Navigation Links
Precision Testing for the Sciclone Advanced Liquid Handler Z8 Option

Jeff Cunningham and Lynn Jordan
Caliper Life Sciences, Inc., Applications Laboratory, Hopkinton, Massachusetts, USA

Introduction

The Z8 liquid handling device is an optional accessory to the Sciclone Advanced Liquid Handler (ALH). This device is designed to permit individually and independently controlled access to single wells or columns of 96 and 384 well microplates for purposes of liquid transfer (tertiary dispensing). It can be configured to use fixed cannulas (1-50μL) or disposable tips (1-100μL) to facilitate standard hit-picking or re-formatting protocols. Here, we discuss the optimization of liquid transfer (1, 2, 5 and 10μL) techniques in terms of precision. Moreover, we extend the capabilities of the Z8 by performing continuous low volume (1-10μL) bulk dispensing across the columns of a 96 well plate (secondary dispensing) using fixed cannulas.

Materials and Methods

Instrumentation

All testing was performed using a Sciclone ALH with the Z8 option. Secondary dispensing is achieved using a Bulk Reagent Dispenser (Sciclone option) which facilitates the continuous delivery of water across the columns of a plate. Absorbance was measured at 450nm using a Wallac Victor2 Multilabel Counter (PerkinElmer).

Reagents

Tartrazine (SIGMA) solutions are prepared with deionized H2O and degassed with Helium at 0.7, 1.4, 5.0, and 6.0g/L) and dispensed at 10, 5, 2 and 1μL respectively.

Device Purging

The Z8 device is first purged to facilitate the elimination of compressible materials or gasses as follows:

  • Prime the lines of the Z8 until the system fluid reaches the lines just past the syringes.
  • Aspirate 100μL from source.
  • Manually remove trapped air in the syringe barrels by first loosening the plunger mounting screws then pushing up on each plunger 3-4 times quickly.
  • Re-prime the lines until liquid flows from the ends of the tip adapters or fixed cannulas.

As a result of the purging, the system fluid (in this case degassed and deionized water) forms a continuous path from its source, through pumps, lines and syringes to individual fixed cannulas or tip adapters.

Disposable Tip Precision Determination (1, 2, 5μL tertiary dispensing):

All testing was performed using the deck layout (Fig.1) and a protocol to execute the following:

  • Deliver 100μL of water into each well of a NUNC (Denmark) 96 well, clear bottom, untreated plate using the Bulk Dispenser.
  • Attach 100μL Automation Certified tips to the Z8 tip-adapters.
  • "Pre-wet" the tips by triturating 50μL of tartrazine solution.
  • Aspirate a 5μL air gap followed by 1, 2 or 5μL of tartrazine.
  • Dispense total content (tartrazine and air gap) into the first column of the 96 well plate and repeat the process for all columns.
  • Dispense 100μL of water into each well using the Bulk Dispenser.
  • Acquire and export absorbance data to ExcelTM for CV calculation.
  • Repeat for each of 5 plates at appropriate tartrazine concentrations.

Fixed Cannula Precision Determination (1, 2, 5 and 10μL tertiary dispensing):

All testing performed using the deck layout (Fig.2) and a protocol to execute the following:

  • Deliver 100μL of water into each well of a 96 well plate using the Bulk Dispenser.
  • Aspirate 10μL of air followed by tartrazine (1, 2, 5 or 10μL) and a final 5μL air gap.
  • Dispense total content (tartrazine and both air gaps) into first column of the 96 well plate.
  • "Quick wash" cannulas by priming 100μL into a waste reservoir twice then triturate 100μL of clean water three times.
  • Repeat previous four steps for all 12 columns.
  • Deliver 100μL of water into each well of a 96 well plate using the Bulk Dispenser.
  • "Thoroughly wash" cannulas by priming 100μL into a waste reservoir four times then triturate 100μL of clean water three times.
  • Acquire and export absorbance data to Excel for CV calculation.
  • Repeat for each of five plates at appropriate tartrazine concentrations

Fixed Cannula Precision Determination (1, 2 and 5μL secondary dispensing):
All testing performed using the deck layout (Fig.3) and a protocol to execute the following:

  • Deliver 100μL of water into each well of a 96 well plate using the Bulk Dispenser.
  • Aspirate air gap (10μL) followed by tartrazine solution (25μL) for a total of 35μL.
  • Dispense appropriate volume of tartrazine solution (1, 2 or 5μL) once into a waste reservoir then into each well of the test plate.
  • Deliver water (100μL) into each well with the Bulk Dispenser.
  • Acquire and export absorbance data to Excel for CV determination.< br>
  • Repeat for each of five plates at appropriate tartrazine concentrations.
  • Prime cannulas four times with water (100μL) into a waste reservoir.
  • Triturate cannulas for three cycles with water (100μL).

Results

Absorbance was measured and the data exported to ExcelTM for CV determination for five plates at each volume tested. The results of the dispense data for disposable tips (Table 1), fixed cannula secondary dispensing (Table 2) and fixed cannula tertiary dispense data (Table 3) are shown below. The overall CV's that comprise every well of a 96 well plate are shown for each volume tested. The mean %CV is caculated in each table.

Discussion

In this study, we explore the capabilities of the Sciclone ALH Z8 option using both disposable tips and fixed cannulas. We demonstrate its utility for tertiary dispensing (e.g., hit-picking), where small volumes of substance are transferred from single wells and dispensed into new destination wells. Moreover, we show that the functionality of the Z8 can be extended to perform low volume (1-10μL) secondary dispensing. This is noteworthy since the Bulk Dispenser option on the ALH is rated to dispense volumes of 10μL or greater. (see, Cunningham J and Jordan L "Sciclone ALH Bulk Dispense Precision Testing", Application Note-AP5, Zymark Corporation, 2001)

The precision results achieved in this study surpass the published factory specifications for disposable tips and fixed cannulas (CV <= 10% and 5%, respectively), at 1μL dispense volumes. While this is likely a result of good pipetting technique such as proper tip attachment and optimal aspirating and dispensing heights, we have found that purging the system of all air is equally important for proper Z8 functionality.


'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Sciclone ALH Bulk Dispense Precision Testing
2. 96 Well Pipetting Precision and Accuracy Using the RapidPlate 96/384 Workstation
3. 96 Well Pipetting Precision Testing Using the Sciclone ALH with a 384 Fixed-Cannula Array
4. Sciclone ALH 3000 Liquid Handler: Functional Characterization of the High Volume Head - Part 1: Plate-to-Plate Precision Testing
5. 384 Well Pipetting Precision and Accuracy Testing Using the Sciclone Advanced Liquid Handler
6. Reliable Assessment of Micronuclei for Genotoxicity Testing in Mice or Rats
7. Restriction Endonucleases Undergo Rigorous QC Testing to Ensure Optimal Performance
8. Bio-Plex Cytokine Immunoassays and ELISA: Comparison of Two Methodologies in Testing Samples From Asthmatic and Healthy Children, Rev A
9. Testing Compressed Air Lines for Microbiological Contamination
10. Testing Compressed Air Lines for Microbiological Contamination
11. Scalable Automation of the TempliPhi DNA Sequencing Template Amplification and DYEnamic ET Terminator Sequencing Kits on the Sciclone ALH 3000
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/23/2016)... ... 2016 , ... Mosio, a leader in clinical research patient ... and Retention Tips.” Partnering with experienced clinical research professionals, Mosio revisits the hurdle ... and strategies for clinical researchers. , “The landscape of how patients receive and ...
(Date:6/23/2016)... - FACIT has announced the creation of a ... Propellon Therapeutics Inc. ("Propellon" or "the Company"), to ... of first-in-class WDR5 inhibitors for the treatment of ... an exciting class of therapies, possessing the potential ... patients. Substantial advances have been achieved with the ...
(Date:6/23/2016)... ... June 23, 2016 , ... Charm Sciences, Inc. is ... has received AOAC Research Institute approval 061601. , “This is another AOAC-RI approval ... Bob Salter, Vice President of Regulatory and Industrial Affairs. “The Peel Plate methods ...
(Date:6/23/2016)... SAN FRANCISCO , June 23, 2016   ... it has secured $1 million in debt financing from ... to ramp up automation and to advance its drug ... for its new facility. "SVB has been ... goes beyond the services a traditional bank would provide," ...
Breaking Biology Technology:
(Date:6/16/2016)... 16, 2016 The global ... to reach USD 1.83 billion by 2024, according ... Inc. Technological proliferation and increasing demand in commercial ... to drive the market growth.      ... The development of advanced multimodal techniques for biometric ...
(Date:6/3/2016)... 3, 2016 Das ... Nepal hat ein 44 ... geprägter Kennzeichen, einschließlich Personalisierung, Registrierung und IT-Infrastruktur, ... Produktion und Implementierung von Identitätsmanagementlösungen. Zahlreiche renommierte ... Januar teilgenommen, aber Decatur wurde als konformste ...
(Date:5/24/2016)... patient care by providing unparalleled technology to leaders of the medical imaging industry.  As ... added to the range of products distributed by Ampronix. Photo - ... ... ... ...
Breaking Biology News(10 mins):