01 The bigger picture
Biology doesn't happen one target at a time. Your analysis shouldn't have to, either.
Meet Hyperplex PCR™. The familiarity of PCR, expanded to measure many targets in a single well. Look wider, without splitting your sample into a separate reaction for every question.
02 Inside Hyperplex PCR™
From sequence to signal.
Four steps.
A different way to see your targets.
Amplify
Start from extracted DNA or RNA and amplify selected target sequences in high multiplex PCR.
DNA
Selected target sequences
Many targets. Amplified together.
Multiplex PCR in one tube
Symbolic duplex target sequences from several target families before and after amplification. Both views represent the same reaction tube, not a transfer between tubes. The number of symbols is illustrative.
Selected targets
Amplified targets
One tube · multiplex PCR
Recognise
Padlock probes bind to target sequences and form circular templates through sequence-specific ligation, providing single-nucleotide specificity at the junction.
PLP
Padlock probe · bound to its target
Ligation-driven specificity.
Padlock recognition at the ligation junction
Two padlock probes each contain a centred embedded barcode. Four equally spaced base positions pair with each target. Matching terminal bases permit ligation; a single-base mismatch adjacent to the junction disfavors ligation and the probe is shown open. Geometry is schematic.
Embedded barcode
Match
Ligation
Mismatch
Disfavours ligation
Sequence recognition at the junction
Build a countable signal
Rolling-circle amplification converts circularised probes into individual RCA products, carrying repeated copies of their target-specific barcode.
RCP
Rolling circle product
One product. Repeated target-specific barcodes.
Rolling-circle amplification in one tube
Closed circular templates bearing embedded barcode sequences are amplified into separate long coiled RCA products. Each target keeps its pink, cyan or green family, with a lighter backbone and saturated barcode segments. All templates have equal diameter; all products use the same loose-coil shape and size. The circle is a template, not a strand unrolling into the product. Both views represent one reaction tube before and after RCA.
Circular templates
RCA products
One tube · rolling-circle amplification
Nanopixel labelling
A universal, single-tube mix combines hundreds of Nanopixel codes. Complementary DNA probes bind Nanopixels to target-specific RCP barcodes. Their fluorescence colour and intensity identify each target.
Nanopixel
Fluorescent barcode label
Target identity, made visible.
Nanopixel labelling
Three target-specific RCPs on a plate-well surface after unbound Nanopixels have been washed away. The cyan RCP is enlarged. Each loose coil carries multiple matching Nanopixels, with rounded detection oligos and four paired bases per binding site.
Nanopixels
Barcode
Unbound Nanopixels are washed away
Optical encoding
Illustrative codes · three channels, four displayed levels. Not a calibrated assay codebook.
- Code 01: illustrative relative contributions 1, 4, 4.
- Code 02: illustrative relative contributions 4, 1, 3.
- Code 03: illustrative relative contributions 1, 4, 1.
03 The optical readout
From images to counts.
More of the well imaged. More counts collected.
Our software analyses images as they are acquired, resolving up to 200 Nanopixel codes into a count matrix.
- Standard 1M+ counts
- DeepCount 30M+ counts
Up to 200 Nanopixel codes.
100 Nanopixel code example
100 illustrative Nanopixel codes in five colour groups.
Click a cluster to isolate its RCPs.
| Target | Nanopixel code | Standard · Well A01 | DeepCount · Well A01 |
|---|---|---|---|
| Target 01 | 01 | 11,953 | 358,594 |
| Target 02 | 02 | 10,234 | 307,031 |
| Target 03 | 03 | 9,141 | 274,219 |
| Target 04 | 04 | 9,688 | 290,625 |
| Target 05 | 05 | 10,313 | 309,375 |
| Target 06 | 06 | 9,531 | 285,938 |
| Target 07 | 07 | 8,828 | 264,844 |
| Target 08 | 08 | 10,000 | 300,000 |
| Target 09 | 09 | 11,172 | 335,156 |
| Target 10 | 10 | 8,828 | 264,844 |
| Target 11 | 11 | 9,375 | 281,250 |
| Target 12 | 12 | 9,922 | 297,656 |
| Target 13 | 13 | 11,328 | 339,844 |
| Target 14 | 14 | 9,688 | 290,625 |
| Target 15 | 15 | 9,688 | 290,625 |
| Target 16 | 16 | 10,234 | 307,031 |
| Target 17 | 17 | 10,625 | 318,750 |
| Target 18 | 18 | 9,297 | 278,906 |
| Target 19 | 19 | 10,156 | 304,688 |
| Target 20 | 20 | 11,094 | 332,813 |
| Target 21 | 21 | 9,766 | 292,969 |
| Target 22 | 22 | 11,953 | 358,594 |
| Target 23 | 23 | 11,953 | 358,594 |
| Target 24 | 24 | 10,938 | 328,125 |
| Target 25 | 25 | 8,594 | 257,813 |
| Target 26 | 26 | 8,359 | 250,781 |
| Target 27 | 27 | 8,516 | 255,469 |
| Target 28 | 28 | 9,766 | 292,969 |
| Target 29 | 29 | 10,078 | 302,344 |
| Target 30 | 30 | 10,703 | 321,094 |
| Target 31 | 31 | 11,172 | 335,156 |
| Target 32 | 32 | 9,688 | 290,625 |
| Target 33 | 33 | 9,766 | 292,969 |
| Target 34 | 34 | 10,469 | 314,063 |
| Target 35 | 35 | 10,234 | 307,031 |
| Target 36 | 36 | 8,438 | 253,125 |
| Target 37 | 37 | 9,375 | 281,250 |
| Target 38 | 38 | 8,750 | 262,500 |
| Target 39 | 39 | 10,313 | 309,375 |
| Target 40 | 40 | 10,859 | 325,781 |
| Target 41 | 41 | 8,359 | 250,781 |
| Target 42 | 42 | 10,938 | 328,125 |
| Target 43 | 43 | 8,984 | 269,531 |
| Target 44 | 44 | 9,219 | 276,563 |
| Target 45 | 45 | 10,859 | 325,781 |
| Target 46 | 46 | 9,062 | 271,875 |
| Target 47 | 47 | 9,531 | 285,938 |
| Target 48 | 48 | 11,641 | 349,219 |
| Target 49 | 49 | 10,859 | 325,781 |
| Target 50 | 50 | 11,797 | 353,906 |
| Target 51 | 51 | 8,828 | 264,844 |
| Target 52 | 52 | 9,219 | 276,563 |
| Target 53 | 53 | 12,031 | 360,938 |
| Target 54 | 54 | 10,312 | 309,375 |
| Target 55 | 55 | 10,547 | 316,406 |
| Target 56 | 56 | 9,844 | 295,313 |
| Target 57 | 57 | 11,172 | 335,156 |
| Target 58 | 58 | 9,375 | 281,250 |
| Target 59 | 59 | 10,547 | 316,406 |
| Target 60 | 60 | 10,469 | 314,063 |
| Target 61 | 61 | 9,609 | 288,281 |
| Target 62 | 62 | 9,375 | 281,250 |
| Target 63 | 63 | 10,547 | 316,406 |
| Target 64 | 64 | 10,000 | 300,000 |
| Target 65 | 65 | 10,391 | 311,719 |
| Target 66 | 66 | 10,156 | 304,687 |
| Target 67 | 67 | 9,687 | 290,625 |
| Target 68 | 68 | 9,062 | 271,875 |
| Target 69 | 69 | 9,531 | 285,937 |
| Target 70 | 70 | 10,703 | 321,094 |
| Target 71 | 71 | 11,641 | 349,219 |
| Target 72 | 72 | 8,437 | 253,125 |
| Target 73 | 73 | 10,547 | 316,406 |
| Target 74 | 74 | 9,922 | 297,656 |
| Target 75 | 75 | 9,609 | 288,281 |
| Target 76 | 76 | 10,156 | 304,687 |
| Target 77 | 77 | 10,156 | 304,687 |
| Target 78 | 78 | 11,172 | 335,156 |
| Target 79 | 79 | 9,766 | 292,969 |
| Target 80 | 80 | 10,859 | 325,781 |
| Target 81 | 81 | 10,781 | 323,437 |
| Target 82 | 82 | 8,828 | 264,844 |
| Target 83 | 83 | 8,984 | 269,531 |
| Target 84 | 84 | 9,375 | 281,250 |
| Target 85 | 85 | 9,844 | 295,312 |
| Target 86 | 86 | 11,641 | 349,219 |
| Target 87 | 87 | 9,922 | 297,656 |
| Target 88 | 88 | 9,531 | 285,937 |
| Target 89 | 89 | 9,844 | 295,312 |
| Target 90 | 90 | 9,922 | 297,656 |
| Target 91 | 91 | 9,766 | 292,969 |
| Target 92 | 92 | 11,016 | 330,469 |
| Target 93 | 93 | 8,437 | 253,125 |
| Target 94 | 94 | 10,859 | 325,781 |
| Target 95 | 95 | 8,437 | 253,125 |
| Target 96 | 96 | 10,312 | 309,375 |
| Target 97 | 97 | 9,687 | 290,625 |
| Target 98 | 98 | 8,125 | 243,750 |
| Target 99 | 99 | 9,219 | 276,562 |
| Target 100 | 100 | 9,766 | 292,969 |
| All 100 targets | 1,000,000 | 30,000,000 |