Flow cell systems and devices
Flow cell devices, cartridges, and systems are described that provide reduced manufacturing complexity, lowered consumable costs, and flexible system throughput for nucleic acid sequencing and other chemical or biological analysis applications. The flow cell device can include a capillary flow cell device or a microfluidic flow cell device.
1. A method of processing a sample of a subject having or suspected of having Coronavirus Disease 2019 (COVID-19) caused by a severe respiratory syndrome 2 (SARS-CoV-2) virus, said method comprising:
(a) providing a nucleic acid sequencing flow cell comprising two or more microfluidic channels, wherein at least one of said two or more microfluidic channels comprises a surface comprising a nucleic acid sequence immobilized thereto, wherein said nucleic acid sequence is derived from said sample of said subject having or suspected of having said COVID-19;
(b) contacting said nucleic acid sequence with a primer having complementarity with said nucleic acid sequence, to yield a primed nucleic acid sequence;
(c) using at least (i) a polymerizing enzyme and (ii) a plurality of nucleobases having a plurality of detectable labels to subject said primed nucleic acid sequence to a primer extension reaction;
(d) while performing said primer extension reaction, using a detector to detect a plurality of signals from at least a subset of said plurality of detectable labels; and
(e) using said plurality of signals detected in (d) to generate sequencing data;
(f) using one or more logic circuits to process said sequencing data in parallel to identify said nucleic acid sequence; and
(g) using said nucleic acid sequence identified in (f) to determine that said sample contains a messenger ribonucleic acid (mRNA) molecule encoding said SARS-CoV-2.
2. The method of claim 1 , wherein said plurality of detectable labels comprises a plurality of fluorophores.
3. The method of claim 1 , wherein said using said plurality of signals detected in (d) to identify said nucleic acid sequence comprises imaging a first surface and an axially-displaced second surface of said two or more microfluidic channels using a fluorescent imaging system.
4. The method of claim 1 , further comprising, before (a): (h) introducing a fluid composition comprising said nucleic acid sequence at a concentration of between about 0.5 nanomolar (nM) and 900 nM to said nucleic acid sequencing flow cell.
5. The method of claim 1 , further comprising, before (a):
(i) hybridizing at least a portion of said nucleic acid sequence to at least a portion of oligonucleotides grafted to said surface.
6. The method of claim 4 , wherein said nucleic acid sequencing flow cell is loaded with said fluid composition prior to (c).
7. The method of claim 1 , wherein said nucleic acid sequence is present at said surface at a surface density comprising at least 4,000 molecules per square micrometer (μm 2 ).
8. The method of claim 5 , wherein said surface is silanized and said oligonucleotides are covalently grafted to said surface.
9. The method of claim 1 , wherein at least one of said two or more microfluidic channels is configured to hold a fluid volume between about 10 to about 300 microliters.
10. The method of claim 1 , wherein a fluorescent image of said surface exhibits a contrast-to-noise ratio of at least or equal to about 5 when said fluorescent image of said surface is obtained with an inverted fluorescence microscope and a camera under non-signal saturating conditions while said surface is immersed in a buffer, and said plurality of detectable labels comprise Cyanine dye-3.
11. The method of claim 1 , wherein said nucleic acid sequence has a length comprising more than or equal to about 100 base pairs.
12. The method of claim 1 , wherein said nucleic acid sequencing flow cell comprises a flow cell cartridge that mates with a microfluidic chip.
13. The method of claim 1 , further comprising, before (a):
(j) extracting said mRNA from said sample; and
(k) converting said mRNA to complementary DNA comprising said nucleic acid sequence.
14. The method of claim 1 , wherein said sample comprises epithelial cells.
15. The method of claim 1 , wherein said one or more logic circuits comprises a field programmable gate array (FPGA).
16. The method of claim 1 , wherein said sequencing data is further analyzed using a central processing unit (CPU).
17. The method of claim 1 , wherein using said one or more logic circuits to process said sequencing data in parallel is performed with Cloud-based computing.
18. The method of claim 1 , wherein said two or more microfluidic channels comprises four microfluidic channels.
19. The method of claim 1 , further comprising amplifying said nucleic acid sequence prior to (a) or (b) using polymerase chain reaction (PCR).
20. The method of claim 1 , wherein said performing said primer extension reaction comprises performing a sequencing-by-synthesis reaction.