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Frequently Asked Questions Regarding:Acoustic Droplet Ejection (ADE) TechnologyThe Echo® Series 500 Liquid Handling PlatformsThe Portrait® 630 SpotterDeerac™ Liquid Handling PlatformsAcoustic Droplet Ejection (ADE) Technology
What is ADE?
Acoustic droplet ejection (ADE) uses the energy of a sound pulse to propel a precise and accurate droplet of liquid from a source reservoir onto a surface suspended above the source. This suspended surface can be a multi-well plate, a piece of glass or a tissue sample. The
Echo® Liquid Handling Systems are designed to transfer liquids into microplates. The Portrait 630 is designed to transfer liquid onto tissue samples on MALDI targets. See
Technology for additional technical detail. Nothing comes in physical contact with the fluid. The system uses no pipette tips, pin tools or nozzles. There is no chance of cross-contamination of samples, and the requirement for wash procedures is eliminated. ADE is a gentle process that can transfer proteins, nucleic acids, or live cells without damage or loss of viability.
Labcyte has applied ADE to move drug compounds between multi-well microplates in our Echo liquid handler product line, and to deposit reagents onto tissue samples for MALDI tissue imaging in our
Portrait® 630 Reagent Multi-Spotter line. ADE technology can also be used form any other applications.
Back to main list.What volumes of fluid can be transferred?
ADE can eject droplets as small as one picoliter (10-12 liter) and as large as 10 microliters. The size of the droplet depends on several factors. The frequency of sound emitted by the acoustic transducer determines the size of the ejected droplet. Higher frequencies produce smaller droplets. The viscosity and surface tension of the liquid being ejected also influence the droplet volume. The droplet size is fixed in both the Echo and Portrait systems. Larger volumes are transferred by increasing the number of droplets ejected.
The Echo® 500 series liquid handlers eject discrete droplets of your solution. The minimum droplet volume for DMSO is 2.5 nL. The maximum volume for a single transfer is 1000 nL or four hundred droplets, which ensures that the transducer remains focused on the fluid meniscus. Larger volumes can be transferred with multiple transfer steps
Back to main list.How precise and accurate are the transfers?
The Echo systems are warranted to have no more than 10% deviation from the target volume and to have precision better than 8% CV over the entire volume transfer range of 2.5 to 250 nL. Users typically report values significantly better than the warranted numbers.
Back to main list.Does ADE technology sonicate materials?
Labcyte ADE technology introduces very small amounts of energy that is not enough to dissolve a solid or to break up a mass of particles. This is useful, for example, in experiments where ADE has been used to transfer cells. The cells did not suffer any adverse effects from the sonic energy.
Back to main list.Do samples become heated by the sonic energy?
No detectable increase in temperature is measured. The energy used is comparable to that used in medical diagnostic ultrasound.
Back to main list.Why do the instruments deionize the plates and reservoirs?
Plastic plates pick up static electricity. While the charge is not enough to cause a shock when you touch the plates, it can be enough to deflect the flight of nanoliter volume droplets as they fly from the source to the destination.
To eliminate this problem and ensure that the droplets fly true, Echo systems move both source and destination plates past deionizing bars that remove any residual charge. In addition, all Echo
® qualified plates from Labcyte are manufactured and packaged to reduce their inherent static charge. Plates from other vendors will not be pre-treated. Deionization during loading may not be sufficient to ensure proper transfer and placement of droplets during ejection.
In the Portrait system, the MALDI matrix liquid is deionized as it is moved into the system past the deionizing bars. The customer has the option of deionizing the tissue sample as it enters the system.
Back to main list.What is acoustic grade COC plastic? Why does Labcyte use it? How does it interact with DMSO? Is it good for long-term storage? Can COC plates be heat-sealed?
COC is cyclic olefin copolymer. This plastic is resistant to a wide range of biochemical reagents and solvents. In tests done to date, COC is comparable to polypropylene for heat-sealing and long-term storage of DMSO solutions. COC is much less permeable to water than polypropylene.
Labcyte prefers COC over polypropylene for ADE because sound travels through COC with less interference than when traveling through polypropylene. This improves the precision of hydration determination, fluid height evaluation and liquid transfer.
Back to main list.The Echo Series 500 Liquid Handlers
What are the Echo Series 500 liquid handlers?
The
Echo® Liquid Handling Systems are designed to use ADE to transfer nanoliter volumes of DMSO and aqueous solutions from source plates to destination plates. In both systems, liquid from any well in a source plate can be delivered to any well in a destination plate. The volume of the transfer can be set for each individual well or for the entire plate. Fluid from a single well in a source plate can be transferred to multiple wells of the destination plate in multiple volumes. Fluids from multiple source wells can be pooled into the same destination well if desired. GUI flexibility lets the user decide whether reformatting to a higher density plate should be done in quadrant mode or in interleaved mode.
The systems measure the fluid height in each source well and the percent hydration of DMSO-based samples. They also automatically confirm transfer success in each well. The Echo 555 liquid handler is approximately twice as fast as the Echo 550. See
“How fast do the Echo systems transfer plates?” for more detail.
Back to main list.How fast do the Echo systems transfer plates?
Cycle time is the time required to replicate a single plate. The Echo 555 has a cycle time of about half of that of the Echo 550 (see the charts below.) However the time to replicate a single plate is not as important as the time to deliver a complete set of assay-ready wells. Fixed head replicators often require both tip washing and intermediate plate set-up that add time to the process. These steps are completely eliminated with ADE, resulting in faster assay-ready throughput. Some examples follow.
Note that the analyses shown do not include reading barcodes, which adds approximately 1.5 seconds to the cycle time. In single-plate-to-single-plate replications, this would add an extra 3 seconds per cycle.
Large differences in sample depth (volume) can also add to the cycle time because the transducer will spend significant time adjusting the Z-axis focal position. (Most plates have roughly equal volume per well, so this is rarely an issue.) Gradients in volume over the plate, or a having a few cherry-picked wells with lower volume contribute little effect on throughput.
The analysis also assumes that the DMSO in each well has approximately the same amount of hydration, although hydration effects on throughput are much smaller than volume effects.
Finally, the throughput numbers require a robotic transfer time to be incorporated into the set-up to account for the robotic loading and unloading of plates. In the analysis shown, the robotic plate handling time is assumed to be 3 seconds per plate. The 384-well data were determined with a Labcyte Echo qualified 384-well acoustic grade polypropylene source plate (P-05525). The 1536-well data were determined with a Labcyte Echo qualified 1536-well, high-base, acoustic grade COC source plate (LP-03730).
Back to main list.What fluids can be transferred by the Echo liquid handlers?
ADE is a very effective way to transfer a wide variety of liquids from aqueous solutions to volatile solvents. Labcyte continues to develop algorithms to transfer a wide range of liquids. Please let us know if you have specific liquid that you want to use.
Back to main list.What type of plates can I use as destination plates?
The Echo Series 500 systems are compatible with a wide range of rigid, SBS-footprint destination plates. All destination plates must be at least 6.5 but no more than 14.4 mm in height and meet SBS specifications for low and medium flanges. One pharmaceutical company uses Echo technology to transfer 3456-well plates.
Many types of polystyrene plates have been used as destination plates. These plates are excellent for assays but are not recommended for long-term storage of undiluted DMSO due to incompatibility of DMSO and polystyrene.
Please contact Labcyte if you have any questions concerning your desired destination plate. We will check to see that flange heights, well geometries and other variables are consistent with current usage.
Back to main list.What types of compound storage plates are compatible with ADE?
All source plates in ADE must be of acoustic grade and Echo qualified. System precision and accuracy are guaranteed only with Echo qualified plates. These flat-bottomed plates are designed to provide optimal acoustic energy transfer for the best results. The table below shows the specifications for existing Echo qualified plates.
Unless otherwise specified, all Labcyte plates conform to the following SBS microplate standards.
ANSI/SBS 1-2004: Microplates - Footprint DimensionsANSI/SBS 2-2004: Microplates - Height DimensionsANSI/SBS 3-2004: Microplates - Bottom Outside Flange DimensionsANSI/SBS 4-2004: Microplates - Well PositionsSBS-5 Microplates - Side-Wall RigidityBack to main list.Can other source plates be added as a field upgrade?
Yes. As new source plates become available, you will be able to purchase the upgrades, as you need them. Your Labcyte representative will have the most information on current updates.
Back to main list.Who has purchased an Echo system?
Since the commercial release of the first member of the Echo Series 500 systems (the Echo 550 liquid handler) at least nine of the world’s top ten pharmaceutical companies have purchased systems. At least three universities have purchased systems and further systems have been bought by biotech and smaller pharmaceutical companies.
Back to main list.How much do Echo liquid handlers cost?
The systems come with a wide variety of options to meet the needs of our wide range of customers. Price also depends upon the geographical location of the user. Please contact your local Labcyte representative or click
here to contact someone who can explain the various options.
Back to main list.Can the Echo systems be put into an environmental control system?
The Echo 550 is compatible with many environmental conditions. It is designed to run optimally when the temperature ranges between 16-26ºC (61-83ºF) with relative humidity of <80%. Note that pure DMSO solidifies at 18ºC. For transfers, the DMSO must be in liquid form. DMSO:water, 95:5 (v:v) freezes at -10ºC.
Back to main list.Can the Echo systems detect crystals or precipitates in wells or tubes?
The Echo systems are not designed to detect crystals or precipitates. However, the systems will detect materials that affect the flow of acoustic energy in a well or tube. If crystals, bubbles, debris or precipitates occlude the acoustic beam, the system will detect and flag that there is an anomaly during the auditing process, and will not attempt to eject from that well. Any skipped wells are recorded in the transfer files and can be found by checking the server log file or opening the corresponding print.csv file. Note that if the crystals do not occlude the acoustic beam (and this is frequently the case), the system will eject droplets as though they were not present.
Back to main list.Why do the Echo liquid handlers require flat-bottomed plates?
To accurately and precisely eject a droplet from a well, the acoustic energy must be focused on the meniscus of the fluid. Containers that are not flat-bottomed act as lenses that change the acoustic focus. This dramatically decreases precision and accuracy and can lead to failed ejections.
Back to main list.Does the compound or analyte have an effect on transfer?
In general, there is little or no effect on transfer volumes when the concentration of the dissolved compound is between 4 and 8 mg/mL (or 10 mM for compounds with MW between 400 and 800). This poster "
Impact of Compound Concentration" shows that accuracy was affected by less than 2% and precision by less than 3% even when the solubility of the compound forced the material into a suspension. If precipitate collects at the bottom of the well and occludes the acoustic energy, the Echo system will skip the transfer from that particular well and record the anomaly.
Back to main list.The Echo systems transfer fluids into plates that are upside down! Won’t the fluids drain out?
For nanoliter droplets, surface tension is a much stronger force than gravity. In high-density plates the liquid is held tightly even when there are microliters of fluid in each well. Try the experiment yourself: put liquid into each well of a multi-well plate and turn it upside down. You will have to vigorously shake the plate or hit it on a table to knock the liquid out of the plate.
Back to main list.The Echo system movie shows a water column in the system. Doesn’t that increase DMSO hydration?
Sound waves travel more efficiently in water than in air. The water column couples the transducer to the sample. The short time for which a source plate is inside the instrument, the negative pressure on the water supply return line and the airflow patterns in the system all minimize hydration. The Echo 550 can easily monitor DMSO hydration over long periods, and we have determined that the hydration contribution from the water column is extremely low.
Back to main list.How do I know if an Echo Series 500 system has not ejected liquid from a particular well?
The GUI (graphical user interface) has an Advanced tab. By clicking on the appropriate box, the system will store data concerning skipped wells (due to underfill, overfill, or air bubbles in the well) so that the user can inspect the skipped wells. The system cannot distinguish between anomalies, leaving that determination to the user. This data can also be saved through the API when an Echo system is integrated into a robotics-based system. In this case, the user and the integrator must determine when and how they want to inform the user of missed wells.
Back to main list.What type of maintenance do the Echo systems require?
The Echo liquid handlers are capable of operating 24 hours per day, seven days per week (provided that the system is operated within the design constraints indicated in the standard Labcyte performance specifications.) Normal Customer and Preventative Maintenance Periods are as follows;
Customer Maintenance
Daily Maintenance .5 Hours
Weekly Maintenance .75 Hours
Biweekly Maintenance .75 Hours
Monthly Maintenance .75 Hours
Semi-Annual Maintenance 2.0 Hours
Labcyte Recommended Preventative Maintenance
Quarterly Maintenance 2.0-4.0 Hours
Semi-Annual Maintenance 6.0-8.0 Hours
Back to main list.Can Echo Series 500 systems be integrated into robotics-based systems? What integrators have you worked with?
Echo systems have been successfully integrated into robotics-based systems with internal integrators at pharmaceutical companies as well as with third-party integrators.
We have systems integrated via High Resolution Engineering (HRE), Process Analysis and Automation (PAA), RTS Life Sciences, Thermo CRS, and Velocity11.
Back to main list.The Portrait 630 Spotter
What fluids can be transferred with the Portrait reagent multi-spotters?
ADE is a very effective way to transfer a wide variety of fluids from aqueous solutions to volatile solvents. Labcyte continues to develop algorithms to transfer a wide range of liquids. Please let us know if you have a specific liquid that you want to use.
Back to main list.Deerac™ Liquid Handling Platforms
What are the dimensions of the capillary?
ID range: 0.0055” – 0.007” (0.14 – 0.18 mm); Nominal ID = 0.16 mm
OD range: 0.0120” – 0.0125” (0.30 – 0.32 mm); Nominal OD = 0.31 mm
Length: 1.125” +/- 0.01” (28.575 +/- 0.25 mm).
Back to main list.
What is the usable length of capillary? Can this be changed?
With spring-loaded alignment bar: 12 mm
Back to main list.What is the volume of the capillary?
575 nL
Back to main list.What is the volume of the entire tip?
Max internal tip volume = 350 µL
Back to main list.How much volume can be added to each channel using tubing?
0.75-1 mL per channel
Back to main list.How does aspirating into the tubing affect the dispense performance?
No difference in precision or accuracy.
Back to main list.Does dispensing cells/beads affect the tip performance; is there clogging?
The system has been validated for use with both yttrium oxide and polystyrene SPA beads and a variety of cells including bacterial cells and mammalian cell lines.
Back to main list.What is the expected lifetime of the tip?
The minimum expected lifetime is 250,000 dispense cycles. Typical lifetime is in the region of 1,000,000 cycles.
Back to main list.How can the firmware version be determined?
The DeeracTM setup utility will inform you in the “Setup Unit” function window.
Back to main list.How is the firmware updated in the field?
Platform control firmware: Requires replacement of the controller chip. Takes no more than 10 minutes.
Dispenser: The main firmware section can be re-programmed in the field through a serial cable with the electronics switched to update mode. Takes 20 minutes.
Back to main list.What are the standard liquid class settings for water? DMSO, etc?| Liquid | Pressure | Vacuum | Viscosity | Offset |
| Water | 1400 | 600 | 1000 | 0
|
| 20 % Glycerol | 1400 | 600 | 1760 | 0 |
| 30 % Glycerol | 2500 | 600 | 2000 | 0 |
| 40 % Glycerol | 3400 | 600 | 2900 | 0 |
| 50% Acetone | 1550 | 600 | 1000 | 0 |
| 90 % Glycerol | 1200 | 600 | 500 | 0 |
| 100% Acetone | 2500 | 600 | 1738 | 0 |
Back to main list.What effect does each liquid class parameter have on the dispense performance?
In summary:
- The pressure affects the dispenser precision (CV%). For a given liquid increasing the pressure will improve the CVs as the drops will form in a more regular manner. Note that beyond a certain point (liquid dependent) the CVs will begin to rise again.
- Altering the pressure should not affect the volume of liquid dispensed.
- The viscosity constant will affect the accuracy of the system for the liquid in question across the entire range by linearly scaling all volumes.
- For example, if for a given liquid with a set viscosity constant of, say 1000, requesting 500 nL gives 550 nL on average then altering the viscosity constant to 909 will scale the dispense characteristic so that requesting 500 will give 500.
- The vacuum level, which is applied during aspiration, should be left constant for each liquid.
- The offset is a volume which will be added to each volume request and may be used to shift the dispense characteristic up or down across the entire volume range.
- The difference in effect of the viscosity constant vs. the volume offset is shown below.
Back to main list.How often does the dispenser need to be recalibrated?
The dispenser does not have to be adjusted for precision. Degradation in CV normally indicates a worn-out tip.
It is recommended that the accuracy is tested every month as part of a user-defined performance-qualification.
Back to main list.What is the working volume?
The working volume is the volume of liquid aspirated into the tip along with the required dispense volume. It is the volume of liquid required to coat the magnet inside the tip and also provides liquid to compensate for the variation between aspirate and dispense accuracies. The working volumes is affected by aspirate volume, dispense volume, plate density (number of dispenses) and the viscosity of the liquid dispensed. As a result the working volume should be tuned for each assay.
Back to main list.What is the minimum working volume?
The minimum working volume is 3000 nL. This is the volume required to keep the magnet coated with liquid to provide a consistent dispense mechanism.
Back to main list.What are the external dimensions of the GX Series
Width = 388 mm
Depth = 670 mm
Height (including the head loom and guide wire) = 700 mm.
Back to main list.How often does the mechanics/robotics need to be re-calibrated?
The positional accuracy should be checked as part of the annual service procedure.
Back to main list.Can the pitch of the tips be changed for non-standard applications?
Custom development is possible but software and hardware modifications would be required incurring a development cost. At present 9mm is the only pitch available.
Back to main list.Is it possible to have a 2/4/12 tip system?
Channels are available in banks of 4 (4/8/12) but development of a non-8 channel system would be custom development for both hardware and software.
Back to main list.What is the resolution of the XY movement?
5 – 10 µm depending on speed
Back to main list.What is the resolution of the Z movement?
5 – 10 µm depending on speed
Back to main list.What is the travel range of the head?
The usable (i.e., head-addressable) range of motion on the DeeracTM liquid handlers is:
X: 204 mm
Y: 231 mm
Z: 50 mm
Back to main list.Can the reservoir/wash/plate positions be changed?
The standard Deerac GX Series bedplate is arranged to allow each tip on an 8-channel system to reach all wells of a 1536-well plate in both positions.
Positions of the bed elements may be altered for a custom requirement but all-tips-all-wells may not be possible under such a configuration.
What is the clearance of the tips over the bedplate?
Maximum distance between tips and bed plate = 50 mm
Back to main list.What is the volume of the reservoir?
Standard reservoir: 15 mL
Low dead volume reservoir: 13 mL
Back to main list.Can extra reservoirs be added?
The Deerac Control Software supports multiple reservoirs. Custom bedplate configurations are possible with multiple reservoirs; if required one wellplate may be replaced with extra reservoirs.
Back to main list.What operating systems are supported?
• Windows 2000
• Windows NT (see note below)
• Windows XP
Note that under NT a user must have administrator rights. A bug exists which prevents the spotting task being run correctly.
Back to main list.