Covaris R230 Manual

R230 Focused-ultrasonicator
Setup & Instrucon Manual
Instrument and Soware Developed for High-performance Sample Preparaon
using Adapve Focused Acouscs® (AFA®) Energy
For Research Only
Product Name: Revoluon | Model Numbers: R230 | Soware: SonoLab 10
R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020

2R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
UNIVERSAL PRECAUTIONS
Universal Precauons should be followed on all specimen samples, regardless of whether a sample is known to contain
an infecous agent. Laboratories handling specimen samples are advised to comply with applicable parts of the following
government and clinical standards, or their equivalent in the country of use:
• Centers for Disease Control (CDC), Universal Precauons for Prevenon of Transmission of HIV and Other Bloodborne
Infecons, published 1987, updated 1996
• Clinical and Laboratory Standards Instute (CLSI), GP17-A2 Clinical Laboratory Safety; Approved Guideline - Second
Edion, published 2004, ISBN 1-56238-530-5
• Clinical and Laboratory Standards Instute (CLSI), M29-A3 Protecon of Laboratory Workers from Occupaonally
Acquired Infecons; Approved Guideline, Third Edion, published 2005, ISBN 1-56238-5674
• Occupaonal Safety and Health Administraon (OSHA), 29 CFR 1910.1030 Bloodborne Pathogens
• Internaonal Standards Organizaon (ISO) 15190:2003, Medical Laboratories – Requirements for Safety
Trademarks
Patented or patent pending and registered or registraon-pending trademarks of Covaris are protected.
Registered names and trademarks used in this document, even when not specically marked as such, are not to be considered
unprotected by law.
WCS and AFA are either registered trademarks or trademarks of Covaris, Inc. in the United States and/or other countries.
Teon is a registered trademark of The DuPont Company.
Informaon subject to change without noce
For Research Use Only (RUO)
Not for use in Diagnosc Procedures
Publicaon P/N 010480
Product covered by Patent No. US6, 719,449, US6, 948,843, and other pending applicaons
Copyright © 2020 Covaris, all rights reserved.
Covaris, Inc.
14 Gill St, Unit H
Woburn, Massachuses
01801-1721 USA
Printed in U.S.A.

3R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
Warnings
For the Safety of Operang Personnel:
• Make sure that the equipment is properly grounded. DO NOT operate if it’s not properly grounded.
• The unit is equipped with a power plug appropriate for the desnaon country. DO NOT, under any circumstances, remove the
grounding prong from the power cord.
• DO NOT run an acousc treatment with the sample cover in the OPEN posion or without a water bath - the acousc system
will not work.
• If there is any indicaon that the Safety System is not funconing properly, DO NOT operate the equipment and contact Covaris
immediately.
• If the equipment is used in a manner not specied by the manufacturer, the protecon provided by the equipment may be
impaired.
To Prevent Damage to the Equipment:
• The instruments are designed to operate in ambient laboratory condions e.g., 19 °C to 25 °C (66 °F to 77 °C).
DO NOT operate the instrument in a cold room environment. The system is designed to operate with a water bath and re-
circulang heater/chiller apparatus to control sample temperature.
• NEVER run a method without water in the bath since this could damage the transducer. The instrument is equipped with a
water level sensor to protect the transducer and degassing pump. The system will not allow the degassing pump to operate or
the acousc treatment to start unless an adequate volume of water is detected.
• Dislled or deionized water should be used to ll the water bath.
• Unless the Water Condioning System 3 (WCS) is acvely employed, empty the water bath and wipe it dry EVERY DAY with a
lint-free cloth. DO NOT leave water in the tank for an extended me as there is no water ltraon or water cleaning system
within the apparatus (unless WCS is employed).
• DO NOT employ isopropyl alcohol, ammonia-based or abrasive cleaners in the system, as these will damage the acrylic surfaces.
• Establish a standard of operaon and test the equipment periodically, as described in the maintenance secon of this manual.
CAUTION: All WCS 3 components have pressure rangs above Covaris’ specied maximum operang pressure
which have been extensively tested. However, if plant DI water is connected to the WCS, a catastrophic failure
may result in substanal plant water spillage. It is the user’s responsibility to specify and implement appropriate
containment/alarm schema for their facility.

4R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
Warranty
When used in accordance with wrien instrucon and under normal operang condions, the Covaris instruments are guaranteed
to be free of defects in MATERIAL and WORKMANSHIP for one (1) year from the date of original delivery by an authorized
representave. Any component proven defecve during the stated period will be repaired free of charge or replaced at the sole
discreon of Covaris, F.O.B., Woburn, Massachuses, U.S.A. provided that the defecve component is returned properly packaged with
all transportaon charges prepaid. The customer is expected to perform basic diagnoscs and component replacement via telephone
support from Covaris personnel. If Covaris personnel are required to perform on-site repair, all travel related costs are paid by the
customer. A limited warranty as specied may apply to certain components of the equipment.
Warranty Excepons
This warranty is void if failure of the soware or hardware has resulted from accidents, abuse, improper maintenance or repair, or by
misapplicaon from the customer. It is also void if damage is caused by any unauthorized aachments or if modicaons are made to
the equipment. Removing or tampering with the Safety Enclosure will void the warranty, and the customer will assume all liabilies.
This warranty is limited to the original purchaser and is not transferable.
The soware will perform according to the accompanying wrien materials and the medium on which the soware is delivered is
free of defects in materials under normal use and service. The warranty is void if damage has resulted from third party soware not
intended for use with the system.
The high power focused transducer is designed to give maximal mechanical energy output in water. Permanent damage to the
transducer and electronic circuits could result if the transducer is operated without water. Operaon of the system without water in
the water bath voids the warranty.
CONTACT COVARIS, INC. SHOULD YOU HAVE QUESTIONS OR CONCERNS PERTAINING TO THE USE OF OUR EQUIPMENT.
Warranty Services
The purchased equipment is covered by a twelve (12) month warranty which includes all the necessary services and support necessary
so that the customer can successfully operate their equipment. Extended warranes are available at the end of the original 12 month
warranty period.
Services included with the original purchase of the system are:
Technical Support – Ongoing assistance with the operaon or applicaon of the equipment and/or troubleshoong is provided via:
• Telephone
- United States: Tel: [phone] during the hours of 9:00 a.m. to 5:00 p.m., Monday through Friday,
Eastern Standard Time (EST), Greenwich Mean Time (GMT-05:00)
- Europe: Tel: [phone], during the hours of 9:00 a.m. to 5:00 p.m., Monday through Friday, Greenwich Mean Time
• E-mail queries to [email protected] or applica[e-mail]
Parts Replacement – Replacement of parts (excluding consumables) from normal operaon of equipment are provided on a priority
basis. All labor and shipping charges are included. Failure due to accident, abuse, or improper operaon is not covered.
This manual includes maintenance guidelines and troubleshoong ps.

5R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
Table of Contents
1. Introducon ...................................................................................................................................................................................... 6
1.1. Overview of the User Manual ........................................................................................................................................... 6
1.2. The R230 Instrument and the Covaris Process .............................................................................................................. 6
2. Unpacking and Setup ...................................................................................................................................................................... 7
2.1. Unpack and Idenfy Components & Accessories. ......................................................................................................... 7
2.2. Install the On-Deck Module .............................................................................................................................................. 8
3. Geng Started ................................................................................................................................................................................. 17
3.1. Start the System ................................................................................................................................................................... 17
3.2. Start SonoLab ....................................................................................................................................................................... 17
3.3. System Shut-down .............................................................................................................................................................. 18
4. AFA Operaon .................................................................................................................................................................................. 19
5. SonoLab Soware ............................................................................................................................................................................ 20
5.1. The Method Screen ............................................................................................................................................................. 20
5.2. The Schedule Screen ........................................................................................................................................................... 25
5.3. The History Screen .............................................................................................................................................................. 28
5.4. The About Screen ................................................................................................................................................................ 30
5.5. The Admin Screen ................................................................................................................................................................ 30
6. Creang & Eding Treatment Methods ....................................................................................................................................... 36
6.1. Create a New Method ........................................................................................................................................................ 36
6.2. Using the Repeat Funcon ................................................................................................................................................. 42
6.3. Scanning Treatments ........................................................................................................................................................... 45
6.4. Using Drag and Drop with Treatment Steps ....................................................................................................................46
7. System Specicaons ...................................................................................................................................................................... 48
8. Maintenance ..................................................................................................................................................................................... 49
8.1. Cooling Air Intake ................................................................................................................................................................ 49
8.2. Water Bath ............................................................................................................................................................................ 49
8.3. Transducer ............................................................................................................................................................................. 50
8.4. WCS 3.................................................................................................................................................................................... 50
8.5. Safety System ....................................................................................................................................................................... 50
8.6. Degassing System ................................................................................................................................................................ 50
8.7. Cleaning the System ............................................................................................................................................................ 50
9. Troubleshoong ............................................................................................................................................................................... 51
Appendix A: Replace the WCS 3.0 Parculate Filter ..................................................................................................................... 52

6R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
1.1. Overview of the User Manual
This manual contains the operaon and maintenance instrucons for the Covaris R230 Focused-ultrasonicators. This document
contains informaon essenal to the proper use and care of the equipment.
Should any unforeseen problems occur with the normal operaon of the equipment, contact Covaris Technical Support immediately.
The following denions apply in this manual:
There is also a PDF version of this manual, typically available on the computer desktop. Open the manual by double clicking on its
icon.
1.2. The R230 Instrument and the Covaris Process
The R230 Focused-ultrasonicators are based on Covaris Adapve Focused Acouscs (AFA) technology. These instruments are
designed to provide an integrated tool to be used in a high-volume producon environment. This instrument is ideal for a variety of
applicaons including DNA shearing, mammalian cell lysis, low mass sample extracon from FFPE, ssue, whole blood, compound
screening, bead mixing, and pellet resuspension among other applicaons.
The system is comprised of the:
• Treatment System which delivers the acousc energy to the sample
• Safety System which protects users from inadvertently contacng the high intensity acousc energy
• Computer and Applicaon Soware which provides an interface to control the device via a standard user interface or API
• Water Management System, which consists of a cooler/heater and a UV treatment chamber with parculate lter to maintain
water quality in the instrument bath
The Covaris Process uses adapve focused acousc energy to precisely control cavitaon and acousc streaming within the sample
treatment vessel in a non-contact, isothermal fashion. Focused acousc energy is generated and delivered to individual samples. The
mechanical energy imparted on the sample results in a controlled series of compression and rarefacon events.
1. Introducon
NOTE: Inconvenience if disregarded.
CAUTION: Equipment damage may occur.
WARNING: Personal injury may occur.

7R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
2.1. Unpack and Idenfy Components & Accessories
The following items are included in the packaging found with the instrument:
• Electronics Box 1.0 (eBox)
• On-Deck R230 Focused-ultrasonicator 1.0 (for placement on the liquid handler deck)
• Water Condioning System 3.0 (WCS 3.0)
2. Unpacking and Setup

8R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
• Notebook Computer with AC Adapter. Most installaons will transfer Sonolab to the dedicated computer that is connected to
the Liquid Handler.
• Cables
- Computer AC adapter power cord
- Two instrument power cords
- Ethernet cable from eBox to Computer
- Communicaons and power cables between eBox, WCS 3.0, and On-Deck AFA
• Accessories
- The WCS Tubing Kit contains 3 hoses
- The Reservoir Kit contains 2 carboys and 2 hoses
- Mount adapter, sample tubes and racks (varies by applicaon)
- Setup kit
2.2. Install the On-Deck Module
2.2.1. Install the appropriate mounng/locang plate to the liquid handler by following the instrucons provided with the
mounng plate.
2.2.2. Remove the shipping locks from the On-Deck module.
There are 2 bolts that lock the On-Deck moon system in place for shipping. The bolts are accessed from the boom of the
base plate.
• Tilt the On-Deck module up to access the boom of the plate. In the image below, the module is resng on its back
edge with the 2 shipping bolts circled. Be careful to remove only these 2 bolts.
• Lower the On-Deck module so that it is again resng at on the bench.
NOTE: The notebook computer should be available for IQ/OQ, PM and Service. Please DO NOT use this computer for
other purposes.
NOTE: A mount plate must be used for all integraons with liquid handlers.
CAUTION: Shipping locks must be removed before powering up the instrument.

9R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
2.2.3. Store the shipping bolts on the On-Deck base plate.
• Remove the 5 at head cap screws, 2 from the right, and 3 from the le lower edge of the On-Deck cover. Do not
remove the screws in the inverted “U” shaped cut-outs.
• Li the cover o and set aside.
Remove these bolts only
Remove 5 screws

10R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
• Thread the two shipping bolts into the holes provided at the le rear of the module’s base plate, near the cable
connectors.
2.2.4. Level the On-Deck module in the liquid handler.
• Place the On-Deck module, without its cover, into the mounng plate installed previously in the liquid handler.
• Perform any nal adjustments required to the mounng plate.
• Level the On-Deck module by adjusng the 4 leveling thumbscrews, as necessary, while monitoring the 2 levels
provided on the base. There is a leveling screw at each corner of the module base. Three of the screws and the le-
to-right level are shown in the image of the module without the cover above. The front-to-back level is on the le side
of the base plate, between the cable and hose connector panels. See image below.
On-Deck Module with outer cover removed
Le-to-Right Level
3 of 4 Leveling Screws
Shipping Bolts

11R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
• Remove the On-Deck module from the liquid handler and reinstall the outer cover.
2.2.5. Complete installaon of the On-Deck module. Because of limited clearance in some liquid handlers, it is easier to connect
hoses and cables prior to placing the On-Deck module in the liquid handler.
• Connect the right-angle connectors of the “A”, “B”, and “DRAIN” hoses to the On-Deck module. The straight end of the
hoses can be connected aer the On-Deck module is in place on the liquid handler. Refer to secon 2.2.7 below.
• Connect the right-angle connector of the male 25-pin d-sub cable to the female 25-pin connector on the On-Deck module.
• Connect the right-angle connector of the male 44-pin d-sub cable to the female 44-pin connector on the On-Deck module.
• Connect the right-angle connector of the coax cable to the BNC connector on the On-Deck module.
Front-to-Back Level

12R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
• Place the On-Deck module with cables and hoses aached into the mounng plate on the liquid handler.
2.2.6. Arrange the eBox and WCS 3.0 as shown. These may be stacked on the oor or an adjacent table. The eBox may be
mounted on top of the WCS 3.0 as shown below. Use the two M4 x 25 buon head cap screws that come in the
accessories box to secure the eBox to the WCS 3.0.
Cable connecons
to the On-Deck
Module
CAUTION: When posioning the system, please keep the fan and rear panel vents free of obstrucons that may block
the ow of air.

13R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
2.2.7. Complete the water connecons.
There are two hose kits included with the R230. The WCS Tubing Kit is used to connect the On-Deck to the WCS and to
the Waste reservoir. The Reservoir Kit includes the Waste and Supply carboys and the two hoses which connect the WCS
to the carboys. From the WCS Tubing Kit:
• Connect the straight end of the “A” hose to the WCS “A” port. The “A” hose has a yellow tag at each end.
• Connect the straight end of the “B” hose to the WCS “B” port. The “B” hose has smaller ngs than the “A” hose.
• Connect the straight end of the remaining, clear “SPILL WAY DRAIN” hose to the port on the side of the waste
reservoir cap. See next three images.
Straight ngs at this
end connect to the WCS.
Angle ngs at
the end connect
to the On-Deck.

14R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
• Connect the right angle end of the clear hose to the “SPILL WAY DRAIN” port of the On-Deck.
Angle ng connects
to the On-Deck
“SPILL WAY PORT”
Straight ng connects
to the port on the side of
“Waste” carboy cap
To “SPILL WAY DRAIN”
of On-Deck
To “Waste” port of WCS
Waste Carboy with Hose Connecons

15R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
From the Reservoir Kit:
• Connect one hose from the WCS “Waste” port to the top port of the waste carboy’s cap.
• Connect the remaining hose from WCS “Supply” port to the supply reservoir cap, or another source of dislled or
deionized laboratory water.
The WCS contains a UV sterilizer and parculate lter to retard biological growth.
The middle connecon is used only during monthly maintenance procedures (see secon 8.2.2 for details on monthly
maintenance). No hoses should be connected to the maintenance connector while running the R230.
2.2.8. Complete the electrical connecons from the On-Deck Module:
• Connect the eBox female 25-pin connector, labeled “OD25”, to the On-Deck female 25-pin connector.
• Connect the eBox female 44-pin connector, labeled “OD44”, to the On-Deck female 44-pin connector.
• Connect the eBox BNC connector to On-Deck BNC connector.
Hose Connecons at rear of On-Deck
To “B” Port to WCS
To “Waste” Carboy
To “A” Port to WCS

16R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
2.2.9. Complete the electrical connecons from the WCS 3.0.
• Connect the eBox male 25-pin connector, labeled “WCS25”, to the WCS male 25-pin connector.
• Connect the eBox male 15-pin connector, labeled “WCS15”, to the WCS male 15-pin connector.
• Plug the WCS 3.0 into a power source using one of the line cords. The WCS 3.0 is a “universal input” device and
automacally employs any worldwide line voltage or frequency (100 to 240 volts AC, 50 or 60 Hz). Leave power
switched OFF at this me.
2.2.10. Complete electrical connecons from the eBox and laptop computer.
• Connect either eBox ethernet port to the ethernet port on the laptop computer. The R230 uses 4 xed addresses (5,
7, 10, and 22) on the 192.168.0 subnet.
• Connect the computer to a power source. Connect the mouse or other accessories to the computer.
• Plug the eBox into a power source using one of the line cords. The eBox is a “universal input” device and automacally
employs any worldwide line voltage or frequency (100 to 240 volts AC, 50 or 60 Hz). Leave the instrument power and
notebook power switched OFF at this me.

17R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
3.1. Start the System
• Power on the eBox using the power switch located on the rear, right side of the eBox.
- This provides power to the On-Deck module.
• Power on the WCS 3
- SonoLab soware will perform all temperature control.
3.2. Start SonoLab
Wait 30 seconds aer powering on the WCS 3, to allow the WCS 3 to inialize. Locate the Sonolab 10 icon on the laptop’s Desktop.
Double click on the icon. SonoLab will load and the user is prompted to home the R230 moon system.
Click “Ye s”, then “OK” when prompted to empty the sample holder and the On-Deck module will go through a moon homing process.
When homing is complete, SonoLab will display the Method screen. The Method screen displays the last successful treatment method
run and provides access to status informaon and basic instrument control funcons. See image below.
3. Geng Started

18R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
SonoLab Method Screen
The rst me SonoLab is started, remove the desiccant bag from the On-Deck module and ll the On-Deck bath with deionized water:
• From the Method screen, click on the Moon Control buon at the right side of the screen.
The Moon Control panel is displayed.
• Click the Open Cover buon. The On-Deck safety covers will open.
• Remove or cut the plasc wrap securing the desiccant bag to the On-Deck sample tray and remove the desiccant bag.
• With the desiccant bag removed, manually pour approximately 1 liter, or a bit more, of deionized water into the water bath to
prevent ow errors when the WCS 3 is powered on for the rst me. It is only necessary to do this the rst me the R230 is
started, when the water system is completely empty.
• Click the Close Cover buon in the Moon Control panel.
The R230 is now ready to use. The automaon computer can connect and assume control of the R230, or treatment methods may be
run directly from the Method screen.
See Secon 5 below for a more detailed descripon of SonoLab.
3.3. System Shutdown
If the instrument will be idle for an extended period of me, refer to the system shutdown procedure in the User Maintenance secon
of this manual.

19R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
AFA uses four parameters to ne tune specic acousc treatments. These parameters are programmed into Methods, provided by
Covaris. Methods can be executed manually or by the automaon soware, including specifying the treatment to be performed and
whether a full or only part of a plate is to be treated. A brief explanaon of the treatment parameters is provided below.
Peak Incident Power (PIP)
This parameter measures the peak electrical power used to drive the ultrasound transducer. In the R230, the range of Peak Incident
Power is 1.5 to 450 Was.
Duty Factor
This parameter denes the rao between transducer on-me and total operaon me. Duty Factor is expressed as a percentage. For
a given Peak Incident Power, a larger Duty Factor results in more acousc energy generated from the transducer. Users can dene the
Duty Factor from 2 to 50% in the R230.
The product of the Peak Power and Duty Factor gives the approximate Average Incident Power (AIP) generated from the transducer.
For example, if Peak Incident Power is set to 10 Was and Duty Factor is set to 15%, the Average Incident Power will be approximately
1.5 Was. However, the real average incident power from the transducer may vary due to factors such as sample tube type, water
level, and water temperature.
Cycles per Burst
This parameter indicates the number of electrical signals used in each acousc “burst” of the transducer. Cycles per Burst (cycles/
burst) range from 50 to 1000 in the R230 for dierent applicaons.
Duraon
This parameter denes the amount of me for which the sample is under treatment while in indexed operaon, i.e. treang one column
at a me. The measurement of Duraon is in seconds. For scanning treatments, the sample tray is passed connuously over the
transducer. In this mode, treatment me is a funcon of scanning speed. Scan speed ranges from 0.1 to 40 millimeters/second in the
R230.
Treatment Records
SonoLab History contains a record of parameter informaon for each treatment method run. An RFID scan is performed on each
treated sample tray to ensure that the correct consumables are being used for the treatment and to provide informaon for LIMS.
4. AFA Operaon

20R230 Setup & Instrucon Manual
Covaris
Part Number: 010480 Rev C | August 2020
5.1. The Method Screen
When SonoLab is launched, the Method screen is displayed with the last successfully run treatment method.
• Method displays the current selected treatment method and its status.
- Plate: the name of the sample plate specied by the current selected method.
- Temperature: the actual temperature of the water in the On-Deck bath.
- Setpoint: the On-Deck bath temperature specied by the current selected method.
- State: possible states, depending on what the R230 system is doing, includes:
○ At inial startup, Idle.
○ During a treatment method, the possible states include:
> RFID Scanning: the system is scanning and validang the RFID tag on the current treated sample plate.
> Waing for Water Level: the system waits for the water to reach the level specied by the selected method.
> Waing for Water Temperature: the system waits for the water temperature to be within the range specied by
the selected method.
> Waing for Water Degas: the system waits for sucient me for eecve degassing to occur.
> Running: the system is currently performing the selected treatment method steps.
> Paused: the method is paused by the user or the API.
5. SonoLab Soware