultiplier tubes generates a corresponding analog signal;   a plurality of digital to analog converters, each digital to analog converter being configured to convert one of the analog signals from the linear array of high gain photomultiplier tubes to a digital signal; and   a processor configured to unmix the digital signals.   
   
   
       5 . The system of  claim 4 , wherein the single photon laser microscope comprises a single photon laser configured to emit the radiation. 
   
   
       6 . The system of  claim 5 , further comprising a focusing lens configured to focus the radiation to the region of the cellular specimen. 
   
   
       7 . The system of  claim 4 , further comprising a dichroic mirror configured to reflect the fluorescence. 
   
   
       8 . The system of  claim 7 , wherein the dichroic mirror is configured so that radiation from the single photon laser that is reflected from the specimen passes through the dichroic mirror. 
   
   
       9 . The system of  claim 4 , further comprising a grating configured to separate the fluorescence into wavelength bands of fluorescence. 
   
   
       10 . The system of  claim 4 , wherein the photomultiplier tubes form a linear array. 
   
   
       11 . The system of  claim 10 , wherein the photomultiplier tubes are high gain photomultiplier tubes. 
   
   
       12 . The system of  claim 4 , wherein the photomultiplier tubes are high gain photomultiplier tubes. 
   
   
       13 . The system of  claim 4 , wherein the processor is configured to linearly unmix the digital signals. 
   
   
       14 . The system of  claim 4 , wherein the processor is configured to identify the contribution to the fluorescence from each of the plurality of excitable markers. 
   
   
       15 . A method, comprising:
 using the system of  claim 1  to monitor cellular activity in a cellular specimen.   
   
   
       16 . A method, comprising:
 using the system of  claim 3  to monitor cellular activity in a cellular specimen.   
   
   
       17 . A method, comprising:
 applying a plurality of different excitable markers to a cellular specimen;   applying light to the cellular specimen from a single-photon laser microscope to cause fluorescence to be radiated from the region by markers in that region of the cellular specimen;   using a grating to separate the fluorescence into wavelength bands of fluorescence; and   using a linear array of photomultiplier tubes to detect the florescence, each photomultiplier tube receiving one of the wavelength bands of fluorescence and generating a corresponding analog signal;   converting the analog signals to digital signals; and   linearly unmixing the digital signals.   
   
   
       18 . The method of  claim 17 , further comprising identifying the contribution to the fluorescence from each of the plurality of excitable markers. 
   
   
       19 . A method, comprising:
 applying light from a single-photon laser to a region of a cellular specimen having a plurality of different excitable markers, thereby causing florescence to be radiated from the region of the cellular specimen;   separating the fluorescence into wavelength bands of florescence using a grating; and   detecting the fluorescence via photomultiplier tubes, each photomultiplier tube receiving one of the wavelength bands of florescence and generating a corresponding analog signal;   converting the analog signals to digital signals; and   linearly unmixing the digital signals.   
   
   
       20 . The method of  claim 19 , further comprising applying the plurality of different excitable markers to the cellular specimen. 
   
   
       21 . The method of  claim 19 , wherein the single photon laser is part of a single photon laser microscope. 
   
   
       22 . The method of  claim 19 , wherein the photomultiplier tubes form a linear array. 
   
   
       23 . The method of  claim 22 , wherein the photomultiplier tubes are high gain photomultiplier tubes. 
   
   
       24 . The method of  claim 19 , wherein the photomultiplier tubes are high gain photomultiplier tubes. 
   
   
       25 . The method of  claim 19 , further comprising identifying the contribution to the fluorescence from each of the plurality of excitable markers. 
   
   
       26 . A system, comprising:
 a single photon laser configured to emit radiation capable of exciting a plurality of different excitable markers in a region of a cellular specimen to cause the plurality of different excitable markers to radiate fluorescence;   a focusing lens configured to focus radiation emitted by the single photon laser on the region of the cellular specimen;   a dichroic mirror configured to reflect the fluorescence;   a grating configured to separate the reflected fluorescence into wavelength bands of fluorescence;   a linear array of high gain photomultiplier tubes configured so that each of the wavelength bands of fluorescence is incident upon a corresponding one of the high gain photomultiplier tubes and so that each of the high gain photomultiplier tubes generates a corresponding analog signal;   a plurality of digital to analog converters, each digital to analog converter being configured to convert one of the analog signals from the linear array of high gain photomultiplier tubes to a digital signal; and   a processor configured to identify the contribution to the fluorescence from each of the plurality of excitable markers.   
   
   
       27 . A system, comprising:
 a single photon laser microscope operative to cause a plurality of different excitable markers in a region of a cellular specimen to radiate fluorescence;   photomultiplier tubes, each of the plurality of photomultiplier tubes being configured to receive a corresponding wavelength band of the fluorescence so that each of the photomultiplier tubes generates a corresponding analog signal;   a plurality of digital to analog converters, each digital to analog converter being configured to convert one of the analog signals from the linear array of high gain photomultiplier tubes to a digital signal; and   a processor configured to identify the contribution to the fluorescence from each of the plurality of excitable markers.

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