Covaris LE220 Manual

Covaris
LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
Part Number: 010398 Rev H | November 2019
LE220 (-plus, R-plus, and Rsc)
Setup & Instruction Manual
Instrument and Software Developed for High-performance Sample
Preparation using Adaptive Focused Acoustic
®
(AFA
®
) Energy
For Research Only
Model Numbers: LE220-plus, LE220R-plus, and LE220Rsc | Software: SonoLab 8

Covaris
LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
Trademarks
Patented or patent pending and registered or registration-pending trademarks of Covaris are protected.
Registered names and trademarks used in this document, even when not specifically 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. Teflon is a
registered trademark of The DuPont Company.
Information subject to change without notice
For Research Use Only (RUO)
Not for use in Diagnostic Procedures Publication
PN 010398
Revision H, November 2019
Product covered by Patent No. US6, 719,449, US6, 948,843, and other pending applications 2019©
Covaris Inc., All Rights Reserved
Covaris, Inc.
14 Gill St, Unit H Woburn,
Massachusetts 01801-
1721 USA
UNIVERSAL PRECAUTIONS
Universal Precautions should be followed on all specimen samples, regardless of whether a sample is known to
contain an infectious 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 Precautions for Prevention of Transmission of HIV and Other Bloodborne
Infections, published 1987, updated 1996
•Clinical and Laboratory Standards Institute (CLSI), GP17-A2 Clinical Laboratory Safety; Approved Guideline - Second
Edition, published 2004, ISBN 1-56238-530-5
•Clinical and Laboratory Standards Institute (CLSI), M29-A3 Protection of Laboratory Workers from Occupationally
Acquired Infections; Approved Guideline, Third Edition, published 2005, ISBN 1-56238-5674
•Occupational Safety and Health Administration (OSHA), 29 CFR 1910.1030 Bloodborne Pathogens
•International Standards Organization (ISO) 15190:2003 Medical Laboratories – Requirements for Safety

Covaris
LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
Warnings
For the Safety of Operating 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 destination country. DO NOT, under any circumstances, remove the grounding
prong from the power cord.
•DO NOT run an acoustic treatment with the sample cover in the OPEN position or without a water bath - the acoustic system will not
work.
•If there is any indication that the Safety System is not functioning properly, DO NOT operate the equipment and contact Covaris
immediately.
•If the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired.
To Prevent Damage to the Equipment:
•The instruments are designed to operate in ambient laboratory conditions e.g., 19 to 25 °C (66 to 77 °F).
DO NOT operate the instrument in a cold room environment. The system is designed to operate with a water bath and re- circulating
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 acoustic
treatment to start unless an adequate volume of water is detected.
•Distilled or deionized water should be used to fill the water bath.
•Unless a Water Conditioning System 3.0 (WCS) is actively 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 time as there is no water filtration 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 operation and test the equipment periodically, as described in the maintenance section of this manual.
CAUTION: All WCS 3 components have pressure ratings above Covaris’ specified maximum operating pressure
which have been extensively tested. However, if plant DI water is connected to the WCS, a catastrophic failure
may result in substantial plant water spillage. It is the user’s responsibility to specify and implement appropriate
containment/alarm schema for their facility.

Covaris
LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
Warranty
When used in accordance with written instruction and under normal operating conditions, 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 representative. Any
component proven defective during the stated period will be repaired free of charge or replaced at the sole
discretion of Covaris, F.O.B., Woburn, Massachusetts, U.S.A. provided that the defective component is returned properly packaged with all
transportation charges prepaid. The customer is expected to perform basic diagnostics 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 specified may apply to certain components of the equipment.
Warranty Exceptions
This warranty is void if failure of the software or hardware has resulted from accidents, abuse, improper maintenance or repair, or by
misapplication from the customer. It is also void if damage is caused by any unauthorized attachments or if modifications are made to the
equipment. Removing or tampering with the Safety Enclosure will void the warranty, and the customer will assume all liabilities.
This warranty is limited to the original purchaser and is not transferable.
The software will perform according to the accompanying written materials and the medium on which the software is delivered is free of
defects in materials under normal use and service. The warranty is void if damage has resulted from third party software 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. Operation 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 warranties 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 operation or application of the equipment and/or troubleshooting 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 techsuppo[email protected] or [e-mail]
Parts Replacement – Replacement of parts (excluding consumables) from normal operation of equipment are provided on a priority basis.
All labor and shipping charges are included. Failure due to accident, abuse, or improper operation is not covered.
This manual includes maintenance guidelines and troubleshooting tips.

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LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
Table of Contents
1. Introduction .................................................................................................................................................................................... 6
1.1 Overview of the Instruction Manual ....................................................................................................................................6
1.2 The LE220-plus Instrument................................................................................................................................................... 6
1.3 The LE220R-plus Instrument ................................................................................................................................................6
1.4 The LE220Rsc Instrument .....................................................................................................................................................6
2. Installation and Setup ..................................................................................................................................................................... 7
2.1 Assembly............................................................................................................................................................................... 7
3. Getting Started................................................................................................................................................................................ 11
3.1 Fill the Water Bath................................................................................................................................................................ 11
3.2 Start the System ................................................................................................................................................................... 11
3.3 Preparing your System for Use ............................................................................................................................................. 13
3.4 Running a Method................................................................................................................................................................ 22
3.5 Daily System Shutdown ........................................................................................................................................................ 23
4. Create and Edit Methods ................................................................................................................................................................ 24
4.1 Create New Method ............................................................................................................................................................. 24
4.2 Creating Methods using Repeat Function ............................................................................................................................ 30
4.3 Scanning Treatments (LE220Rsc Only) ................................................................................................................................. 33
4.4 Modifying Existing Methods................................................................................................................................................. 34
5. SonoLab™ Application Software ..................................................................................................................................................... 35
5.1 Home Screen ........................................................................................................................................................................ 35
5.2 History Tab ................................................................................................................................................................................................ 35
5.3 About Tab .................................................................................................................................................................................................. 37
5.4 Admin Tab ................................................................................................................................................................................................. 38
6. System Specifications...................................................................................................................................................................... 42
7. Maintenance ................................................................................................................................................................................... 43
7.1 Cooling Air Intake ................................................................................................................................................................. 43
7.2 Water Bath ........................................................................................................................................................................... 43
7.3 Transducer............................................................................................................................................................................ 44
7.4 Safety System ....................................................................................................................................................................... 44
7.5 Degassing System ................................................................................................................................................................. 44
7.6 Cleaning the System ............................................................................................................................................................. 44
7.7 System Shutdown................................................................................................................................................................. 44
8. Troubleshooting .............................................................................................................................................................................. 46
9. Appendix ......................................................................................................................................................................................... 48
Appendix A: LE220R-plus (LE220-plus) Series Site or Transport Preparation ............................................................................. 48
Appendix B: Installing SonoLab 8 and Required Support Software ............................................................................................ 49
Appendix C: Humidity and Water Management for the Covaris LE220-plus .............................................................................. 54
Appendix D: Connecting AutoFill to Plant Water ........................................................................................................................ 56
Addendum for LE220Rsc...................................................................................................................................................................... 57

Covaris
LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
1. Introduction
1.1 Overview of the User Manual
This manual contains operation and maintenance instructions for the Covaris LE220-plus, LE220R-plus, and LE220Rsc Focused- ultrasonicators. This
document contains information essential to the proper use and care ,of your equipment. Should any unforeseen problems occur with the normal
operation of the equipment, contact Covaris Technical Support immediately.
The following definitions apply in this manual:
NOTE: Inconvenience if disregarded. WARNING: Personal injury may occur. CAUTION: Equipment damage may occur.
There is also a PDF version of this manual, typically available on the computer desktop. Open the manual by double clicking the user manual
icon.
1.2 The LE220-plus Instrument
The LE220-plus Focused-ultrasonicators are based on Covaris Adaptive Focused Acoustics (AFA) technology. These instruments are designed to
provide researchers an integrated stand-alone tool to be used in a laboratory environment. The instrument is ideal for those who work with
tissue homogenization, cell lysis, DNA or chromatin shearing, and other sample preparation steps.
The system is comprised of the:
•Treatment System which delivers the acoustic energy to the sample
•Safety System which protects users from inadvertently contacting the high intensity acoustic energy
•Computer and Application Software which provides an interface to control the device via the API
•Water Management System; which consists of a cooler/heater and a UV treatment chamber with particulate filter to maintain water
quality in the instrument bath
1.3 The LE220R-plus Instrument
In addition to the features of the LE220-plus described above, the LE220R-plus is designed to be integrated with a sample handling robot in an
automated work cell. The front door of the safety enclosure contains a cutout providing the robot arm access to the inside of the LE220R-plus
cabinet. The LE220R-plus presents the sample tray at the opening, and the robot arm loads or unloads the sample through the opening. The
LE220R-plus is controlled by custom software built using the Covaris Integration Toolkit.
A light curtain has been added as an additional safety interlock. The light curtain is wired into a safety rated relay controller the output of
which is connected to the Covaris controller board. If an operator reaches through the robot access opening while the system is running, motor
power and acoustics are immediately disabled.
The light curtain consists of invisible, infrared light and an infrared detector. Beams of light on 14 mm spacing are transmitted between the
source and detector. Blocking one or more beams of light trigger the safety interlock system. Acoustics and motion will remain paused until
the beams are unobstructed.
1.4 The LE220Rsc Instrument
The LE220Rsc is an enhanced version of the LE220R-plus. In addition to the features of the LE220-plus and LE220R-plus, the LE220Rsc
contains an extended travel X axis motion stage. This longer travel stage allows treatment of 96 and 384 position well plates using the
Covaris scanning treatment method.

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LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
2. Installation and Setup
2.1 Assembly
The following items should be found in the crate containing the instrument:
•Main unit
•Notebook computer with AC adapter
•Cables
-USB cable
-Instrument power cord
-Computer AC adapter power cord
•Accessories
-AFA ultraCUBE, consisting of:
○Water Conditioning System 2.0 (WCS)
○CH05 Chiller
-Chiller hose kit
-Sample tubes & racks (varies by application)
-Water tank (installed in main unit)
-AutoFill accessory kit
LE220-plus Focused-ultrasonicator LE220R-plus Focused-ultrasonicator
2.2.1 Unpack the instrument, the Water Conditioning System 2.0 (WCS), and the CH05 chiller. Refer to the WCS User Manual to install
the quartz tube and UV lamp into the WCS. The quartz tube and lamp are shipped separately to prevent breakage during shipment.
Arrange the instrument, WCS, and chiller on a bench as shown below. The chiller and WCS may be installed on either side of the
system. The CH05 and WCS may also be placed on the floor, if they can be located such that the hoses reach the front of the water
bath.
CAUTION: When positioning the system, please keep the fan and rear panel vents free of obstructions that may block
the flow of air.

Covaris
LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
Connect the hoses between the WCS and the CH05 as shown above.
Connect the remaining two hoses between the two ports on your LE220R-plus or LE220-plus water bath and the remaining two ports on the
opposite side of the WCS. It doesn’t matter which bath port connects to which WCS port for these two hoses.
The WCS contains a particulate filter and UV lamp to prevent biological growth. This allows the water bath to be changed just once per
month. Refer to the User Maintenance section of this manual for details.
NOTE: See the addendum at the end of the manual for connecting the hoses to the LE220Rsc water bath. The
connections are not interchangeable on the LE220Rsc.

Covaris
LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
2.2.2 The AutoFill option allows plant deionized water to be plumbed directly to the LE220R-plus and LE220-plus to allow automatic
filling of the water bath without any operator intervention. If the AutoFill option is going to be used, refer to the instructions
included with the AutoFill accessory kit or Appendix D of this manual to assemble the components of the AutoFill Accessory Kit, and
connect plant water to the system.
2.2.3 Plug the instrument into a power source using the enclosed line cord. The instrument is a “universal input” device and automatically
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 time.
2.2.4 Plug one end of the USB cable into the instrument USB Port and the other end into any USB port on the notebook computer.
2.2.5 Connect the temperature control cable between the D-connector at the rear of the LE220-plus or LE220R-plus and the D-
connector on the CH05 chiller.
2.2.6 Connect the CH05 to a power source. The CH05 is a universal input device. Do not turn on the CH05 at this time.
2.2.7 Locate the WCS power supply and line cord. Connect the WCS power supply to the port provided on the WCS and connect the line
cord to a power source. The power supply is a universal input device. Leave the WCS turned off at this time.
2.2.8 Connect the computer to a power source. Connect the mouse or other accessories to the computer.
2.2.9 Refer to the rear of the instrument’s right-side panel and identify the following features as necessary:
The Rear Right Panel of the System
AC Line (Mains) Input
and Power Switch
USB Port
Serial Tag,
Regulatory, and
Line Power Label
Product Safety Label

Covaris
LE220 (-plus, R-plus, and Rsc) Setup & Instruction Manual
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Part Number: 010398 Rev H | November 2019
2.2.10 Refer to the figure below and identify the following items:
LE220R-plus LE220-plus
2.2.11 Power on the instrument using the power switch at the right rear, at the power entry module.
2.2.12 Press and hold the green ‘Door’ button while pulling the door handle to open the safety enclosure front door. Slide out the water tank
and remove it from its plastic shipping bag. Remove and discard the desiccant pack.
The instrument is now ready to be started.
Enclosure Door
Safety Lock
Enclosure
Door Handle
Door Lock
Release Button
Stop Button
Safety Enclosure
Water Tank