MedTech Initiative 2020

Genome Sequencing - Sequencing-by-Synthesis Method
Precision drives for fast and precise flow cell scanning in XYZ-directions andPiezo-based flow actuators for flow cells & other microfluidic cartridges

Physik Instrumente (PI) GmbH & Co. KG, Auf der Roemerstrasse 1, 76228 Karlsruhe, Germany
Phone +49 721 4846-0, Fax +49 721 4846-1019, Email info@pi.ws, www.pi.ws

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Genome Sequencing - Sequencing-by-Synthesis Method

Sequencing-by-Synthesis Method
The Sequencing-by-Synthesis method, also known as SolexaIllumina-method, is the most widespread method among all
Next-Generation-Sequencing methods (NGS). It was used toidentify the RNA fingerprint of the SARS-COV2 virus in January
2020 in a research institute in Wuhan. Since then, this RNA
fingerprint measurement has frequently been repeated inresearch institutes all over the world. And today this NGS
method is used worldwide day-by-day to continuously
monitor the SARS COV2 virus for mutations.
In addition to microfluidic flushing and washing cycles, as also
used by the other methods, e.g. the semiconductor-method
and the nanopore-method, the Sequencing-by-Synthesis
method is based on very precise and fast XYZ-optical image
scanning of the whole sample area.
To reduce turbulence and to achieve laminar flows at highspeed flushing and washing cycles, flow cells with several
parallel channels are used. Typical channel width is somemillimetres. This microfluidic pumping and sucking can bedone fast, precise and reliable by piezo-based actuators inform of membrane pump plates, benders or tubes.

fluorescence signal. The base pairs contain four different
nucleotides to result in four different fluorescence colors.

Recombined base pair read out by optical fluorescence signal

After each nucleotide flushing the read-out of the whole flow
cell has to be done by a XY-scan with all four laser excitation
wavelengths. For this purpose a fast and precise motion drive
combination for the laser excitation and read-out in every XYposition and at the correct Z-level is needed. Comprising aZ-focus adjustment in every XY-coordinate and then scanning
the XY-plane typically in a meander-like style.

Thymin fluorescence of a base pair at one strand position
Channels in a flow cell

The whole genome is put to many shorter pieces by the shotgun technique, these pieces are PCR amplified, then split tosingle strands (oligo strands) and finally attached to thebottom of the flow cell.

This results in a two-dimensional fluorescence data set after
each flushing & washing cycle. Each fluorescence signal
represents one nucleotide in a recombined base pair.

Resulted data set after one flushing & washing cycle
Oligo strands attached to the bottom of a flow cell channel

From here the stepwise recombination of base pairs starts
vertically and each recombination is indicated by aPhysik Instrumente (PI) GmbH & Co. KG, Auf der Roemerstrasse 1, 76228 Karlsruhe, Germany
Phone +49 721 4846-0, Fax +49 721 4846-1019, Email info@pi.ws, www.pi.ws

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Genome Sequencing - Sequencing-by-Synthesis Method

Motion Solutions from PI

Piezo actuators from PI

For the XYZ-scanning three drives are needed and typically inaddition a rotary drive to correctly position the whole flow
cell after cell exchange in the device. This means, in total four
precision drives are needed:

Piezo actuators operate at frequencies of up to several
kHz and are ideally suited for milli-, micro- andnanodispensing applications: They can switch valves
directly, work against a closing spring or a flexible tube forvolume displacement and propel liquids within amicrofluidic reaction volume – like a membrane pump.

Z-drive (focus drive). Typical requirements are: max.
stroke >5 mm; max. speed >25 mm/s; payload >500 g;
repeatability <300 nm; accuracy <3 µm
PICMA® Stack Multilayer Piezo Actuators

Voice coil drives
PICMA® Chip Actuators

XY-drives (sample stage). Typical requirements are: max.
travel range >100 mm x 100 mm; max. speed >200 mm/s;
payload >2 kg; repeatability <500 nm; accuracy <3 µm;
step & settle in raster scan 1,5 mm in < 40 ms.

PICMA® Multilayer Bender Actuators

Piezoelectric Components
3-phase magnetic direct drives, piezo motor stages

Rotary-drive (sample alignment after exchange). Typicalrequirements are: max. stroke >10°; max. speed >5°/s;
payload >0,5 kg; repeatability <0,1 mrad;
accuracy <1 mrad

DC motor drives, 3-phase magnetic direct drives

Miniaturized Piezo Components

Piezo Actuators and Components for Pumping Applications

Physik Instrumente (PI) GmbH & Co. KG, Auf der Roemerstrasse 1, 76228 Karlsruhe, Germany
Phone +49 721 4846-0, Fax +49 721 4846-1019, Email info@pi.ws, www.pi.ws

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ACS Motion Control

PI Subsidiaries

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FLY61E Genome Sequencing 04/2020 Subject to change without notice. © Physik Instrumente (PI) GmbH & Co. KG 2020

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