Application Note

Enabling Automated Microplate
Replication Using the Agilent Bravo
BenchCel WorkstationAuthors
Manuel A. Gomez Jr. andChad M. Whitman
Agilent Technologies, Inc.
Santa Clara, CA, USA

Introduction
Microplate replication is one of the most common workflows used in compound
management research facilities. Because of the large quantity of microplates thatmust be processed daily, it is a labor-intensive task ideally suited for automation.
In addition to increasing throughput, automation itself provides added consistency
and reliability.
The Agilent Bravo Automated Liquid Handling Platform is equipped to handle a wide
range of automated workflows, including miroplate replication. It has nine standard
deck positions that can hold any ANSI-standard microplate, and be customized witha variety of accessories to enable functionalities such as heating, cooling, shaking,barcode reading, and tip washing. The Agilent Bravo Pipetting Head can enable
volume transfers by column or row, from 300 nL to 200 µL. The integration of theAgilent BenchCel Microplate Handler with the Bravo provides an efficient wayof storing and dispensing microplates and pipette tip boxes, permitting users toprocess multiple batches of microplates with more walk-away time. This Application
Note describes how these two integrated instruments, combined as an Agilent Bravo
BenchCel Workstation, can provide a complete workflow solution for automating
microplate replication. Instrumentation is driven by the Agilent VWorks Automation
Control Software using a simplified interface that is easy to use and fully enabled forthis application.

Experimental
Equipment
The microplate replication workflow
was carried out using an Agilent
Bravo BenchCel Workstation System
consisting of:•Agilent BenchCel 4R Microplate
Handler with 660 mm Front
Loading Racks•

Agilent Bravo Automation Liquid
Handling Platform with Gripper•

Agilent 96LT Disposable PipetteTip Head•

Agilent 96LT 250 μL Disposable
Pipette Tips•

96 well polystyrene, clear flat bottom
microplates (Greiner 655101)•Nunc 96-deep wellpolypropylene, U bottom plates(ThermoFisher 260251)

Figure 1. Agilent Bravo BenchCel Workstation System for AutomatedMicroplate Replication.

Agilent VWorks Automation
Control Software
To simplify startup for new users,
preloaded form-based protocols forautomated microplate replication areprovided with each Bravo BenchCel
Workstation. Protocol variations for each
BenchCel Configuration are included.

Microplate Replication Protocol
Instrument setup
1.

Open VWorks Automation Control
Software and load the Plate
Replication System Form (Figure 2).

2.

Load Source microplates into
BenchCel Stacker 3.

3.

Stack Destination plates onto Bravo
deck position 4.

4.

Load pipette tip boxes into BenchCel
Stacker 1.

2

Figure 2. Agilent VWorks Protocol Form for Automated Plate Replication Systems.

5.

In the Pipette Settings
submenu, enter the desiredpipetting parameters.

6.

In the Run Setup submenu, enter
the desired number of Source
microplates under Plates to Process
and Destination microplates
under Plate Replicates. Select thelabware type used for Source andDestination microplates.

7.

Select Start Run to begin
automated workflow.

Instrument workflow
1.

Transfer pipette boxes from
BenchCel Stacker 1 to Bravo deckposition 2.

2.

Transfer source microplates from
BenchCel Stacker 3 to Bravo deck
position 6.

3.

Transfer destination microplates
from Bravo deck position 4 to Bravo
deck position 5.

4.

Transfer solutions from source
microplates to destination
microplates using set dispensing
and replicating parameters. Pipettetips are replaced for each source
microplate used in the protocol.

5.

Upon completion, source microplateis transferred back to BenchCel
Stacker 4.

6.

Destination microplates are stacked
vertically on Bravo deck position 8.

7.

Used pipette tips are re-racked into
their tip boxes and transferred backto BenchCel Stacker 2.

Results and Discussion
Microplate replication is astraightforward process in which definedquantities of a solution are transferredfrom a single source microplate intomultiple destination microplates(Figure 3). These destination plates
are then used for multiple downstream
applications such as small molecule
screening in research or toxicitiy studies.
Having an automated workflow for thisroutine task enables greater consistency,
while increasing walk-away time.

While a stand-alone Bravo Liquid
Handling Platform is capable ofperforming microplate replication, there
are limitations for high-throughput
worflows. Integration with the BenchCel
Microplate Hanlder significantly
increases the microplate capacity ofthis protocol by simultaneous andindependent transfer of microplates
and pipette tip boxes at Bravo deckposition 3.

SourceFive replicates

Figure 3. Microplate Replication
Workflow Example.

To demonstrate this workflow, anautomation microplate replication
workstation was assembled using theBravo Liquid Handling Platform with
the 96LT Channel Pipette Head andthe BenchCel 4R Microplate Handler.
The Bravo Liquid Handler included agripper attachment to move microplates
between various deck positions. The
96-channel pipetteing tip head was used
to handle Agilent 250-µL disposable
pipette tips. Transfer and storage ofmicroplates or pipette tip boxes wasperformed by the BenchCel 4R.

The Bravo BenchCel Workstation
control was handled by Agilent VWorks
Automation Control software using
a form-based interface designed forautomating microplate replication.
Source microplates can be replicated ina variety of ratios into new Destination
plates using the adjustable fields foundin the Pipette Settings and Run Setup
submenus. To ensure reproducibility andminimize contamination, pipette tips aredesigned to be changed after each set ofdestination microplates is produced.

3Conclusion
The Agilent Bravo BenchCel Workstation
provides a complete liquid handlingand microplate management
solution for enabling automatedmicroplate replication workflows. The
Agilent BenchCel Microplate Handler
facilitates simultaneous deliveryof Source microplates, Destination
microplates, and pipette tips to theAgilent Bravo Liquid Handling Platform.
Inclusion of a protocol form in the
Agilent VWorks Automation Control
Software enables quick start-up time
and the ability make simple protocol
adjustments to achieve customizablesource-to-destination microplate ratios.
Depending on the number of plates toprocess, and the pipetting parameters
used, it takes approximately 7 minutes
generate five replicate plates from one
Source plate and one pipette tip box
on a Bravo BenchCel Workstation.
Additionally, these forms can be used
as a template to build more customized
workflows, when the workstation is
integrated with additional devices onto
the BenchCel Microplate Handler. Overall,the Bravo BenchCel Workstation provides
a simplified method for automating
microplate replication to meet theneeds of high-throughput screening
research facilities.

www.agilent.com/chem
For Research Use Only. Not for use in diagnostic procedures.
DE.5911458333
This information is subject to change without notice.
© Agilent Technologies, Inc. 2017, 2020
Printed in the USA, October 29, 2020
5991-8092EN