Navigation Links
96 Well Pipetting Precision and Accuracy Using the RapidPlate 96/384 Workstation

Lynn Jordan1 and Brent Butler2
1Caliper Life Sciences Inc., Applications Laboratory, Hopkinton, Massachusetts, USA
2GlaxoSmithKline, Department of Molecular Screening, Research Triangle Park, North Carolina, USA


Initially, work in 96 well microplates was commonly carried out using manual pipetting. As the demand for greater throughput of plate processing has grown, automated liquid handling has become a necessity for most laboratories. The most critical aspect of an automated liquid handler is to understand its performance, typically determined by precision and accuracy.

At Caliper Life Sciences, precision and accuracy testing is achieved by dispensing a tartrazine solution into microplates containing water. This is referred to as a "liquid-to-liquid" dispense. The purpose of this paper is to report the performance obtained by dispensing 0.5 to 2μL aqueous samples containing tartrazine into 96 well plates containing water. Precision and accuracy results are shown for this work.

Materials and Methods

Testing was performed using a RS-232 RapidPlate 96/384 Workstation (Caliper Life Sciences) using 100μL Automation Certified Pipette TipsTM (Caliper Life Sciences). A Bulk Dispenser (Caliper Life Sciences) was utilized for bulk dispensing across the columns of the microplate (secondary dispensing). Absorbance was measured at 450nm using a Wallac Victor2 Multilabel Counter (PerkinElmer).

Tartrazine (Sigma Chemical Company, Catalogue number T-0388) is dissolved in deionized water. Concentrations of 6.0, 1.4, 0.7 and 0.14g/L corresponding to both 1 and 2, 5, 10 and 25μL dispenses respectively are prepared by dissolving the tartrazine into 1L of deionized water. The tartrazine solution was dispensed into plates containing a volume of degassed deionized water. The deionized water was degassed by bubbling helium through the water for a period of 15 minutes.

Qualification of a Reagent Dispenser
Qualification of a bulk reagent dispenser is done to confirm the accuracy of the volume dispensed. When testing the RapidPlate for low volume dispensing, 100μL of degassed water is added to the plates both before and after the tartrazine dispense. This is done on an independent dispenser. 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, a Bulk Reagent Dispenser on a Sciclone Liquid Handler (Caliper Life Sciences) was used.

Calibration of the RapidPlate
The RapidPlate is calibrated for accuracy prior to precision testing (see, Jordan L and Cunningham J "Calibration of Dispensing Accuracy in the RapidPlate, Application Note, Caliper Life Sciences Inc., in press 2001). A gravimetric confirmation of the dispensed volumes is repeated to verify the accuracy of the calibrated dispensed volumes.

Setting up the RapidPlate
In the RapidPlate software the "Procedure Editor" is selected. Next, using the "Setup Tab" the deck layout is created with a reservoir, a tip box and one White 96 Well Clear-bottom un-treated plate (COSTAR), See Figure 1. The reservoir is filled with the tartrazine dispensing solution appropriate for the volume to be tested. The level of the liquid in the reservoir should be such that the tips just break the surface of the solution. The 96 Well plate contains 100μL of degassed water, dispensed by a qualified liquid handler. A clean box of 100μL pipette tips are used for every dispense.

Running the RapidPlate Procedure
For the Liquid-to-liquid dispense a RapidPlate procedure was written to attach tips, aspirate a volume of tartrazine solution from the reservoir, and then dispense into a clear bottom plate containing 100μL of degassed water. The procedure was designed to dispense into a single plate. Apart from appropriate changes in volumes, the same protocol was used for all volumes tested (Figure 2).

Mixing the Plates
After pipetting the 5 plates, 100μL of deionized water was added to each plate using a qualified reagent dispenser to aid in the mixing of liquids in the well. Next, the plates were read.

Reading the Microplates
Plates were read for absorbance at 450nm. The raw results were exported to ExcelTM and CV's were calculated for each volume.


A gravimetric dispense test is done to determine the accuracy of the dispense operation after calibration of dispensing accuracy (Table 1).


The experimental data shows that excellent pipetting performance can be obtained from the RapidPlate 96/384 Workstation. After calibration, we achieved accuracy levels of less than 1% error for volumes of 50, 20, 10, and 5μL, and errors of 2% and 30% for volumes of 2μL and 1μL respectively. While 30% error on the 1μL accuracy may give reason for concern, this can be easily reduced by entering additional gravimetric data points at these lower volumes when performing the calibration.

The dispense specifications for precision of the RapidPlate 96/384 Workstation are less than 5%CV for volumes of 1- 10μL, and 2% for volumes greater than 10μL, (for a liquid to liquid aqueous dispense). In our study, the CV's achieved are better than the specifications, especially taking into account actual volumes dispensed at 1 μL were significantly less, as the calibration was performed across the entire dispensing range, and not optimized for the lower volumes.



Page: All 1 2 3 4

Related biology technology :

1. Multichannel Pipetting: How to Choose the Correct Pipettor
2. 96 Well Pipetting Precision Testing Using the Sciclone ALH with a 384 Fixed-Cannula Array
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. Combine High Yield, Great Accuracy and the Prevention of Carry-over Contamination by Using the Novel Expand High FidelityPLUS PCR System
8. Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
9. Oil-Free PCR Using the Hot Top Assembly
10. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
11. Signal Transduction Reporting Systems Using Cis-Acting Enhancer Elements
Post Your Comments:

(Date:6/23/2016)... CAMBRIDGE, Mass. , June 23, 2016 /PRNewswire/ ... the development of novel compounds designed to target ... compound, napabucasin, has been granted Orphan Drug Designation ... in the treatment of gastric cancer, including gastroesophageal ... cancer stemness inhibitor designed to inhibit cancer stemness ...
(Date:6/23/2016)... Prostate Cancer Foundation (PCF) is pleased to announce 24 new Young ... cancer. Members of the Class of 2016 were selected from a pool of ... More About the Class of 2016 PCF Young Investigators ... ... ...
(Date:6/23/2016)... Durham, NC (PRWEB) , ... June 23, 2016 , ... ... Odense University Hospital in Denmark detail how a patient who developed lymphedema after being ... (fat) tissue. The results could change the paradigm for dealing with this debilitating, frequent ...
(Date:6/23/2016)... ... June 23, 2016 , ... ClinCapture, the only free validated ... will showcase its product’s latest features from June 26 to June 30, 2016 ... on Disrupting Clinical Trials in The Cloud during the conference. DIA (Drug ...
Breaking Biology Technology:
(Date:4/15/2016)...  A new partnership announced today will help ... in a fraction of the time it takes ... life insurance policies to consumers without requiring inconvenient ... Diagnostics, rapid testing (A1C, Cotinine and HIV) and ... weight, pulse, BMI, and activity data) available at ...
(Date:4/14/2016)... Israel , April 14, 2016 ... Authentication and Malware Detection, today announced the appointment of ... assumed the new role. Goldwerger,s leadership appointment ... on the heels of the deployment of its platform ... BioCatch,s behavioral biometric technology, which discerns unique cognitive and ...
(Date:3/31/2016)... 2016   LegacyXChange, ... "Company") LegacyXChange is excited to release its ... to be launched online site for trading 100% guaranteed ... will also provide potential shareholders a sense of the ... an industry that is notorious for fraud. The video ...
Breaking Biology News(10 mins):