1. MALDI user guide
Contents
1. in Fig 6 to open this window Fig 7 Initially this window will be greyed out Check the SAMPLE PERIOD 1 in Fig 7 crs Page of 12 For REFLECTRON this should be 0 5ns and for LINEAR 5ns If the value is wrong click on the SECURITY tab 6 in Fig 6 type in the password and click APPLY then return to the ADC SETTINGS tab Click on the pull down menus for the SAMPLE PERIOD and select the correct value for the mode being used Instrument Settings X Flight Tube Inlet System Yacuum System Voltages Tuning Setup Security ADC Settings Signal Sensitivity 100 Trigger Threshold 250 um Input Signal Offset W Trigger Slope Positive Negative ee Sample Period 05 1 5 Fig 7 Click on the APPLY button then close the window The instrument is now ready to run samples Select the spot to analyse by typing the location into the SAMPLE box 1 in Fig 8 The PULSE 3 in Fig 8 DETECTOR 4 in Fig 8 SUPPRESSION 5 in Fig 8 and LASER 2 in Fig 8 are either set or adjusted as explained in the training The laser is steered using the cross hairs on the left 6 in Fig 8 which are also seen on the camera image on the right 1 in Fig 8a crs Page 8 of 12 The right hand PEAK DISPLAY screen shows each individual laser shot and the left hand PEAK DISPLAY screen shows the summed spectrum 10 shots summed Fig 9 0 0 200000 0 4002. 000 0 aw M sauna iL N ger 0128 Pulse 2050 Sample 51 41 Detector z2 70 Lock Mass I Suppression 500 H teady 6 Plate Carrier Loaded Pumped p Start B o m Documenti Microsoft WH MassLynx CRS CRS SPL MALDI micro MX c mas i Camera Control Fig 8 Bua al Fi Camera Control l m x Lucr a H IRefleckr Live Image Cursor 113 88 E Fig 8a crs Page 9 of 12 MALDI micro MX c asslynz crs pro acqudb crs ipr gm gd a 10 Shot Summed Individual Laser Spectrum Shot mo F jer po H perd E Ir WE Ea E I malga klp J Ca amp ie is a a cons SLILILILILI LI Fig 10 crs Page 10 of 12 a Tune Window Acquisition l X File Data File Mame TEST Browse Origin Mass Range Start Mass 500 a End Mass 3000 Laser 5 Laser Firing Rate Hz s rj 4 Shots per 5pectrum 0 rea Fig 10a To acquire data click on the ACQUIRE icon 1 in Fig 10 this will open a new window Click in the DATA FILE NAME box 1 in Fig 10a and type in a name for the data file bearing in mind the restrictions on using some characters If you wish to add a text description to the results printout type this into the TEXT box 2 in Fig 10a Type in the START MASS and END MASS values 3 in Fig 10a for the acquisition scan this does not have to be the same as used for the non storage acquisition PE
3. 3 select the INSTRUMENT SETTINGS menu 5 in Fig 3 to open the settings window Fig 5 crs Page 4 of 12 MALDI micro MX c masslyngscrs pro acqudbcrs ipr Pe a rm DA le a e Fig 3 Calibration pepmix cal cal Bm Ema AE Fig 4 crs Page 5 of 12 Instrument Settings X Flight Tube Inlet System Vacuum System Voltages Tunng Setup Security ADC Settings Flight Tube Flight Tube fei volts no Reflectron 5200 j olts et Negative 4node n Volts r a High Mass Detector Enable High Mase Detector Dynode fo L vers na Source 5 e 100 0 BOOOO0 1 eQO0000 1 0 0 200000 0 Suppression HOR H onae i JCO IE Fig 5 Check that the Flight Tube 1 in Fig 5 is at 5000 volts if not type this value in and press enter Click on OPERATE 2 in Fig 5 The RED square should turn GREEN and the red values in the Instrument Settings window will now show the voltages being applied Increase the FLIGHT TUBE voltage to its operating voltage of 12 000 volts in 2 000 volt increments press ENTER after each input and check that the voltage increases The REFLECTRON value should be at 5 200 volts and should not be changed If you have selected NEGATIVE ionisation the NEGATIVE ANODE slider 3 in Fig 5 will be active This should be raised from its starting voltage of O volts to its full operatin
4. AK DISPLAY The LASER FIRING RATE and SHOTS PER SPECTRUM 4 in Fig 10a should be 5 and 10 respectively as before To start the acquisition either press the ENTER key on the keyboard or the START button 5 in Fig 10a in the acquisition window crs Page 11 of 12 When you have finished running samples you should reset the FLIGHT TUBE in the instruments settings window to 5000 volts this can be done in 1 step and click the APPLY button Close the CHROMATOGRAM amp SPECTRUM windows turn off the CAMERA using the icon 2 in Fig 3 leave the instrument in OPERATE and remove your plate from the instrument Ensure the instrument has started the sample plate load sequence and the LOAD light 3 in Fig 1 is flashing before leaving REMEMBER TO TRANSFER YOUR DATA FILES FROM THE MALDI DATA SYSTEM TO ANOTHER STORAGE DEVICE FOR ARCHIVING crs Page 12 of 12
5. CRL MASS SPECTROMETRY FACILITY INSTRUMENT USER MANUAL MALDI MICRO Operating Instructions Basement Spec Lab 00 097 crs Page 1 of 12 This is a guide to using the MALDI Micro for those who have received training If you have any questions or problems whilst using this instrument please contact a member of the CRL Mass Spectrometry Facility Staff NOTE ALL USERS MUST OBTAIN TRAINING FROM A MEMBER OF THE MASS SPECTROMETRY STAFF BEFORE USING THE MALDI MASS SPECTROMETER Introduction The Waters MALDI micro MX can be operated in Linear or Reflectron modes Linear covers the whole mass range of the instrument 0 500K Daltons but has a low resolution approximately 1000 whereas Reflectron gives much higher resolution approximately 12 000 but the mass range is restricted 0 15K Daltons Whilst both Positive and Negative ionisation can be used most sample types are run in positive ionisation NOTE The use of matrices in sample preparation usually raises the effective starting mass from 0 to around 500 Daltons Sample Preparation The method used to prepare the sample and the specific matrix to use will depend upon the sample characteristics Chapter 6 pp83 102 in the Operators Manual which is kept by the instrument gives a description of the common matrices their preparation the type of compounds suitable for each matrix and preparation of standards Typically the sample should be at a concentration of 1 100pmol ul a
6. g voltage of 5 000 volts in 1 000 volt increments in the same manner as the flight tube voltage If you have selected LINEAR mode the REFLECTRON will be greyed out and the HIGH MASS DETECTOR will be active Click on the square ENABLE HIGH MASS crs Page 6 of 12 DETECTOR 4 in Fig 5 and increase the DYNODE voltage 5 in Fig 5 from 0 volts to its operating voltage of 5 000 volts in 1 000 volt increments as above Click the APPLY button 6 in Fig 5 for these changes to be set Click the TUNING SETUP tab 7 in Fig 5 to open the non storage acquisition tuning page Fig 6 Instrument Settings h X Flight Tube Inlet System Vacuum System Voltages Tuning Setup Security ADC Settings Mage range for tuning Start Mass End Mass 3500 Laser control for tuning Laser Firing Rate Hz 5 Shots per Spectrum 10 General instrument settings Manually set sample rate Low Mass Bias oe Fig 6 Type in values for the START MASS 1 in Fig 6 and END MASS 2 in Fig 6 to cover the mass range of your sample This range will be used for displaying the spectra on the screen when not acquiring data and can be different to the range used to acquire data The LASER FIRING RATE and SHOTS PER SPECTRUM 3 in Fig 6 should be set to 5 and 10 respectively Click on the APPLY button 4 in Fig 6 for the values to be set Click on the ADC SETTINGS tab 5
7. imately 2 minutes to start up and will bring up the main window Fig 2 if MassLynx is running then load your project as detailed below crs Page 3 of 12 BA MassLynx Ramirez New SPL File View Pum Help H F m Lal gt F ui Gi shortcut Th Queue fsp Status Spectrum Chromatogram Map Edit Samples e min m id ndn iii EL MS Method s oed M amp Tune 4 m EN 4 Eg 2 el e 2 Ci uw LL Fig 2 Load your project by clicking on the Open File icon 1 in Fig 2 and selecting your project Open the Spectrum and Chromatogram windows by clicking on the links 2 amp 3 in Fig 2 Open the TUNE page by clicking on the MS TUNE link 4 in Fig 2 This will bring up the tune page which is used to set up and run the instrument Fig 3 Click on the menu option CALIBRATION 1 in Fig 3 and select CALIBRATION ROUTINE This opens a calibration window Fig 4 Click on FILE 1 in Fig 4 check that it is looking in your project and open the calibration file for your run This will usually be called pepmix pos cal or bsa lin cal unless you are using a calibration file specific for your samples Close the calibration routine window Click on the CAMERA icon 2 in Fig 3 to turn on the sample imaging camera Select which mode you require by clicking either the LINEAR or REFLECTION icon 3 in Fig 3 Select which ionisation type you require by clicking on the or icon 4 in Fig
8. nd the matrix at 1mg ml Ideally both the sample and matrix should be prepared in the same solvent which should evaporate easily at room temperature The sample spots must be completely dry before the plate is put into the instrument to reduce the vacuum pump down time and the risk of arcing when the high voltage is applied to the extraction grid Instrument Operation To load a plate into the instrument press the UNLOAD button on the instrument 1 in Fig 1 When the flashing UNLOAD light changes to a steady LOAD light press the OPEN button 2 in Fig 1 to open the top cover this may take a bit of effort as there is still a small residual vacuum in the sample chamber Place the plate into the sample carrier platform ensuring that the corner indent is at the bottom right Ensure the top of the sample plate is level with the carrier platform before closing the lid as if itis raised it may damage the extraction grid Close the lid and press the LOAD button 3 in Fig 1 this will start the load sequence which will take approximately 1 2 minutes If the MassLynx software is not running wait for the flashing LOAD light to change to a solid UNLOAD light before starting the software as this will cause hardware software conflicts which will prevent the software working crs Page 2 of 12 Acs ei THIE SAN POSITION WIJHE FIG 1 If you need to start the MassLynx programme double click the MassLynx 4 1 icon on the desktop This will take approx
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