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Agilent Bravo Automated Liquid Handling Platform: 96-Channel 10 L Tip Accuracy Precision Manual

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1. the U S A April 2 2009 5990 3643EN Agg Agilent Technologies
2. Summary e Bravo 96ST head 10 uL tips e 0 3 ul 3 1 CV 1 5 accuracy e 0 5 uL 2 3 CV 0 8 accuracy e 2 0 uL 1 1 CV 0 8 accuracy e gt 2uL equal or better than 2 0 uL CV and accuracy Introduction Automated liquid handling devices are used in many processes performed in laboratories today Liquid handling automation increases sample processing and throughput and improves accuracy desired dispensed volume is equal to the actual dispensed volume and precision a narrow distribution of dispensed volume when compared to a person oper ating a handheld liquid pipettor Automated liquid handling devices can also significantly reduce the occurrence of errors in a process 2 To further the effort to automate routine laboratory processes Agilent Automation Solutions has developed the Bravo Automated Liquid Handling Platform Bravo Features The Bravo has nine deck positions which accommodate any 96 384 or 1536 well SBS standard microplates Deck positions can be configured with heating cooling and shaking stations as well as maintaining open locations for tip boxes sample microplates and reservoirs The Bravo can be used with 8 16 96 or 384 channel fixed tip or disposable tip heads Heads can be interchanged in a matter of seconds The Bravo is designed to be used either standalone or as part of a robotic laboratory automation system It is also designed to fit inside some models of laminar flow hoods
3. and retain laminar flow thereby opening up cell based plating and cellular assays to an automated liquid handling platform Agilent Bravo Automated Liquid Handling Platform 96 Channel 10 uL Tip Accuracy and Precision Technical Overview Accuracy and Precision Testing This technical overview describes a method to measure the accuracy and precision of the Bravo in conjunction with a 96 Channel ST Disposable Tip Head The 10 uL disposable tips were tested at the lower end of the practical volume range 0 3 2 uL Performance at volumes gt 2 uL meets or exceeds the 2 uL performance Measurements are determined by dispens ing a tartrazine DMSO solution into dry microplates filling the plates with water and measuring tartrazine absorbance The product s performance meets or exceeds CVs of 5 and accuracy of 10 of the desired volume across the practical volume range Materials e Bravo with a 96 Channel ST Disposable Tip Head e Agilent 384ST 10 uL tips product no 10734 202 e Agilent 96 well manual fill reservoir product no 08105 001 e 384 well polystyrene flat bottom plates Greiner 781101 e Tartrazine solution 0 25 w v dissolved in dimethylsulfoxide DMSO e UV Vis Spectrophotometer with a 405 nm filter Thermo Multiskan Ascent Method 60 mL of tartrazine solution is poured into a manual fill reservoir The reservoir is placed on position 2 of the Bravo A 10 uL tip box is placed on position 3 A 384 w
4. determine the precision of the dispense Coefficient of Variance CV calculations Accuracy 1 5 l l Outliers 5 7 Outliers 10 1 Results Volume 0 3 pL Precision CV 3 1 Transfer time 24 seconds plate were made by dividing the standard deviation by the mean The accuracy and precision results are typical based on averages of three successive dispenses Accuracy out liers are an average per plate Results may vary depending on individual experimental methods and liquid class optimization 0 5 pL 2 uL 2 3 1 1 0 8 0 8 3 0 1 0 24 27 Accuracy amp Precision of the 96ST Head with 10 uL Tips onthe Bravo Platform S L v o c amp a gt Accuracy m Precision Volume uL Plate Compatibility 96LT 250 uL Yes 384ST 10 uL Yes 384ST 30 uL Yes Tip Type 96 Well Microplates 96LT 200 uL Yes 384ST 70 uL Yes 384 Well 1536 Well Microplates Microplates Yes No Yes No Yes Yes Yes Yes Yes No eeso Conclusion In summary the liquid handling capabilities of the Agilent Bravo Automated Liquid Handling Platform in conjunction with a 96 Channel ST Disposable Tip Head provides precise accurate and consistent liquid transfers The 96 channel head combined with Agilent s 10 uL disposable tips can pipette across a low volume range 0 3 10 uL making it ideal for applications requiring precise dispensing at low volumes The highly configurable deck of the Bravo Automated Liq
5. ell polystyrene plate is placed on position 5 Agilent VWorks software liquid classes for 0 3 0 5 uL and The Agilent Bravo has nine deck positions and can be configured with interchangeable 8 16 96 or 384 fixed and disposable tip heads 1 5 uL dispenses are utilized A VWorks protocol is created and run in the following manner 1 Tips are pressed onto the head 2 0 3 0 5 or 2 0 uL tartrazine solution is aspirated from the manual fill reservoir Aspirate parameters are 6 mm from the bottom of the plate and preceded by a 0 4 uL pre aspirate volume 1 uL for 2 uL dispense 3 0 3 0 5 or 2 0 uL tartrazine solution is dispensed into one quadrant of a 384 well plate Dispense parameters are 0 mm from the bottom of the plate and fol lowed by dispensing a 0 4 uL blowout volume 1 uL for 2 uL dispense 4 The transfer is repeated for the other quadrants of the plate Dispensed volumes are diluted to 50 uL by the addition of water in this case with clean tips and a separate VWorks protocol Plates are centrifuged at 1800 rpm for 60 seconds to ensure full mixing and consistent well menisci Absorbance is read at 405 nm Eee Agilent Technologies Accuracy is calculated based on an equation derived from a tartrazine DMSO calibration Curve consisting of data points at 0 1 0 3 0 5 0 75 1 1 25 1 5 2 and 2 5 uL compared to the actual absorbance value in each well Absorbance values in each well are used to
6. uid Handling Platform along with its compact footprint makes it ideal for many applications in the laboratory including compound sample transfers genomic appli cations cell plating and cell based assays References 1 Sofia MJ Leveraging Compound Management Capabilities in Support of Drug Discovery From Sample Archive to Sample Distribution Driving Efficiency and Improving Productivity Laboratory Robotics Interest Group Meeting Oral Presentation Jan 2005 2 Holman JW Mifflin TE Felder RA Demers LM Evaluation of an automated preanalytical robotic work station at two academic health centers Clin Chem 2002 Mar 48 3 540 8 Please contact your sales representative or Agilent Applications Support if you have particular questions regarding your specific application Supplemental information pro tocol files and data analysis spreadsheets are also available upon request www agilent com lifesciences automation This item is intended for Research Use Only Not for use in diagnostic procedures Information descriptions and specifications in this publication are subject to change without notice Agilent Technologies shall not be liable for errors contained herein or for incidental or consequential damages in connection with the furnishing performance or use of this material Thermo and Multiskan Ascent are registered trademarks of Thermo Fisher Scientific Inc Agilent Technologies Inc 2009 Published in

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