Addendum to PAL System User Manual PAL MALDI Option
Contents
1. Lower Tool retainer nut o gt gt a E PEEK nut o gt Spotting probe Figure 8 Assembling the MALDI Tool 2 5 Figure 9 2 6 Figure 10 Addendum to PAL System User Manual PAL MALDI Option Installation Variations of the MALDI Tool The PAL MALDI Option can be configured in various different modes Depending on the application the plumbing method may be slightly different Some examples of possible configu rations are shown below 1 2 3 Installation variations of the MALDI Tool PEEK tubing no matrix addition HPLC column inside HPLC system Fused silica tubing no matrix addition HPLC column inside HPLC system Fused silica tubing online matrix addition via Nano Y connector HPLC column inside MALDI Tool Installation of MALDI Plate Adapters Insert the appropriate MALDI plate carefully to its corresponding MALDI plate adapter Depending on the plate type some adapters are equipped with location pins For accurate positioning all Target adapters are spring loaded on their right side Insert the adapter into a PAL Stack Drawer or a PAL MT Trayholder as shown in Fig 10 page 18 Double check if the MALDI plates and its adapters are firmly in place If regular 96 384 well microplates for micro fraction collection are used the plates are inserted directly without adapter into a PAL Stack Drawer or a PAL MT Trayholder Installation of MALDI Plate Adapters note 3 3 12. 25 2 6 3 1 3 2 55 w Addendum to PAL System User Manual PAL MALDI Option PAL MALDI Option Operating Manual How to use this manual General System Overview Specifications Hardware requirements Software requirements Installation Unpacking the Components Installing the PAL MALDI Tool Parkstation Installing the Magnetic Gripper Basic Installation of the MALDI Tool Installation Variations of the MALDI Tool Installation of MALDI Plate Adapters PAL MALDI Option Object Position Adjusting Object Reference Positions Adjusting Position of MALDI Tool Parkstation Setting MALDI Tool Offset PAL MALDI Option Utility Functions Cycle Composer Control Appendices Definition of Terms Naming Conventions PAL MALDI Option Spare Parts 10 11 13 13 14 14 14 16 21 21 22 23 Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 List of Figures PAL MALDI Option General System Overview PAL MALDI Option Parts Attaching the Tool Parkstation to the X axis Attachment of Mounting Claws Location of the Tool Parkstation Attaching the Magnetic Gripper to the PAL Z axis Installing the Tool Parkstation Installing the MALDI Tool Installation variations of the MALDI Tool Installation of MALDI Plate Adapters Object Position of MALDI Tool Parkstation MALDI Tool Setup Object Screen Utility Menu Screen Object Tools
3. 3 2 Figure 11 Addendum to PAL System User Manual PAL MALDI Option PAL MALDI Option Object Positions Adjusting Object Reference Positions Remove the Syringe Adapter from the Injection Unit before performing the follow ing steps It is assumed that the necessary firmware version and the PAL MALDI Option object list is installed on the PAL instrument All existing PAL Objects e g PAL Sample Stacks Trayholders etc must already be properly defined according to the corresponding HTS or HTC PAL Operation Manual The objective is to adjust the reference positions for the PAL MALDI Option Make sure the MALDI Tool Parkstation including the MALDI Tool and tubing the Magnetic Gripper the MALDI Plate adapter including the appropriate target plate or microplate are properly mounted to the PAL Adjusting Position of MALDI Tool Parkstation The reference position of the MALDI Tool Parkstation is shown in Fig 11 The bottom side of the Magnetic Gripper should slightly touch the top side of the blue Parkstation tube Position of MALDI Tool Parkstation Addendum to PAL System User Manual PAL MALDI Option 1 Remove the MALDI Tool from Tool Parkstation 2 When the Job Queue menu is displayed complete the following sequence Menu Setup Objects Tools MALDISpt 3 After selecting MALDISpt the X Y Z positions for the MALDI Tool Parkstation ParkingPos will be displayed see Figure 12 ParkingPo
4. Object Position of MALDI Tool Offset Cycle Composer example macros Addendum to PAL System User Manual PAL MALDI Option note Addendum to PAL System User Manual PAL MALDI Option How to Use this Manual The manual is divided into two sections PAL MALDI Option Operating Instructions Appendices The PAL MALDI Option Operating instructions are intended for frequent PAL users or new users that are experienced in using automated systems to perform existing analytical methods The HTS PAL or HTC PAL must be installed and set up properly before the PAL MALDI Spotter Operating Instructions can be used The Appendices provide information about the PAL MALDI Options and Spare Parts Addendum to PAL System User Manual PAL MALDI Option PAL MALDI Option Operating Instructions 1 General System Overview Figure 1 PAL MALDI Option General System Overview The PAL MALDI Option micro collector spotter is an add on PAL module for micro scale fraction collection or MALDI Spotting together with any Capillary LC or Nanoflow LC system The collec tion of very low volume fractions requires low delay volumes and high position accuracy of the capillary tip The principles of operation for the PAL MALDI Option micro collector spotter will be illustrated in this manual along with descriptions on how to set up the hardware and change the configuration This enables the user to run the instrument with optimal performance 1 2 13
5. Check Clear Movto Home Pos Pos Zero Utility Menu Screen Object Tools Scroll to ToolOffset Z and set the value to 20 0 mm This step is important if the Tool Tip has been changed Otherwise if the new tip is longer than before it will hit the plate when moving to the Tray in the next step Press F1 Check Offs The Injection Unit will open the Stack drawer if present pick up the MALDI Tool and move to the Tray selected in TeachPoint sample position 1 Addendum to PAL System User Manual PAL MALDI Option Figure 14 Object Position of MALDI Tool Offset 7 Select ToolOffset Z and move down until the Tool Tip is close to the MALDI Target surface Because the polished MALDI Target plates act as a mirror the correct position is reached when the tip and its mirror image are close together In case of a microplate adjust the Z value to the top surface of the plate 8 Adjust ToolOffset X and Y so that the Tool Tip is perfectly centered to the sample 1 position 9 Press F1 Rest Tool The MALDI Tool will be restored to the Parkstation and eventually the stack drawer will be closed note The adjustment of the MALDI Tool Offset must be done carefully Small variations result in improper drop deposition Addendum to PAL System User Manual PAL MALDI Option 5 Cycle Composer Control note _ __ To operate the PAL MALDI Option via Cycle Composer software it is
6. Specifications Part Number Flow Rate Min time slice Delay volume Fraction vessel capacity MALDI Target capacity Supported Targets Cooling option Transfer tubing kit Software Control PAL Firmware Wetted parts Net Weight Addendum to PAL System User Manual PAL MALDI Option PAL MALDIOpt max 300ul min 3 seconds per fraction or target 20 fractions or spots min approx 3ul with 1 meter PEEK Tubing ID 65um approx 10nl if column is installed inside MALDI Tool HTS PAL max 23 Shallow plates 11 Deepwell plates 594 2ml vials HTC PAL max 11 Shallow plates 5 Deepwell plates 270 2ml vials 96 or 384 well compatible up to 24 depending on vendor and PAL instrument type Targets of Bruker Daltonics ABI Sciex Waters Agilent Optional thermostatted Fraction and Target storage 4 C 40 C 2m PEEK tubing ID 65um OD 1 16 1 pc PEEK Nano Y connector for 360um capillary tubing 1 pc PEEK Union 10 32 to 10 32 0 005 thru hole 10 pcs FEP Sleeves for 340um 380um capillary tubing 3 pcs Coated Spotting Tips By Cycle Composer Software Version 1 5 3 or higher or PAL ICC Version 3 0 or higher PEEK Fused Silica 550 g Hardware requirements The PAL MALDI Option can be used with any HTS PAL or HTC PAL equipped with a side slitted Injection unit PAL LC can not be used with PAL MALDI Option Software requirements The PAL MALDI Option requires PAL Firmware 3 0 or higher It can be
7. and Collect to MALDI Inject sample selected in Tray and Vial inthe Air Volume Plate sample list Pre Clean with Solvent 1 Start spotting to MALDI plate after Collection Pre Clean with Solvent 2 Start Delay measured from injection Cleaning Pre Clean with Sample of the syringe and injector must be terminated Filing Speed before Filing Strokes The MALDI plate is selected with Tray2 in the nject to sample list njection Speed Pre Inject Delay Post Inject Delay Post Clean with Solvent 1 Post Clean with Solvent 2 Valve Clean with Solvent Valve Clean with Solvent Collection Start Position Collection Start Delay Collection Time umber of Collections Inject and Collect to MALDI Inject sample selected in Tray and Vial inthe Air Volume Plate with Waste Start sample list Pre Clean with Solvent 1 Start spotting to MALDI plate after Collection Pre Clean with Solvent 2 Start Delay measured from injection Cleaning Pre Clean with Sample of the syringe and injector must be terminated Filing Speed before Filing Strokes The MALDI plate is selected with Tray2 in the nject to sample list njection Speed Before collection starts the probe is moved to Pre Inject Delay Collection Waste Position so the first spot will Post Inject Delay be accurate Post Clean with Solvent 1 Post Clean with Solvent 2 Valve Clean with Solvent 1 Valve Clean with Solvent 2 Collection Start Position Collection Start Delay Collec
8. controlled by Cycle Composer software 1 5 3 or higher or any data system that controls the PAL using PAL ICC e g Analyst ChemStation Empower EZChrom MassLynx Xcalibur 2 1 Figure 2 Swe OnaAunFWN gt Addendum to PAL System User Manual PAL MALDI Option Installation Unpacking the Components The MALDI Option is shipped in one single box Check for the completeness for following items some parts are preassembled Ipc PAL MALDI Tool Ipc PAL MALDI Tool Parkstation 1pc Magnetic Gripper incl 2 pcs mounting screws 1 5m PEEK tubing ID 65um OD 1 16 Ipc PEEK Nano Y connector for 360um capillary tubing Ipc PEEK Union 10 32 to 10 32 thru hole 0 13mm 10pcs FEP Sleeves for 340um 380um capillary tubing 3pcs Coated Spotting Tips 1pc Parkstation Waste Tube 10pcs PEEK nuts 10 32 1 16 Z i i z a a a 10 i i Y 7 ja py F i s gt d Lt a gt D 9 SSS 2 SS Gp SS Up K SS Wer S EE Za Z Z a PAL MALDI Option parts Addendum to PAL System User Manual PAL MALDI Option 2 2 Installing the PAL MALDI Tool Parkstation Before beginning the installing process determine approximately where the PAL MALDI Tool Parkstation should be placed It must be positioned near the left end of the PAL X axis 1 I
9. naming conventions for MALDI Targets and Microplates Following these conventions will allow PAL MALDI Setup s to be pre configured for certain applications will simplify software backups and application development and will improve technical support and training Tray Type Vendor P N Tray Description MT96 various Standard 96 position shallow microplates DW96 various Deep well 96 position microplates MT384 various High density 384 position shallow microplates VT54 CTC 54 position 2ml vial tray BRK16x24 Bruker 384 spots MTP Anchor Chip steel ABI10x10 ABI Sciex P N V700664 100 spots SS numbered with etched circles MM 8x12 Waters P N M880675CD1 96 spots MM 16x24 Waters P N M880356BD1 384 spots MM 8x12 Waters P N 186002322 96 spots MassPREP DIOS target AGI 8x12 Agilent P N G1972 60025 96 spots note ___ The list above reflects the status of August 2005 Check the delivered CD ROM or with your CTC Analytics representative for new MALDI Target Types 22 Addendum to PAL System User Manual PAL MALDI Option C PAL MALDI Option Spare Parts Part no Description MZ 24 01 Magnetic Gripper incl screws MM 010 01 PAL MALDI Tool Parkstation P 773 PEEK Nano Y connector P 779 01 PEEK Union thru hole 0 13mm PAL TubeWaste Parkstation Waste Tube PAL MaldiTip 3 3pcs Spotting Tips with special coating 1560 15m PEEK Tubing ID 65um OD 1 16 F 331NX 10pcs PEE
10. Addendum to PAL System User Manual PAL MALDI Option Addendum to PAL System User Manual PAL MALDI Option Printing History Edition 1 September 2005 CTC Analytics AG reserves the right to make improvements and or changes to the product described at any time without notice CTC Analytics AG makes no warranty of any kind with regards to this product including but not limited to the implied warranties or merchantability and suitability for a particular purpose In no case shall CTC Analytics AG be liable for any coincidence ore consequential damages in connection with or arising from the use of this document 2005 CTC Analytics AG All rights reserved Neither this publication nor any part of this publi cation may be copied photocopied reproduced translated or reduced to any electronic medium or machine readable form without the prior written permission of CTC Analytics AG except as allowed under the copyright laws CTC Analytics AG acknowledges all trade names and trademarks used as the property of their respective owners General Considerations Other Hazards Addendum to PAL System User Manual PAL MALDI Option Safety Information A Changes or modifications to this unit not expressly approved by the party responsible for compliance could void the user s authority to operate the equipment The user shall be made aware that if the equipment is used in a manner not specified by the manufacturer the protection prov
11. K Nuts 10 32 1 16 F 242X 10pcs FEP Sleeves for 340um 380um capillary tubing 23
12. anual PAL MALDI Option Attaching the Magnetic Gripper to the PAL Z axis Basic Installation of the MALDI Tool The MALDI Tool is already preassembled using the PEEK tubing ID 0 0025 OD 1 16 the PEEK Union 10 32 to 10 32 the PEEK nuts 10 32 1 16 and one coated spotting tip according Fig 8 view parts on Fig 2 page 13 Put the MALDI Tool into the Tool Parkstation Guide the end of the PEEK tubing through the the red fitting mounted at the rear side of the Tool Parkstation Make sure the tubing length allows the MALDI Tool to reach the whole working area but keep it as short as possible If the necessary tubing length is properly adjusted tighten the red fitting on the Tool Parkstation view Fig 7 page 17 Connect the PEEK tubing to your HPLC system Keep the length as short as possible to minimize delay volume The MALDI Tool is pre assembled to show its correct installation It is delivered with 1 5m PEEK tubing ID 0 0025 OD 1 16 For other ID s contact your PEEK or Fused Silica tubing supplier Note that improper MALDI Tool connections can lead to leaks or the formation of dead delay volume both may result in poor performance Addendum to PAL System User Manual PAL MALDI Option to HPLC Figure 7 Installing the Tool Parkstation PEEK Tubing to Tool Parkstation and HPLC a Upper Tool retainer nut a MALDI Tool PEEK nut D Tool Spacer PEEK Union
13. assumed that the user is familiar with the PAL Cycle Composer control software The PAL MALDI Option can only be used together with the PAL control software Cycle Com poser Make sure the hard and software requirements meet with your installation before operating the MALDI Option see page 12 1 2 Hardware requirements 1 3 Software requirements Every PAL MALDI Option is shipped with a CD ROM containing 5 different MALDI application examples see Fig 16 page 26 These examples may be used to get familiar with the MALDI Option and are a good starting point to develop your own applications Copy these examples to your Cycle Composer method folder Addendum to PAL System User Manual PAL MALDI Option Macro Name Macro Description Macro Variables Collect to MALDI Plate Start spotting to MALDI plate after Collection Collection Start Delay Start Delay Collection Time The MALDI plate is selected with Tray and Number of Collections starting position with Vial from the sample list Collect to MALDI plate with Start spotting to MALDI plate after Collection Collection Start Delay Waste Start Start Delay Collection Time The MALDI plate is selected with Tray and Number of Collections starting position with Vial from the sample list Collection Waste Position Before collection starts the probe is moved to Collection Waste Position so the first spot will be accurate Inject
14. ided by the equipment my be impaired When you use the PAL System follow the generally accepted procedures for quality control and methods development When you use the PAL System in the field of chromatographic analysis and you observe a change in the retention of a particular compound in the resolution between two compounds or in peak shape immediately determine the reason for the changes Until you determine the cause of a change do not rely on the separation results A To avoid injury during PAL System operation keep your hands away from the syringe A Do not operate the PAL System without the safety shield The safety shield must be installed for safe operation A To avoid injury observe safe laboratory practice when you handle solvents change tubing or operate the PAL System Know the physical and chemical properties of the solvents you use See the Material Safety Sheets from the manufacturer for the solvents in use A Use caution when working with any polymer tubing under pressure e Always wear eye protection when near pressurized polymer tubing e Do not use polymer tubing that has been severely stressed or kinked e Do not use polymer tubing in particular not PEEK or Tefzel tubing with Tetrahydrofuran THF Dimethylsulfoxid DMSO chlorinated organic solvents concentrated mineral acids such as Nitric Phosphoric or Sulfuric acids or any related compounds to above listings 1 1 1 2 1 3 2 1 22 23 24
15. nj A Method defines how the samples are processed The elements of a Method are a Cycle a Syringe and a Parameter List Methods have names with up to 8 characters and can be edited copied and deleted Method Parameters are associated with the Cycle operations User assigned Parameter values define how a processing operation is performed A zero Parameter value will disable a Cycle operation Cycle parameters are application specific Objects are data structures describing the properties of physical modules Certain modules e g a Stack require several objects A Trayholder can hold one or more trays Each Trayholder has a reference position X Y Z co ordinates that defines its location A Stack is a particular type of Trayholder that is designed to hold micro plates A six drawer Stack holds 12 standard micro plates two in each drawer A three drawer Stack holds six deep well micro plates two in each drawer A Tray that holds multiple samples Trays are defined by designating the Tray Type see below and the Trayholder Tray names are used to Identify the sample source within a PAL Job A Tray Type defines the pattern and sampling sequence of sample locations within a Tray Standard Tray Types include shallow and deep well 96 position Microplates and Trays that hold 1 2 10 and 20ml vials or MALDI Targets 21 B Naming Conventions Addendum to PAL System User Manual PAL MALDI Option This section recommends standard
16. nstall the Tool Parkstation with the mounting clamp teeth fitting into the grooves on the bottom of the X axis The Parkstation must be mounted near the left end of the PAL X axis Be sure that the clamp fits completely into the grooves Tighten the Torx screw until the mounting clamp is firmly in place h W Figure 3 Attaching the Tool Parkstation to the PAL X axis 2 Double check if the Tool Parkstation mounting clamp is correctly attached to the PAL X axis see Figure 4 TN CN o o Wrong Correct Figure 4 Attaching PAL objects to the PAL X axis Connect one end of the delivered Waste tube to the Tool Parkstation waste port Insert the other end into an appropriate waste container note The distance between the right edge of the Tool Parkstation and the left edge of a PAL Stack PAL Stack Cooler or PAL Trayholder must be gt 85mm see Figure 5 Addendum to PAL System User Manual PAL MALDI Option Figure 5 Location of the Tool Parkstation 23 Installing the Magnetic Gripper 1 Turn OFF the PAL Power Supply 2 Slide down manually the black syringe carrier inside the Z Crossrail until the two threaded holes located on its left side are visible 3 Locate the two supplied Torx screws and attach the Magnetic Gripper securely to the black syringe carrier see Figure 6 Figure 6 24 note Addendum to PAL System User M
17. otter tip should just be above the center of position 1 of the MALDI Target or the center of the first microplate well note ___ The procedure of adjusting the Tool Offset has to be repeated every time a change is made at the MALDI Tool assembly especially if the Spotting Tip is exchanged note note Figure 13 5 Addendum to PAL System User Manual PAL MALDI Option PAL MALDI Option Utility Function The PAL MALDI Option Utility function selectable from the Menu screen provides quick access to MALDI positioning parameters that may need to be fine adjusted The MALDI Tool adjustment applies for all Trays there is no individual adjustment for each Tray To have a consistent precision from plate to plate it is important that the Trays have been fine adjusted using the Tray utility function See the PAL manual for a description of the Tray utility function Insert the MALDI Tool into the Tool Parkstation Insert the MALDI Target Adapter incl Target or the microplate into the Sample Stack Drawer or Trayholder close all Stack Drawers Switch on the instrument Starting with function key F1 complete the following sequence Menu Utilities Tool MALDISpt For TeachPoint select the Tray that holds the MALDI Target or microplate inserted in step 2 This Tray will be used to adjust the MALDI Tool position see Figure 15 page 23 MALDISpt TeachPoint ToolOffset X ToolOffset Y ToolOffset Z 2005 07 12
18. s X ParkingPos Y ParkingPos Z TeachPoint ToolOffset X ToolO0ffset Y ToolOffset Z 2005 07 12 1321625 Check Clear Movto Home Pos Pos Zero Figure 12 MALDI Tool Setup Object Screen 4 Select the item ParkingPos Y with the cursor bars and press ENTER The Injection Unit will move to the actual defined Y axis position 5 Do not change the value because it is preset to an approximate value 6 Press the inner knob to enter the ParkingPos Y value 7 Repeat steps 4 6 first for ParkingPos Z and last for ParkingPos X The X position is preset to 0 and has to be adjusted 8 Fine tune the X Y and Z values until the black Magnetic Gripper fits exactly to its position shown in Fig 11 9 Verify the defined X Y Z Positions by pressing F1 Check Pos note __ Pressing the F3 button Movto Zero will move the injection unit to the Zero Position Addendum to PAL System User Manual PAL MALDI Option 33 Setting MALDI Tool Offset The offset between injection syringe needle and coated spotter tip has to be adjusted after installation of the MALDI Option to ensure the required precision for MALDI spotting After the hardware and software has been set up properly follow the procedure described in Chapter 4 PAL MALDI Option Utility Function to adjust the tool offset Reference position of the MALDI Tool Offset offset between Injection syringe needle and coated spotter tip is described in Fig 13 The sp
19. tion Time Number of Collections Collection Waste Position Micro Fraction Collection Start collecting to plate after Collection Start Collection Start Delay Delay Collection Time The plate is selected with Tray and starting Number of Collections position with Vial from the sample list Well Offset Well Offset is half the diameter of the collection well Well penetration is taken from the plate Figure 15 Cycle Composer MALDI Example Macros 20 A Job Queue Job Cycle Method Method Parameters Object Trayholder Stack Tray Tray Type Addendum to PAL System User Manual PAL MALDI Option Appendices Definition of Terms A Job Queue is a list of sample processing Jobs Jobs are executed in the order displayed on the JOB QUEUE menu screens New Jobs may be added to the queue while samples are being processed A Job contains the information needed by the PAL to process multiple samples by the same processing steps The elements of a Job are a Method and a Tray that defines the location of the samples to be processed For identification Jobs are automatically numbered from 01 to 99 and then restarting with 01 when they are added to the Job Queue A Cycle consists of the specific operations necessary to process one sample The Cycle operations are repeated for each sample within a Job Cycles are designed for specific applications The Standard HTS PAL Cycle is named LC I
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