Navigation Links
Dispensing 0.5μL to "Dry" 384 Plates with the Sciclone ALH

Mike Benway, Larry Simonson and John Ung
Caliper Life Sciences Inc, Applications Laboratory, Hopkinton, Massachusetts, USA

Introduction

"Assay-Ready" plates prepared from compound or library stock plates in real time greatly accelerates the High Throughput Screening (HTS) process. The ability to transfer and dilute 0.5μL volumes is a prerequisite for this function. Here we present preliminary data on 0.5μL dry dispensing using the Caliper Life Sciences Sciclone Advanced Liquid Handler (ALH) Workstation, configured with a Low Volume Head and 384/1536 Fixed-Cannula Array.

Materials and Methods

Instrumentation

All testing was performed using a Sciclone ALH with a Low Volume Dispensing Head, and a Bulk Reagent Dispenser (Caliper Life Sciences). Absorbance was measured at 450nm using a Wallac Victor2 1420 Multilabel Counter (PerkinElmer).

Reagents

Tartrazine was obtained from Sigma Chemical Company, Catalogue number T-0388. A stock dispensing solution was prepared by dissolving 2.8g of tartrazine into 1L of DMSO. The stock solution was prepared for dispensing volumes of 0.5μL. Deionized water (DI) was used as a diluent after dispensing into the dry plate.

Accuracy Determination

Dispense accuracy is determined before measuring dry dispense precision. A calibration is performed to determine the volume dispensed. For the calibration, 50μL of deionized water is pipetted in to a 384 Well Flat Bottom, Untreated Polystyrene Assay Plates (COSTAR, Catalogue # 3702). Next, 0.5μL of the tartrazine solution is manually injected into each of 20 wells, successively, from a filled 10μL Hamilton micro syringe. The wells are manually mixed by multiple aspirate- dispense cycles using a 50μL hand pipettor. Any air bubbles are removed, and the plate is read for absorbance. The average absorbance reading for these manual standards is used in a single point calibration routine.

Sciclone Protocols

Here a single aspiration step is performed, followed by priming dispenses, then incremental dispenses into dry plates as follows:

Data Acquisition and Processing

Absorbance data is acquired on a Wallac Victor2 reader using the same read protocol as for the calibration plate. The raw results are exported to ExcelTM and %CV is calculated. The volume dispensed is calculated by comparison of signal to a manually prepared calibration standard.

Results

Testing has been done for three and five incremental plates using the method described above. For each run, the volume dispensed and %CV for each plate is calculated and summarized (Table 1). The results for the two runs are represented graphically (Figure 3).

Discussion

In this preliminary work we have found that the Sciclone ALH equipped with a Low Volume Head is capable of delivering 0.5μL of liquid into dry microplates with a precision of less than 6%CV. This is performed with no direct contact between the cannulas and the plate. In the above results, a "first plate" effect is observed, in which the first plate in a series has a higher volume. We believe this is due to residual tartrazine solution of the external surfaces of the cannulas that is removed as the first plate is dispensed.

Initial dry plate work was carried out using a 384 Cannula Array. The benefits of this array are that it can access both deep well plates and standard microplates. However, the larger cannulas do not permit formation of well formed droplets. Without such droplets, physical contact of the cannulas with the plates is required when transferring sub-microliter volumes to dry plates, potentially resulting in variable %CV's (unpublished data). For the above results, we used a shorter, finer 384/1536 Cannula Array designed for accessing 384 well plates or 1536 plates (by dispensing into four quadrants). This type of a cannula array can fully access standard microplates, or half height deep well plates. The main benefit of the smaller cannulas is that uniform droplets can form on the ends of the finer cannulas, allowing the dispense of the droplet onto the plate without contacting the plate.

The success of dry dispensing depends on assuring the Cannula Array and destination plates are parallel. Therefore, it is necessary to level any position in which sub-microliter dry dispensing is attempted. The deck of the Sciclone ALH is designed to be adjustable at each corner. The individual plate locators must also be level. It is also necessary to determine a precise dispense height for any non-contact dispense operation.

Physical factors are an important determinant of overall dispensing performance. Such factors include microplate composition, microplate surface treatments, solvent concentrations and compositions, solute (compound) characteristics and solute concentration. The current study represents 2.8g/L tartrazine dissolved with DMSO and dispensed into untreated, flat-bottomed, polystyrene plates only. Under these conditions we demonstrated that it is feasible to dispense sub microliter volumes to dry plates with very good precision.


'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Counting Colonies or Plaques Directly from Images of Agar Plates
2. Counting FlashPlates on the Wallac MicroBeta
3. Direct Counting of Millipore MultiScreen Filtration Plates.
4. Sciclone ALH Bulk Dispense Precision Testing
5. 96 Well Pipetting Precision Testing Using the Sciclone ALH with a 384 Fixed-Cannula Array
6. Sciclone ALH 3000 Liquid Handler: Functional Characterization of the High Volume Head - Part 1: Plate-to-Plate Precision Testing
7. Scalable Automation of the TempliPhi DNA Sequencing Template Amplification and DYEnamic ET Terminator Sequencing Kits on the Sciclone ALH 3000
8. Precision Testing for the Sciclone Advanced Liquid Handler Z8 Option
9. 384 Well Pipetting Precision and Accuracy Testing Using the Sciclone Advanced Liquid Handler
10. Automation of the Eppendorf Perfectprep BAC 96 DNA Kit on the Caliper Life Sciences Sciclone ALH 3000
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Dispensing Dry Plates with the Sciclone ALH

(Date:5/5/2015)... May 05, 2015 VetStem Biopharma, ... Animal Health, Jeffrey Schaffer, DVM, as Director of ... 15 years of experience in the veterinary regenerative ... addition to our veterinarian lead organization”. Dr. ... “Dr. Schaffer will be responsible for all technical ...
(Date:5/5/2015)... May 05, 2015 Research and Markets ... the addition of the "Separation Systems ... offering. Macromolecules which include protein, ... or are broken into smaller molecules, during ... separation requires various advanced systems like centrifugation, ...
(Date:5/5/2015)... TEL AVIV, Israel , May 5, 2015 ... that the Company was approved to continue trading ... companies. OTCQB Marketplace allows for ... an electronic broker-dealers. Investors can have real-time quotes ... The OTCQB Marketplace includes ...
(Date:5/5/2015)... , May 5, 2015  Renova™ Therapeutics, ... congestive heart failure and other chronic diseases, has ... independent analysis designed to reflect the healthcare patterns ... This model will be used to characterize the ... could have on those groups of patients and ...
Breaking Biology Technology:Leading Regenerative Veterinary Medicine Company, VetStem Biopharma announces the addition of Dr Jeffrey Schaffer as the Director of Veterinary Professional Services. 2Global Separation Systems Market in Biotechnology 2015-2019 - Trends and Forecasts for the $28 Billion Market 2Tikcro Technologies Announces Approval to Continue Trading on OTCQB 2RAND Health Advisory Services modeling heart failure patient outcomes to inform potential impact of Renova Therapeutics lead product candidate 2RAND Health Advisory Services modeling heart failure patient outcomes to inform potential impact of Renova Therapeutics lead product candidate 3
(Date:4/9/2015)... , April 9, 2015 Synaptics Inc. (NASDAQ: ... solutions, announced today that it will report financial results ... April 23, 2015, after the close of market. The ... and investors at 2:00 p.m. PT (5:00 p.m. ET), ... To participate on the live call, analysts and ...
(Date:4/2/2015)... 2, 2015 At its 2015 ACMG Annual ... , the American College of Medical Genetics and ... to its Board.  Members of the ACMG Board of ... forming and advancing its policies and programs. ACMG is ... "It,s an eventful time in medical genetics and ...
(Date:4/1/2015)... --  Medisafe ™, the leading global medication management platform ... and Android smartphones and tablets, announces today ... to allow patients to track and correlate their adherence ... will be able to visualize in real-time how taking ... as glucose levels and blood pressure. "Patients ...
Breaking Biology News(10 mins):Synaptics to Report Third Quarter Results on April 23 2American College of Medical Genetics and Genomics Announces New Board Members: Dr. Louanne Hudgins is ACMG President-Elect 2American College of Medical Genetics and Genomics Announces New Board Members: Dr. Louanne Hudgins is ACMG President-Elect 3American College of Medical Genetics and Genomics Announces New Board Members: Dr. Louanne Hudgins is ACMG President-Elect 4American College of Medical Genetics and Genomics Announces New Board Members: Dr. Louanne Hudgins is ACMG President-Elect 5American College of Medical Genetics and Genomics Announces New Board Members: Dr. Louanne Hudgins is ACMG President-Elect 6American College of Medical Genetics and Genomics Announces New Board Members: Dr. Louanne Hudgins is ACMG President-Elect 7American College of Medical Genetics and Genomics Announces New Board Members: Dr. Louanne Hudgins is ACMG President-Elect 8Medisafe Announces Launch of Biometric Integration, Enabling Patients to Stay Adherent to Their Meds and See Tangible Health Results 2
... is a delicacy costing up to SEK 8,000 a kilo ... has discovered that this tasty fungus also grows wild in Sweden. ... Sweden, and definitely in Japan once these discoveries become known there," ... is a family of many different species of fungi. One ...
... available in <A HREF=",http://chinese.eurekalert.org/zh/emb_releases/2010-06/du-wmi062410.php,">Chinese . DURHAM, ... far greater than those found in seawater, it,s the ... up posing a more serious health threat to humans ... University researchers, is in the seawater itself. The ...
... Beijing, June 27th, 2010 The Elsevier Foundation, ... twelve talented women scientists from Africa, the Middle East, ... excellence. The announcement was made at the Third World ... and International Conference, Women Scientists in a Changing World, ...
Cached Biology News:Japanese gourmet mushroom found in Sweden 2Japanese gourmet mushroom found in Sweden 3Why mercury is more dangerous in oceans 212 women scientists announced as winners of Elsevier Foundation TWOWS Awards 2
... S2 cells are used for heterologous protein ... (DES). The S2 cell line was derived ... hours old) Drosophila melanogaster embryos (1). ... room temperature without CO2 and is easily adapted ...
... syringe pump series combines Harvard Apparatuss best pump ... a wide range of configuration options. A lookup ... of syringes from all the major manufacturers. A ... the pertinent information from across the lab. All ...
... For our novel expression vectors, we in-troduced ... dual controls: (i) at tran-scriptional level at the ... replication level by amplifying the plasmid copy number, ... is that it retains practically all of the ...
... This CLS number is a new ... product number. If showing no availability yet, ... (C6421) or contact customer service for assistance. ... Comp Dim: surface area 21 cm ...
Biology Products: