Ruggedized apparatus for analysis of nucleic acid and proteins

Abstract

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.

What is claimed is: 
     
         1 - 26 . (canceled) 
     
     
         27 . A system for finding the position of at least one or a plurality of separation channels in a removable electrophoresis chip, said system comprising:
 (a) said removable electrophoresis chip comprising at least one or a plurality of channels, wherein said channel or each of said plurality of channels comprises a detection zone formed of a material that allows at least a part of a fluorescence excitation energy beam to interact with said channel or plurality of separation channels; and   (b) an electrophoresis instrument capable of receiving said removable electrophoresis chip, and configured to separate and detect at least one fluorescently labeled nucleic acid fragment, comprising,
 (i) a data acquisition and storage system comprising a computer; 
 (ii) a fluorescence excitation and detection system further comprising a component for detecting fluorescence from said at least one fluorescently labeled nucleic acid fragment; and, 
 (iii) a lanefinding system comprising an excitation energy beam source configured to produce fluorescence, and at least one detector positioned and configured to detect fluorescence intensities from said material, and from each of said one or a plurality of detection zones, and for a plurality of channels, between each one of said plurality of channels, and to send said fluorescence intensity data, to said data acquisition and storage system, 
 and whereby in use the computer resolves the differences in fluorescence intensities to determine the location of each one or plurality of separation channels. 
   
     
     
         28 . The system  claim 27  wherein the computer further comprises baseline subtraction to determine and remove the background signal component comprising all detectable components except for the fluorescent signal from said at least one nucleic acid fragment. 
     
     
         29 . The system of  claim 27  wherein the light detector comprises at least one photomultiplier tube. 
     
     
         30 . The system of  claim 27  wherein the light detector is a multi-element PMT. 
     
     
         31 . The system of  claim 27  wherein the light detector is a CCD camera. 
     
     
         32 . The system of  claim 27  wherein the light detector is a linear array detector. 
     
     
         33 . The system of  claim 27  wherein the one or plurality of separation channels are formed from glass. 
     
     
         34 . The system of  claim 27  wherein the one or plurality of separation channels are formed from polymer or co-polymer. 
     
     
         35 . An apparatus for processing a sample of biomolecular analyte, the apparatus comprising:
 a. a test module comprising a material that allows at least a part of an energy source 1) to interact with the sample disposed within the test module and 2) to interact with at least one channel disposed within the test module, with the area near said channel, and for a plurality of separation channels, in the area between each one of said plurality of channels;   b. said at least one channel in fluid communication with the sample;   c. a holder for supporting the test module;   d. an electrophoresis device connected to the holder and for providing energy to the test module;   e. a light source for emitting a light beam that excites fluorescence in the sample of biomolecular analyte and which excites fluorescence in the channel and the area near said channel;   f. at least one light detector positioned and configured to detect differences in fluorescence intensities from said material, and from and within each one of said plurality of channels, and from between said plurality of channels, and to send fluorescence intensity signals;   g. a computer for receiving fluorescence intensity signals from said at least one light detector and for determining a location of the at least one channel disposed within the test module.   
     
     
         36 . The apparatus of  claim 35 , wherein the holder for supporting the test module comprises: a first member including a pair of electrodes disposed to electrically connect with openings disposed on a test module; and a second member including an automated locking feature to reduce lateral motion of the test module when the test module is positioned between the first member and the second member. 
     
     
         37 . The apparatus of  claim 35 , wherein the at least one channel comprises at least one microfluidic channel or capillary. 
     
     
         38 . The apparatus of  claim 35 , wherein the test module is glass. 
     
     
         39 . The apparatus of  claim 35 , wherein the test module is polymethyl methacrylate, UV-treated polycarbonate or cyclic olefin copolymer. 
     
     
         40 . The apparatus of  claim 35 , wherein the test module is single crystal alumina. 
     
     
         41 . The apparatus of  claim 35 , wherein the test module is a clear polymer or copolymer. 
     
     
         42 . The apparatus of  claim 35 , wherein the light detector is a photodiode. 
     
     
         43 . The apparatus of  claim 35 , wherein the light detector is at least one photomultiplier tube. 
     
     
         44 . The system of  claim 35  wherein the light detector is a multi-element PMT. 
     
     
         45 . The apparatus of  claim 35  wherein the light detector is a CCD camera. 
     
     
         46 . The system of  claim 35  wherein the light detector is a linear array detector. 
     
     
         47 . The apparatus of  claim 35  wherein the computer further comprises baseline subtraction to determine and remove the background signal component comprising all detectable components except for the fluorescent signal from said at least one biomolecular analyte. 
     
     
         48 . A system for separation and detection of nucleic acid fragments comprising:
 a. a removeable electrophoresis capillary or chip formed of a material that allows at least a part of a fluorescence excitation energy beam to interact with said material, comprising: one or a plurality of channels, each one or plurality of channels having a detection zone;   b. an electrophoresis instrument comprising an optical and electrical coupling to said chip, and
 (i) a data acquisition and storage system comprising a computer; 
 (ii) a fluorescence excitation and detection system for detecting fluorescence from labeled nucleic acid fragments; and 
 (iii) a lanefinding system comprising an excitation energy beam source configured to produce fluorescence; at least one detector positioned and configured to detect fluorescence from said material, and within or between each of said one or a plurality of channels, and a data input to said data acquisition and storage system; 
   
       whereby in use the detector detects fluorescence intensity signals within each of said one or plurality of channels, from said material comprising said capillary or chip, and, for a plurality of channels, from the area between each of said plurality of channels, and the computer resolves the differences in fluorescence intensity signals to determine the location of each channel. 
     
     
         49 . The system of  claim 48  wherein the light detector is at least one PMT. 
     
     
         50 . The system of  claim 48  wherein the light detector is a multi-element PMT. 
     
     
         51 . The apparatus of  claim 48  wherein the light detector is a CCD camera. 
     
     
         52 . The system of  claim 48  wherein the light detector is a linear array detector.