Fast-Indexing Filter Wheel and Method of Use

Abstract

Various embodiments of a sequencing system capable of rapidly imaging samples at multiple wavelengths are provided herein. In one embodiment, the system includes a fast-indexing filter wheel having a plurality of excitation and emission filters capable of being rapidly rotated into and out of communication with an excitation source (e.g., an arc lamp, a laser. For example, the filter wheel can be configured to index in an amount of time falling within a range of about 40 ms to about 60 ms, preferably 50 ms. The system can also be configured to account for vibrations resulting from the quick starts and stops of the fast-indexing filter wheel as well as vibrations resulting from other sources. Various methods of rapidly imaging a sample at multiple wavelengths are also provided herein.

1 .- 19 . (canceled) 
     
     
         20 . A method of analyzing a sample for nucleic acid sequencing, comprising:
 providing a fast-indexing filter wheel having a plurality of excitation filters;   positioning a first excitation filter of the plurality into optical communication with an excitation source;   irradiating an area of a sample with excitation energy such that a first emission signal is generated, the sample comprising a plurality of polynucleotides;   indexing a second excitation filter of the plurality into optical communication with the excitation source;   irradiating the area of the sample with a second excitation energy from the excitation source such that a second emission signal is generated;   detecting at least one of the emission signals from the irradiated area of the sample; and   moving the sample relative to the fast-moving filter wheel such that another area of the sample is positioned to be irradiated by at least the first excitation energy.   
     
     
         21 . The method of  claim 20 , comprising repeating the first irradiating step, the indexing step, and the second irradiating step for each of the plurality of filters prior to moving the sample. 
     
     
         22 . The method of  claim 21 , further comprising detecting each of the emission signals corresponding to each of the plurality of filters prior to moving the sample. 
     
     
         23 . The method of  claim 20 , wherein the fast-indexing filter wheel indexes between filters in an amount of time within a range of about 40 ms to about 60 ms. 
     
     
         24 . The method of  claim 23 , wherein the amount of time is within a range of about 45 ms to about 55 ms. 
     
     
         25 . The method of  claim 24 , wherein the amount of time is about 50 ms. 
     
     
         26 . The method of  claim 20 , wherein the fast-indexing filter wheel includes at least 4 excitation filters. 
     
     
         27 . The method of  claim 26 , indexing the at least 4 excitation filters prior to moving the sample. 
     
     
         28 . The method of  claim 20 , wherein the fast-indexing filter wheel includes 5 excitation filters. 
     
     
         29 . The method of  claim 28 , indexing the 5 excitation filters prior to moving the sample. 
     
     
         30 . The method of  claim 20 , wherein the fast-indexing filter wheel further includes at least one emission filter. 
     
     
         31 . The method of  claim 20 , wherein the fast-indexing filter wheel further includes a plurality of emission filters and a plurality of excitation filters. 
     
     
         32 . The method of  claim 31 , wherein each emission filter is positioned substantially perpendicular relative to one of the plurality of excitation filters. 
     
     
         33 . The method of  claim 20 , wherein the excitation source is an arc lamp. 
     
     
         34 . The method of  claim 20 , wherein the excitation source is a laser. 
     
     
         35 . The method of  claim 20 , wherein an anti-vibration mechanism is coupled to the filter wheel. 
     
     
         36 . The method of  claim 20 , wherein the sample is in a sample chamber or flowcell held by a processing stage. 
     
     
         37 . The method of  claim 36 , wherein a temperature block is in communication with the processing stage. 
     
     
         38 . The method of  claim 36 , wherein the processing stage moves relative indexing of the filter wheel as the another area of the sample is positioned to be irradiated. 
     
     
         39 . A method of analyzing a sample for nucleic acid sequencing, comprising:
 providing a fast-indexing filter wheel having a plurality of excitation filters;   positioning a first excitation filter of the plurality into optical communication with an excitation source;   irradiating an area of a sample with excitation energy such that a first emission signal is generated, the sample comprising a plurality of polynucleotides;   indexing a second excitation filter of the plurality into optical communication with the excitation source;   irradiating the area of the sample with a second excitation energy from the excitation source such that a second emission signal is generated;   detecting at least one of the emission signals from the irradiated area of the sample; and   moving the fast-moving filter wheel relative to the sample such that another area of the sample is positioned to be irradiated by at least the first excitation energy.