Instrument for monitoring polymerase chain reaction of dna

Abstract

An optical instrument monitors PCR replication of DNA in a reaction apparatus having a temperature cycled block with vials of reaction ingredients including dye that fluoresces in presence of double-stranded DNA. A beam splitter passes an excitation beam to the vials to fluoresce the dye. An emission beam from the dye is passed by the beam splitter to a CCD detector from which a processor computes DNA concentration. A reference strip with a plurality of reference emitters emit reference beams of different intensity, from which the processor selects an optimum emitter for compensating for drift. Exposure time is automatically adjusted for keeping within optimum dynamic ranges of the CCD and processor. A module of the beam splitter and associated optical filters is associated with selected dye, and is replaceable for different dyes.

1 - 17 . (canceled) 
     
     
         18 . An instrument comprising:
 an apparatus configured to hold a plurality of spaced-apart reaction regions;   a light source configured to emit an excitation beam;   a detector configured to receive emission beams from the reaction regions;   a first path from the light source to the apparatus;   a second path from the apparatus to the detector; and   an optical system comprising a lens, the lens disposed along the first path and along the second path, the optical system configured to direct the excitation beam to more than one of the reaction regions simultaneously.   
     
     
         19 . The instrument of  claim 18 , wherein the detector is configured to generate data signals representative of the emission beams. 
     
     
         20 . The instrument of  claim 18 , wherein the light source comprises a laser. 
     
     
         21 . The instrument of  claim 18 , further comprising a dichroic reflector configured to reflect the excitation beam and to transmit the emissions to the detector. 
     
     
         22 . The instrument of  claim 18 , wherein the light source emits at least two excitation beams, each excitation beam characterized by a central wavelength that is different from that of the other excitation beam(s). 
     
     
         23 . The instrument of  claim 23 , further comprising a plurality of dichroic reflectors configured to reflect the at least two excitation beams from the light source. 
     
     
         24 . The instrument of  claim 18 , wherein lens comprises a focusing optic. 
     
     
         25 . An instrument comprising:
 an apparatus comprising a portion configured to hold a plurality of spaced-apart reaction regions;   a light source configured to emit an excitation beam;   a detector configured to receive emission beams from the reaction regions; and   an optical system comprising a lens, the optical system configured to direct the excitation beam through the lens to more than one of the reaction regions simultaneously;   a first path from the lens to the portion, the instrument configured to direct the excitation beam along the first path from the light source to the portion;   a second path from the portion and through the lens, the instrument configured to direct emissions along the second path from the plurality of spaced-apart reaction regions to the detector.   
     
     
         26 . The instrument of  claim 25 , wherein the detector is configured to generate data signals representative of the emission beams. 
     
     
         27 . The instrument of  claim 25 , wherein the light source comprises a laser. 
     
     
         28 . The instrument of  claim 25 , further comprising a dichroic reflector configured to reflect the excitation beam and to transmit the emissions to the detector. 
     
     
         29 . The instrument of  claim 25 , wherein the light source emits at least two excitation beams, each excitation beam characterized by a central wavelength that is different from that of the other excitation beam(s). 
     
     
         30 . The instrument of  claim 29 , further comprising a plurality of dichroic reflectors configured to reflect light from the light source. 
     
     
         31 . The instrument of  claim 25 , wherein lens comprises a focusing optic. 
     
     
         32 . An instrument comprising:
 an apparatus comprising a surface configured to hold a plurality of spaced-apart reaction regions;   a light source configured to emit an excitation beam;   a detector configured to receive emission beams from the reaction regions; and   an optical system comprising a lens, the optical system configured to direct the excitation beam through the lens to more than one of the reaction regions simultaneously;   wherein the optical system is configured to direct emission beams through the lens to the detector.   
     
     
         33 . The instrument of  claim 32 , wherein the detector is configured to generate data signals representative of the emission beams. 
     
     
         34 . The instrument of  claim 32 , wherein the light source comprises a laser. 
     
     
         35 . The instrument of  claim 32 , further comprising a dichroic reflector configured to reflect the excitation beam and to transmit the emissions to the detector. 
     
     
         36 . The instrument of  claim 32 , wherein the light source emits at least two excitation beams, each excitation beam characterized by a central wavelength that is different from that of the other excitation beam(s). 
     
     
         37 . The instrument of  claim 36 , further comprising a plurality of dichroic reflectors configured to reflect light from the light source. 
     
     
         38 . The instrument of  claim 32 , wherein lens comprises a focusing optic.