Preparation of Plates Using the Hamilton MicroLab Star

Introduction

The Hamilton MicroLab STAR is a robotic pipetting workstation. It is used for automated pipetting of liquid samples in medium to high throughput applications and has a high degree of flexibility.

The MicroLab STAR has a worksurface called a deck. The deck houses loadable carriers which hold reagent containers such as tubes, plates etc, and there are two tip waste containers. The pipetting arm has 8 pipetting channels which work independently. The CO-RE 96 Probe Head will pipette a block of 96 channels simultaneously. The carriers are automatically loaded onto the deck by the Autoload unit and there is a barcode reader for identification of plates, tubes etc. There is also a hand held bar-code reader. The instrument has a hinged front cover with magnetic switch.

Safety Notes

Read and sign the COSHH Risk Assessment and attached agreement, Task Risk Assessment and attached agreement and SOP and attached agreement before beginning the procedure.Labcoat, safety glasses and disposable nitrile gloves should be worn (gloves must be removed before using the computer). Do not obstruct the deck while the robot is in operation by, for example, placing your hands/head inside the front cover.The front cover must be closed in the correct position prior to commencing liquid transfer.Do not look into the laser beam of the barcode reader.Spillages should be cleaned up according to the cleaning procedure and reported to your supervisor or the safety officer.All faults with the robot must be reported immediately to your supervisor or the safety officer.

Materials

Hamilton MicroLab Star pipetting workstationTip Carrier : TIP_CAR_480_A00 (PN 182085/02)Plate Carrier : PLT_CAR_L5MD_A00 (PN 182365/01)Multiuse modular APE Carrier (PN 188039/00)CO-RE Low Volume Tips (10ul) without Filters (Hamilton)Tip Waste bagsAppropriate plates for the method that is to be run. The following plates are compatible: AB-0800-L, AB-1127, AB-1178, TF-0384, 4309849(ABI), AB-1111, 4306737(ABI).Plate spinning centrifuge → Sorvall Legend MACH 1.6

Procedure for turning on and initialising the MicroLab STAR

1. Turn on the Hamilton Microlab Star using the green switch on the front left hand side of the machine.

2. Log onto the computer and click on the ‘MicroLab STAR Run’ Desktop Icon (looks like a traffic light):

3. Go to ‘File’ on the top menu bar, click ‘Open’ and highlight the Method that is to be run. Refer to Appendix for details of the methods in use by the Nephrology Research Group. Click on ‘Open’ and the Method of choice will open up. Visually check that there is nothing obstructing the deck, and that the front window of the MicroLab STAR is closed.

4. Click on the button on the menu bar that looks like two cogs:

The control panel will open up.

5. Click on ‘Initialise Instrument’ and initialisation will proceed automatically. Initialisation needs to be performed before the first run of the day and following a break in activity eg after lunchtime.

6. Check the position of the barcode reader and if it is in the way, click on ‘Move Autoload’ to >45. The Barcode reader will move automatically to the right hand side of the deck. Click on ‘Close’.

7. *** Follow specific procedures described in liquid handling method of interest. ***

8. Ensure deck is left clear of all plastics and reagents and is ready for further use with all racks in position on the deck (i.e. not pulled out over the silver deck area).

Important Hints and Tips

If the robot fails to read a barcode, or if there is no barcode on your plate, and error message will be displayed. Click on ‘continue’ and then ‘execute’ to proceed with the method.

It is preferable to interrupt a program through the software by using the pause button on the screen, rather than opening the front window of the robot. Opening the front window will prevent the program from restarting at the current step.

If there is not enough DNA left in the stock plate for the robot to recognise, an error message will be displayed. Pick the option ‘ Bottom’ and click on ‘Execute’ to continue. This instructs the robot to go to the bottom of the wells.

**If an error occurs resulting in tips still being attached to the probes (8-channel or 96-core head), these must be removed before the program is continued. Dispose all tips to trash immediately → DO NOT aspirate as this may result in liquid in needles and severe damage to equipment **. Click on ‘Abort’ and you will be asked to check for horizontal collision. If clear, then click ’OK’. You will be asked if you want to abort to trash. Select ‘Yes’ or ‘OK’.

If a crash has occurred on the deck, a water test must be carried out before proceeding with any real samples in order to assess if any damage to the robot has occurred. To run a water test, rerun the method that you have been using with plates/tubes of water, coloured with a few drops of food dye (makes the aliquots easier to see). After completion of the method, check that the correct volumes have been dispensed.

The MicroLab STAR should be cleaned regularly. Refer to the protocol for ‘Cleaning the MicroLab STAR’. Cleaning is normally carried out between 3.00pm and 4.00pm on a Friday afternoon, and the Microlab STAR must not be used for 2 hours after cleaning is completed to allow for complete drying, so if you need to use the robot on Friday afternoon then arrange in advance with Jill Kilner.

In case of emergency and if AJ or Shirley are not available, contact Hamilton Robotics (local supplier Mason Technologies). Contact Name: Derek Tinsley, Analytical Team Manager for Mason Technology Ltd. Contact Numbers: Phone:  00353-1-4154476 Mobile: 00353-87-8071994, Fax: 00353-1-4154494