Navigation Links
96 Well Pipetting Precision Testing Using the Sciclone ALH with a 384 Fixed-Cannula Array

Rudy Willebrords1 and Bart Moyson2
1Johnson & Johnson Pharmaceutical Research & Development, a division of Janssen Pharmaceutica N.V., Laboratory of Virology, Beerse, Belgium
2Caliper Life Sciences Benelux NV, Teralfene, Belgium

Introduction

The purpose of this paper is to report the performance obtained by dispensing 1μL organic sample (DMSO) containing tartrazine into 384 well plates containing water. Precision and accuracy results are shown for this work.

Instrumentation
All testing was performed using a Sciclone Advanced Liquid Handler (ALH) with a 384 Fixed-Cannula Array (Caliper Life Sciences). A Bulk Reagent Dispenser (Caliper Life Sciences) facilitated the continuous delivery of water across the columns of a 384 Well, Flat Bottom, Untreated, Polystyrene Assay plates (COSTAR Catalogue # 9837). Absorbance was measured at two wavelengths using a SPECTRAmax (Molecular Devices Corporation) 2000/384 reader (690 nm is subtracted from the 450 nm).

Reagents
Tartrazine (Sigma) solution are prepared in dimethyl sulfoxide (DMSO) at 3.0 g/L and dispensed at 1μL.

Qualification of the Bulk Reagent Dispenser
Qualification of the bulk reagent dispenser is done to confirm the accuracy of the volume dispensed. This bulk dispenser is qualified by dispensing volumes of water into a plate, which is then weighed on a balance, and the accuracy calculated. For our work, the Bulk Reagent Dispenser on the ALH (Caliper Life Sciences) was used.

Qualification of the Fixed-Cannula Array
Qualification of the 384 fixed array is done to confirm the accuracy of the volume dispensed. When testing the ALH for 1μL volume dispensing, 20μL water is added to the plates before or after (with chase) the tartrazine dispense. For our work, the 384 fixed-cannula array on the ALH (Caliper Life Sciences) was used.

Qualification Results
A gravimetric dispense test is done to determine the accuracy of the bulk (Table 1) and 384 fixed -cannula array (Table 2) dispense operations.

Fixed Cannula Precision Determination (1μL dispensing)

a. Water (20μL) was dispensed into each column of a 384-well plate with the Bulk Dispenser in a separate routine. Starting with a 3μL air gap, Tartrazine (5μL) was aspirated in a larger volume than needed (10μL). While still in the reservoir a 5μL volume was pushed out once. This leaves 5μL in the cannulas and, more importantly, residual on the outside of the cannulas. To control this the cannulas were touched off using a low volume dispense (0.2μL) on the surface of an upside down flat bottom plate followed by a small vertical tip touch. Finally the Tartrazine was consecutive dispensed (1μL) into each quadrant of a 384 well plate with the low volume head.

b. Starting with a 3μL air gap, a chase water volume of 20μL was aspirated followed by a 1μL air gap. Then tartrazine (1μL) was aspirated and finally the whole content of the cannulas was dispensed into a quadrant of a 384 well plate with the low volume head.

Precision Results
Absorbance was measured and the data exported to ExcelTM for CV determination. The results of the dispensed data for fixed cannula dispensing are shown below. The overall CV's that compromise every well of a 384 well plate are shown for each procedure tested. The average %CV per procedure is calculated in each table.

Discussion

In this study, we demonstrate the high level of precision and accuracy that can be expected of the Sciclone ALH Bulk Reagent Dispenser and the Low Volume Head with the 384 Fixed-Cannula Array. We achieved accuracy levels of 0.03 % for a volume of 50μL of water with the Bulk Dispenser. The dispense precision for the fixed cannulas were 0.52%, 0.22% and 3.07% for volumes of 50μL, 20μL and 1μL respectively. And accuracy levels of less than 1% for volumes of 50μL and 20μL with the low volume head and a 384 fixed cannula array.

The experimental data shows that excellent pipetting performance can be obtained from the Sciclone ALH (low volume head) equipped with the 384 Fixed-Cannula Array. The results surpass the published factory specifications ( CV <= 8%) at 1μL dispense volumes, likely as a result of good pipetting technique. We believe this % CV can even be reduced by using degassed water. We hadn't the opportunity to do so at the test facility.

We have found that residual on the outside of the cannulas becomes the most influential liquid when determining CV's in low volume dispensing. Optimized touch offs in combination with time delays at tip touches and very slow draw backs out of liquids increases remarkably the performance of the pipettor.



'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Multichannel Pipetting: How to Choose the Correct Pipettor
2. 96 Well Pipetting Precision and Accuracy Using the RapidPlate 96/384 Workstation
3. 384 Well Pipetting Precision and Accuracy Testing Using the Sciclone Advanced Liquid Handler
4. Sciclone ALH Bulk Dispense Precision Testing
5. Sciclone ALH 3000 Liquid Handler: Functional Characterization of the High Volume Head - Part 1: Plate-to-Plate Precision Testing
6. Precision Testing for the Sciclone Advanced Liquid Handler Z8 Option
7. Reliable Assessment of Micronuclei for Genotoxicity Testing in Mice or Rats
8. Restriction Endonucleases Undergo Rigorous QC Testing to Ensure Optimal Performance
9. Bio-Plex Cytokine Immunoassays and ELISA: Comparison of Two Methodologies in Testing Samples From Asthmatic and Healthy Children, Rev A
10. Testing Compressed Air Lines for Microbiological Contamination
11. Testing Compressed Air Lines for Microbiological Contamination
Post Your Comments:
(Date:1/15/2014)... 2014 TaiGen Biotechnology Company, Limited ("TaiGen") today announced ... a leading Russian pharmaceutical company, to develop and commercialize ... Federation , Turkey and ... a novel antibiotic for the treatment of bacterial infections ...
(Date:1/14/2014)... Histogen, Inc., a regenerative medicine company developing ... simulated embryonic conditions, today announced that they have entered ... for physician-dispensed aesthetic products containing Histogen’s proprietary multipotent cell ... an amendment to the existing license between Histogen and ...
(Date:1/14/2014)... York, NY (PRWEB) January 14, 2014 ... of Alliqua, Inc. (OTCQB: ALQA). Alliqua is an emerging ... to serve the wound care market. , Free report ... was restructured with a seasoned management team and Board, ...
(Date:1/14/2014)... 2014 iLab Solutions, the leader in core ... Director of Product Strategy. In this role, Michelle will work ... to guide in the development of iLab products. Her main ... possible benefit to the scientific community by offering the most ...
Breaking Biology Technology:TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 2TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 3TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 4Histogen and Suneva Medical Expand License for Cell Conditioned Media-based Aesthetic Products Internationally 2Histogen and Suneva Medical Expand License for Cell Conditioned Media-based Aesthetic Products Internationally 3EquitiesIQ Initiates Coverage of Alliqua, Inc. 2EquitiesIQ Initiates Coverage of Alliqua, Inc. 3iLab Solutions Announces Michelle Detwiler as the New Director of Product Strategy 2
... , EMERYVILLE, Calif., Aug. 6 Onyx Pharmaceuticals, ... to sell an aggregate of four million shares of its common stock ... million aggregate principal amount of its 4.00% convertible senior notes due 2016 ... options to purchase up to an additional 600,000 shares of its common ...
... , ALBANY, Ore., Aug. 6 Synthetech, Inc. (OTC ... of fiscal 2010, which ended June 30, 2009. Revenue for ... of $4.9 million in the first quarter of fiscal 2009. ... operating income of $314,000 for the same period last year. ...
... Md., Aug. 6 Vanda Pharmaceuticals Inc. (Vanda) (Nasdaq: ... of clinical-stage product candidates for central nervous system disorders, today announced ... on Monday, August 10, 2009, before the market opens. A full ... , , Mihael H. Polymeropoulos, M.D., President and ...
Cached Biology Technology:Onyx Pharmaceuticals Announces Pricing of Concurrent Offerings of Four Million Shares of Common Stock and $200 Million Principal Amount of Convertible Senior Notes 2Onyx Pharmaceuticals Announces Pricing of Concurrent Offerings of Four Million Shares of Common Stock and $200 Million Principal Amount of Convertible Senior Notes 3Synthetech Reports First Quarter Fiscal 2010 Results 2Synthetech Reports First Quarter Fiscal 2010 Results 3Synthetech Reports First Quarter Fiscal 2010 Results 4Synthetech Reports First Quarter Fiscal 2010 Results 5Synthetech Reports First Quarter Fiscal 2010 Results 6Vanda Pharmaceuticals to Announce Second Quarter 2009 Financial Results on August 10, 2009 2
(Date:4/24/2014)... treatment for a brain infection caused by fungus could now ... to study by University of Liverpool scientists. , Cryptococcus ... fatal but are relatively neglected in medical research. They ... Australasia and South East Asia and mainly affect people with ... a year. , The University research team has tested the ...
(Date:4/24/2014)... the largest river turtle in North America, weighing in ... century. Now researchers from Florida and the University of ... but three. , Examining museum specimens and wild ... ancient reptile. , Once heavily hunted for turtle meat ... Turtle Soup in the 1960s the riverine populations ...
(Date:4/23/2014)... chronic inflammatory disorders among people of lower socioeconomic status ... to the microbes that thrive in rural environments, according ... in the journal Clinical & Experimental Immunology , ... less access to green spaces may be more apt ... system dysfunction. , When our immune systems are working ...
Breaking Biology News(10 mins):Treatment for deadly yeast disease reduced to 3 days 2Two new US turtle species described 2Two new US turtle species described 3Rural microbes could boost city dwellers' health 2Rural microbes could boost city dwellers' health 3
... Mellon University,s Philip R. LeDuc and his collaborators in ... a protein that could ultimately unlock the mystery of ... role in the mechanics of biological processes in people. ... function of a protein that helps control cell behavior ...
... Reunion in the Indian Ocean and collaborating with researchers at ... sensitive night cameras to capture the first known occurrence of ... only is this the first time this behaviour has been ... who are better known for eating plants but ...
... number of scientists are merging methods and results from different ... team of researchers in a recent issue of Evolution ... new data, however, researchers who want to analyze previous findings ... the authors say. In a commentary piece in the journal ...
Cached Biology News:Carnegie Mellon's Philip LeDuc discovers new protein function 2First known instance of a cricket as an orchid pollinator captured on film by Kew scientist 2Biologists merge methods, results from different disciplines to find new meaning in old data 2Biologists merge methods, results from different disciplines to find new meaning in old data 3
Application: for RNase- decontamination , Registered Trademark of Ambion. ,Special grade: for molecular biology Density: 1.00 g/mL (20 C) Suitab tst: RNase decontamination in accordance...
With L-glutamine and potassium bicarbonate. Sterile, Insect cell culture tested...
... meet the needs of the ... medium supports fast cell growth ... while maintaining high cell viability ... using the Baculovirus Expression Vector ...
Mol wt: average mol wt12,360.96 Da by calculation...
Biology Products: