1. Expert Training Technical Recourses picoSpin NMR Spectrometers Dean Antic, Ph.D. Senior Applications Scientist

2. Exercise 1 Out of Box Setup Exercise 2 Sample InjectionSample Exercise 3 autoShim Script Exercise 4 Measuring SNR Exercise 5 onePulse Script Exercise 6 Data Processing Exercise 7 Troubleshooting Exercise 8* Cartridge Exchange Exercise 9* Cartridge-change Shimming Documentation Kits & Manuals Calculators SNR & Tx Training Videos Exercise Schedule *Time permitting

3. Exercise 1: Out of Box Setup 1.Unpack you picoSpin; 2.Make power connections; 3.Turn on the pS-45 unit; 4.Connect the Ethernet cable; 5.Make network adapter modifications; 6.Using Firefox, browse to the pS-45; 7.Browse to Temperature page; 8.Adjust set-point to 42 ° C* and submit; 9.Let the pS-45 warm up. Today we will be controlling the picoSpin using a direct/static connection to the computer. * The pS-45 is shipped with a set-point temperature of 42 ° C RJ-45 Ethernet Connector RS-232 Port Power Connection +12 V: 110-240 VAC ON/OFF Switch

4. Network Adapter Settings for Direct Connection Windows 7 Navigate to the networking configuration: Select Control Panel → Network and Internet → Network and Sharing Center Select Change adapter settings Select Local Area Connection Select Internet Protocol Version 4 (TCP/IPv4) Click Properties Select Alternate Configuration Select User configured Type in the following fields: IP address: 192.168.42.1 Subnet mask: 255.255.255.0 Click OK, Click Close Windows XP Navigate to the networking configuration: Select Control Panel → Network Connections Select Local Area Connection Click Properties Select Internet Protocol Version (TCP/IP) Click Properties Select Alternate Configuration Select User configured Type in the following fields: IP address: 192.168.42.1 Subnet mask: 255.255.255.0 Click OK, Click Close, Click Close *Other Operating Systems

5. picoSpin-45 spectrometer including Pre-fitted capillary cartridge 12 VDC modular power supply Power cord Ethernet cable Starter packet Disposable syringe and needle Port plugs Inlet and outlet port adapters Inline filter Contact details 12 month licence to Mnova Quick start guide Factory test report What’s In The Box

6. Documentation Documentation available in HTML and PDF format at http://www.picospin.com/support Available documents Software revision: 0.8.0 (current release) Software revision: 0.8.1 (for 1/16” & 1/32” fittings) User Manual QuickStart guide FAQ Accessory Kits (for 1/16” & 1/32” fittings) pS-45 Classroom Accessory Kit pS-45 Laboratory Accessory Kit pS-45 Cartridge Maintenance Kit for 1/32” fittings only

7. Factory Test Report T EST R EPORT Date: 12-19-11 Operator: J. Price picoSpin-45 serial number: 11120004 Cartridge serial number: 11121009 Software version: app 0.80.0-11 build 0.8.0-33 Shim settings: attached page FID plot: attached page SNR report: attached page The Test Report provides factory default settings, serial numbers, shim values and images of FIDs verifying and certifying the shipped picoSpin-45 spectrometer as having meet factory specifications. Attached Documents Shimmed FID Shims SNR

8. Web Browser Interface There are four main pages of the interface Run Files Temperature System And two menu buttons scripts shims Accessing the web browser user interface (UI) Using Firefox point your web browser at the unit's IP address The IP address is shown on the front-panel LCD display at power-up

9. System Page Preferences Run Name Enables saving of named runs; Expert Mode Controls the starting condition of the show more/show less button in scripts; Display Plots at Right Controls position of the data plots wrt to parameters list; Settings Set the default IP address when DCHP is not available (OS reboot required); Set Date & Time (used for labeling saved runs); Set Time Zone Version Currently installed software and firmware versions; Restart Restart the web server CPU; System Update Update system software and web browser interface; Use only when update file and installation instructions are received.

10. Temperature Page The Temperature page UI provides access to the control interface to magnet temperature controller Plot Features Displays latest 6 hours of data Data refresh every 30 seconds Automatic Y-axis re-scaling Plot types Magnet temperature plot Magnet temperature sensor data displayed in Celsius ( ° C) Magnetization dependence on magnet temperature Magnetization Ambient/case temperature Room temperature + heated dissipated from inner-enclosure electronics Heater output +10 volts; 16-bit, 0 to 65535 (2 16 ) full scale Controller button Magnet set-point temperature – maximum setting 45/60 ° C (high temperature unit). Adjustable PID controls Heater On/Off switch Closed loop On/Off switch Controller settings are not changed until the *Submit* button is pushed. From Ambient, 30 – 120 mins is required to reach stabilized magnet set-point

11. Files Page The Files page UI provides access to data, settings and scripts stored on the instrument for download and reuse Run Data onePulse experimental parameter and spectrum archive autoShim parameter archive Individual file download and named, compressed archiving Shims Saved shim files Scripts Factory default experiment scripts

12. Run Page The Run page UI provides access to pulse sequence experiment scripts onePulse autoShim search Experiment scripts are selected via the scripts button and values are adjusted in the parameter fields according to script and experiment  onePulse script fields  1.Execute script 2.Expert/basic mode 3.Parameter fields

13. Exercise 2: Sample Injection Samples are introduced into the capillary cartridge by injection picoSpin-45 spectrometers are shipped with a Starter Kit containing 1-1 mL disposable polypropylene syringe 1 22 gauge blunt tip dispensing needle 2 PEEK plugs 1 Syringe Port 1 Inline Filter 1 Disposable 0.22 µm syringe filter *Classroom accessory kits replicate the starter kit & include syringe filters Starter Kit – 1/32” Fitting Starter Kit – 1/16” Fitting

14. Exercise 2: Sample Injection – Starter Kit Attaching the drain tube Insert the tip of the drain tube assembly into outlet fitting Finger tighten PEEK nut by applying gentle pressure Place the end of drain tube into small waste container Injecting a sample Using a polypropylene syringe Draw up 100 mL of the sample solution (400 mL if using syringe filter) Orient tip upwards, gently tapping syringe to dislodge any bubbles Attach syringe filter (if used) to the tip of the syringe Attach fill tube assembly to syringe/syringe filter Insert tip of fill tube into inlet fitting Finger tighten PEEK nut by applying gentle pressure Gently press the syringe plunger to inject sample Monitor drain tube to ensure residual bubbles have been purged After injection replace fill and drain tubes with PEEK plugs

15. Exercise 2: Sample Injection – Lab Kit Syringe Attaching the drain tube Insert the tip of the drain tube assembly into outlet fitting Finger tighten PEEK nut by applying gentle pressure Place the end of drain tube into small waste container Injecting a sample Using a glass gastight syringe Draw up 40 µL of the sample solution Draw solution slowly to avoid bubble formation Insert needle into inlet fitting Finger tighten PEEK nut by applying gentle pressure Gently press the syringe plunger to inject sample Monitor drain tube to ensure residual bubbles have been purged After injection replace fill and drain tubes with PEEK plugs

16. Exercise 2: Sample Injection Inject water for shimming

17. Exercise 3: The autoShim Script Magnet’s Field Z Shim Field Z BoBo Z Shimming is a process of optimizing the magnetic field homogeneity over the sample volume of the RF coil. Shimming is accomplished by passing small DC currents through coils. The picoSpin-45 has 9 shim coils: 3 for generating linear gradients 5 for generating quadratically varying fields 1 for generating a uniform field The autoShim script uses a search algorithm to find optimal shim settings.

18. Shimming Q&A When to check the shim quality of the picoSpin 45? When instrument is first turned on and after temperature stabilization; When instrument is power cycled for more than a few minutes; When the magnet set-point temperature is changed; Once daily (during normal operational use). Are there different types of shim procedures? Yes, there is Maintenance shimming; Set-up shimming; Cartridge-change shimming; Shimming from scratch. Are the shim procedures the same? No, some shim procedures are complex and take longer. How often should one shim the picoSpin-45? Maintenance shimming should be performed daily. Do I need to shim after changing the sample? No. Loss of shim will not occur upon changing samples. What sample requirements is there for shimming? Shimming is performed on a water; Acetone and cyclohexane can also be used. autoShim Script Parameters **Detailed Parameter Fields Description

19. ParameterValue tx frequencyproton Larmor frequency (MHz) max iteration120 test run linear increments0.005 quadratic increments0.02 target rms1 pulse width40 us or best 90-deg time acq. points1000 zero filling8000 rx recovery delay500 us T1 recovery delay8 s bandwidth4 kHz post-filter atten.10 phase correction0 degrees (or any value) exp. filter0 Hz max plot points400 max time to plot250 ms min freq. to plot-1000 Hz max freq. to plot+1000 Hz magnitude Shimming Shimming is a critical step in maintaining the picoSpin-45 in top operational form This table shows typical autoShim script parameters Good shims yield High resolution spectra High signal sensitivity Bad shims yield Low signal resolution Reduced signal sensitivity Parameters table are provided for Maintenance shimming; Maintenance shimming QuickStart (Set-up) shimming; QuickStart (Set-up) shimming Cartridge-change shimming; Cartridge-change shimming Shimming from scratch. Shimming from scratch Today we will perform QuickStart and Cartridge-change shimming

20. ParameterValue tx frequencyproton Larmor frequency (MHz) max iteration120 test run linear increments0.005 quadratic increments0.02 target rms1 pulse width40 us or best 90-deg time acq. points1000 zero filling8000 rx recovery delay500 us T1 recovery delay8 s bandwidth64 kHz post-filter atten.6 phase correction0 degrees (or any value) exp. filter0 Hz max plot points400 max time to plot12 ms min freq. to plot-32000 Hz max freq. to plot+32000 Hz Magnitude QuickStart Shimming: 1 Discover your signal using Zoom Search Step 1 1.Browse to the autoShim script 2.Enter a Run Name 3.Check/adjust the following parameters 4.Run the script 5.Position your signal to 100-300 Hz Moving the Tx frequency 1.44.93702 MHz 2.9 – 100 KHz 3.3 – 10 kHz 4.7 – 1 kHz 5.0 – 100 Hz 6.2 – 10 Hz 6.If signal is at -1200 Hz then 1.Add -1200 Hz to Tx frequency 2.44.93582 7.If the signal is at 1200 Hz then 1.Add +1200 Hz to Tx frequency 2.44.93822 8.Run the script again

21. QuickStart Shimming: 2 ParameterValue tx frequencyproton Larmor frequency (MHz) max iteration120 test runUNCHECKED linear increments0.005 quadratic increments0.02 target rms1 pulse width40 us or best 90-deg time acq. points1000 zero filling8000 rx recovery delay500 us T1 recovery delay8 s bandwidth4 kHz post-filter atten.10 phase correction0 degrees (or any value) exp. filter0 Hz max plot points400 max time to plot250 ms min freq. to plot-800 Hz max freq. to plot+800 Hz Magnitude Step 2 1.Enter/change Run Name 2.Adjust the parameters in RED 3.Run the script 4.When completed, test run and execute the script once again This will take 20-40 mins Some Terminology 1.Iteration – one complete Simplex loop 2.Cycle – one pulse within an iteration 3.Increment – step size On average the algorithm will make 2.4 cycles/iteration. Do the math At T1 = 8s, 1000 pts and 2.4 cycle/iter. (8s + 0.250s)*2.4*120 = 39.6 min

22. Maintenance Shimming ParameterValue tx frequencyproton Larmor frequency (MHz) max iteration120 test run linear increments0.005 quadratic increments0 target rms1 pulse width40 us or best 90-deg time acq. points1000 zero filling8000 rx recovery delay500 us T1 recovery delay8 s bandwidth4 kHz post-filter atten.10 phase correction0 degrees (or any value) exp. filter0 Hz max plot points400 max time to plot250 ms min freq. to plot-800 Hz max freq. to plot+800 Hz magnitude Maintenance shimming is performed once daily (when instrument is in use) Maintenance shimming is similar to Step 2 of QuickStart shimming except adjusting the quadratic shims isn’t always necessary (but advisable) Step 1 1.Enter a Run Name 2.Browse to the autoShim script 3.Check/adjust the following parameters 4.Run the script 5.Position your signal to 100-300 Hz 6.Run the script again Other shim procedures

23. Shim Do’s, Don’ts and Evaluation The quality of shim is evaluated against Test report FID and spectrum signal Test report spectrum peak height values User FID and spectrum signal User spectrum peak height values DO Do keep your instrument temperature stabilized 24/7 Do check the shim daily when in use Periodically when not in use Do perform Maintenance shimming daily when in use Do include quadratic increments for in maintenance shimming Do save named shim files Do include date and magnet temp in name Do capture a screen shot of shimmed FID/spectrum DO NOT Do not use less than 100 iterations when shimming Do not shim when the magnet temperature is fluctuating Do not shim using more than 1000 pts Other tips A successful shim will write values to the UI – SAVED THEM You must execute the autoShim script with test run to write values to shim electronics An aborted shim WILL NOT write values to the UI autoShim has made improvements

24. Exercise 4: Measuring SNR – Using the search Script peak height baseline RMS noise Signal-to-noise ratio (SNR) is a measure of the sensitivity of an instrument With the picoSpin we can measure SNR using the search script

25. Calculating Theoretical SNR 600 = manufacturing SNR spec (wrt H 2 O) LW = Signal line weight 1/2 = 2 water protons M i = sample molarity 1/55 = molarity of water N = number of scans Click for SNR Calculators

26. ParameterValue start frequencyproton Larmor frequency (MHz) stop frequencyproton Larmor frequency (MHz) frequency step-10 kHz SNR10 noise window start-800 Hz noise window end-400 Hz pulse width40 us or best 90-deg time acq. Points4000 rx recovery delay500 us T1 recovery delay8 s bandwidth4 kHz post-filter atten.10 phase correction0 degrees (or any value) exp. filter0.5 Hz max plot points4000 max time to plot1000 ms min freq. to plot-100 Hz max freq. to plot+800 Hz live plot Magnitude Search Script 1.Navigate to the search script 2.Check/adjust the following parameters 3.Shift your Tx frequency so that it appears at +600 Hz 4. *Hint* you can read the signal position in the text window below 5.Once your signal is positioned wait 30 sec and run the script 6.Make a note of your SNR

27. Exercise 5: The onePulse Script The onePulse script is used to perform pulse experiments for acquiring NMR spectra The main features of this script are Saving named experiments Acquiring multiple scans (pulses) Spectral alignment Saving averaged data Saving individual scans Inject a sample

28. ParameterValue tx frequencyproton Larmor frequency (MHz) scans1 pulse width40 us or best 90-deg time acq. points3000 rx recovery delay500 us T1 recovery delay8 s bandwidth4 kHz post-filter atten.10 phase correction0 degrees (or any value) exp. filter0 Hz max plot points400 max time to plot250 ms min freq. to plot-500 Hz max freq. to plot+500 Hz zero filling8192 align-avg. data live plot JCAMP avg. JCAMP ind.Unchecked The onePulse Script Step 1 1.Browse to the onePulse script 2.Enter a Run Name 3.Check/adjust the following parameters 4.Run the script 5.Position your signal with the right-most peak near 100 Hz 6.Run the script again 7.Make adjustments as needed 8.Apply phase correction Clockwise/counterclockwise rotation 9.Set Scans = 16 10. Execute onePulse script

29. Exercise 6: Data Processing Spectra are processed and presented using Mnova picoSpin data is stored in JCAMP-DX format Navigate to ‘Files’ page Select experiment Select “fid.avg.jdx” file to download and open in Mnova or Select ‘Download All Data’ button to tar and gzip all data Process data in Mnova or preferred NMR analysis package Data processing Open data file Baseline correction Zero fill (+ LP Forward to 32K) Phase correct (zero and first order) Apodize: exponential (<1.0 Hz) and Sine-bell (5-20 °) Reference

30. Data Processing – Opening file 1.Navigate to data storage directory 1.Right click and choose ‘open with’ 2.Select ‘MestReNova’ 3.Check the option ‘Always use the selected program to open this kind of file’ 4.Or double-click if file association exists

31. Data Processing – Mnova Basics Main page

32. Data Processing – Mnova Basics Spectrum View toolbar NMR Process toolbar Zoom In/Out Full Spectrum Fit to Height Main toolbars Apodization Fourier Transform Phase correction Base Line Correction Reference Cursor *The small triangle indicates a drop-down menu

33. Data Processing – Mnova Basics Step 1: Baseline Correction Clicking this icon will allow automatic baseline correction Step 2: Phase correction Select Manual Correction under the Phase Correction icon and a window will appear Click Biggest to change center of gravity, or use scroll bar to manually adjust COG PH0: hold left-mouse button and scroll PH1: hold right-mouse button and scroll Hold Ctrl key for fine tuning

34. Data Processing – Mnova Basics Step 3: Zero Filling Under the FT icon select Zero Filling. Use the following settings: 32K points Select LP Filling Check Forward LP Step 4: Apodization Under the Apodization icon use the following settings: Exponential: 1 Hz Sine-Bell: 5-20 degrees First Point: 0.5 Click OK Click Fit to Height icon

35. Data Processing – Mnova Basics Step 5: Reference aligning Click the TMS icon A red vertical cursor will appear Move cursor to a peak for aligning and click A window will appear Under the Solvents button there is a list of common solvent shifts In New Shift input a value Click OK Rescale as necessary Save your work

36. Documentation Classroom Kit Laboratory Kit Classroom Kit Laboratory Kit Maintenance Kit Field Service Inlet Fitting 1/16” fittings 1/32” fittings Manuals and FAQs 1/16” fittings1/32” fittings Accessory Kits Software rev. 0.8.1 User Manual QuickStart Guide FAQ Software rev. 0.8.1 & 0.8.0 (legacy) User Manual QuickStart Guide FAQ 0.8.0 UM and FAQ available

37. Calculators Tx Calculator SNR Calculators

38. Training Videos ACDLabs Basic autoShim - part 1 autoShim - part 2 Mnova Basic - Apodization - part 5 Mnova Basic – DL File - part 1 Mnova Basic - PC_BLC - part 3 Mnova Basic - TMS - part 4 Mnova Basic - ZF - part 2 Mnova-8 Real-FT fix Path Sample Injection Zoom Search picoSpin YouTube Channel Portable MP4’s

39. MOL Structure Resources Websites Chemspider.com Acros.be Chemicalbook.com Search for.mol structures Programs ACDLabs/Chemsketch Free academic only License Chemdoodle

40. MoMo Temperature ( ° C ) TcTc Magnetization-temperature curve Magnetization is inversely dependent on magnet temperature Magnetization is inversely dependent on magnet temperature The Larmor slope is ~ -40 kHz/ ° C (~700 ppm/ ° C) The Larmor slope is ~ -40 kHz/ ° C (~700 ppm/ ° C) For each ±1 ° C the Tx frequency must be adjusted ±40 kHz For each ±1 ° C the Tx frequency must be adjusted ±40 kHz Magnetization-Temperature Curve

41. Tx Larmor Frequency/Temperature Calculator Click for Tx Calculator

42. Exercise 7: TroubleShooting The most common troubleshooting service calls arise a lack of understanding of the Tx frequency, and misinterpretation of the FID and spectrum plots. There are typically three reasons underlying a troubleshooting call 1.Air in in the RF coil 2.Poor shim 3.Misplaced Tx frequency Below are some typical troubleshooting scenarios 1.Good FID, good spectrumGood FID, good spectrum 2.Poor FIDPoor FID 3.No FID, good spectrumNo FID, good spectrum 4.Good FID, no signalGood FID, no signal 5.No FID, no spectrumNo FID, no spectrum How do we evaluate the problem? What do we offer as a solution?

43. Capillary Cartridge Sub-panel Flange

44. Exercise 8: Cartridge Exchange Cartridge Maintenance is important aspect of maintaining a picoSpin-45 Pre-filter sample or use a syringe filter Displace samples with clean solvents Avoid precipitation reactions (important for dissolved solid sampling) Displace samples using appropriate solvents Park water, ethanol or acetone for overnight storage Clean cartridges with 60-100 µL injections of EtOH, acetone, CHCl 3 or other compatible solvents Replace frit-filter on inlet fitting (outlet fitting does not have a frit-filter) Replace frit-filter In extreme cases where damage has occurred to the cartridge it will have to be replaced Following a instructions for replacing the cartridge and re-shimming the magnet

45. Cartridge Exchange Step 1 Turn off and unplug the unit 1.Remove knurled sub- panel screws 2.Using a long-handled 3/32 in hex wrench loosen the cartridge flange screws 3.Lower and rest sub- panel on front panel  

46. Cartridge Exchange Step 2 1.Attach the cartridge removal to the cartridge flange 

47. Cartridge-change Shimming Step 3 1.Holding the tool in one hand and sub-panel in the other gently remove the cartridge from the spectrometer Step 4 1.Reinstall new cartridge 2.Switch on the unit and temperature stabilize the magnet before shimming 

48. ParameterValue tx frequencyproton Larmor frequency (MHz) max iteration200 test runChecked linear increments0.05 quadratic increments0 target rms1 pulse width40 us or best 90-deg time acq. Points1000 zero filling8000 rx recovery delay500 us T1 recovery delay8 s bandwidth64 kHz post-filter atten.6 phase correction0 degrees (or any value) exp. Filter0 Hz max plot points400 max time to plot12 ms min freq. to plot-32000 Hz max freq. to plot+32000 Hz magnitudeChecked Cartridge-Change Shimming:1 Cartridge shimming requires additional steps Step 1 1.Browse to the autoShim script 2.Check/adjust the parameters in Red 3.Run the script 4.Position your signal within ±500 Hz 5.Run the script again 6.Uncheck test run and execute script 7.When finished move on to Step 2

49. ParameterValue tx frequencyproton Larmor frequency (MHz) max iteration100 test runChecked linear increments0.05 quadratic increments0 target rms1 pulse width40 us or best 90-deg time acq. Points200 zero filling1000 rx recovery delay500 us T1 recovery delay8 s bandwidth16 kHz post-filter atten.8 phase correction0 degrees (or any value) exp. filter0 Hz max plot points400 max time to plot12 ms min freq. to plot-8000 Hz max freq. to plot+8000 Hz magnitudeChecked Cartridge-Change Shimming:2 Step 2 1.Check/adjust the parameters in Red 2.Run the script 3.Position your signal within to ±500 Hz 4.Run the script again 5.Uncheck test run and execute script 6.Move on to Step 3

50. ParameterValue tx frequencyproton Larmor frequency (MHz) max iteration100 test runChecked linear increments0.01 quadratic increments0.1 target rms1 pulse width40 us or best 90-deg time acq. Points200 zero filling1000 rx recovery delay500 us T1 recovery delay8 s bandwidth4 kHz post-filter atten.10 phase correction0 degrees (or any value) exp. filter0 Hz max plot points400 max time to plot50 ms min freq. to plot-2000 Hz max freq. to plot+2000 Hz magnitudechecked Cartridge-Change Shimming:3 Step 3 1.Check/adjust the parameters in Red 2.Run the script 3.Position your signal within to ±500 Hz 4.Run the script again 5.Uncheck test run and execute script 6.Move on to Step 4

51. ParameterValue tx frequencyproton Larmor frequency (MHz) max iteration100 test runChecked linear increments0.005 quadratic increments0.02 target rms1 pulse width40 us or best 90-deg time acq. Points1000 zero filling8000 rx recovery delay500 us T1 recovery delay8 s bandwidth4 kHz post-filter atten.10 phase correction0 degrees (or any value) exp. Filter0 Hz max plot points400 max time to plot250 ms min freq. to plot-2000 Hz max freq. to plot+2000 Hz magnitudechecked Cartridge-Change Shimming:4 Step 4 1.Check/adjust the parameters in Red 2.Run the script 3.Position your signal within to ±500 Hz 4.Run the script again 5.Uncheck test run and execute script Your instrument is now shimmed!

52. ParameterValue start frequency46.0 Hz stop frequency43.5 Hz frequency step-8 kHz SNR20 noise window start-4000 Hz noise window stop-1000 Hz pulse width40 us or best 90-deg time acq. Points200 zero filling8000 rx recovery delay500 us T1 recovery delay8 s bandwidth16 kHz post-filter atten.8 phase correction0 degrees (or any value) exp. Filter0 Hz max plot points400 max time to plot12 ms min freq. to plot-8000 Hz max freq. to plot+8000 Hz magnitudechecked Shimming From Scratch Shimming from scratch is an extremely rare event and occurs only on the production floor at picospin in Boulder, CO. All shims & test reports lost, phone lines, the internet and picoSpin’s servers would have to simultaneously fail for one to be placed in a shimming from scratch position. In any event, these steps are required 1.Browse to the search script 2.Check/adjust the parameters 3.Run the script 4.Locate signal 5.Browse to autoShim script 6.Follow Cartridge-change Shimming procedure

53. Troubleshooting

54. Computer Trouble Cannot establish a connection to the Instrument Is the instrument physically connected to the network or LAN? Is your network adapter properly configured?

55. Sample Handling Trouble High back-pressure Blocked Filter Back flush the filter with a clean solvent Replace the in-line filter assembly Complete loss of signal Immediately after injection Injected a bubble, re-inject sample During acquisition Sample/solvent is too volatile for the set point temperature Lower set point temp. and reshim Use an alternative solvent Loss of signal strength over time Inject shim solvent and run autoShim script. Compare to historical values

56. Good FID Good spectrum

57. No FID? Good spectrum, but at 0 Hz No FID?

58. Poor FID Poor spectrum

59. Good FID, No signal? Good FID No spectrum Good cost function

60. No FID? ?

61. Appendix Images

62. Test Report: Shims

63. Test Report: Shimmed FID

64. Test Report: SNR

65. Network Adapter Settings for Direct Connection Cont…. Windows Vista Navigate to the networking configuration: Select Control Panel → Network and Internet Select Control Panel → Network and Internet Select Network and Sharing Center Select Network and Sharing Center Select Unidentified Network Select Unidentified Network Click View status Click View status Select Properties Select Properties Select Internet Protocol Version 4 Select Internet Protocol Version 4 Click Properties Click Properties Select Alternate Configuration Select Alternate Configuration Select User configured Select User configured Type in the following fields: IP address: 192.168.42.1 IP address: 192.168.42.1 Subnet mask: 255.255.255.0 Subnet mask: 255.255.255.0 Click OK, Click OK Click OK, Click OK Linux Debian In Debian edit the file /etc/networking/interfaces Modify eth0 section as follows: Modify eth0 section as follows: iface eth0 inet static address 192.168.42.1 address 192.168.42.1 netmask 255.255.255.0 netmask 255.255.255.0 network 192.168.42.0 network 192.168.42.0 broadcast 192.168.42.255 broadcast 192.168.42.255 gateway 192.168.42.1 gateway 192.168.42.1

66. Internet Explorer Configuaration The picoSpin-45 supports Internet Explorer (IE) versions 8 and 9. Please ensure the following settings are enabled. Turn off caching Go to Internet Options! General tab Under Browsing history click Settings Enable the radio button Every time I visit the webpage (beneath Check for newer versions of stored pages:) Change script settings Go to Internet Options! Advanced tab Under Browsing check Disable script debugging (Internet Explorer) Uncheck Display a notification about every script error

67. Scripts Button Factory Scripts called from the Scripts button on the Run page Scripts contain last run parameters Scripts contain last run parameters These are not the same as Factory Default scripts on the Files pages These are not the same as Factory Default scripts on the Files pages Scripts are programs which carry out experiment instructions by writing pulse sequence and shim values to the NMR and shim electronics autoShim performs searches autoShim performs searches onePulse is the experiment script for acquiring data onePulse is the experiment script for acquiring data search is a Tx frequency search script and performs signal-to-noise ratio (SNR) search is a Tx frequency search script and performs signal-to-noise ratio (SNR)

68. Shims Button 1.Shim settings can be named and saved to a file for future referralShim settings can be named and saved to a file for future referral 2.Magnet temperature is for reference onlyMagnet temperature is for reference only 3.Linear gradient, quadratic and uniform field shim current valuesLinear gradient, quadratic and uniform field shim current values x, y, z zx, zy, xy, z 2, x 2 -y 2 zx, zy, xy, z 2, x 2 -y 2 B 0 B 0 Values are scaled from 0-1 and not actual currents   The Shims button provides UI access to Manually adjust shim settings Save shim values

69. System Page: Preferences Changing these settings requires a system Restart Changing these settings requires a system Restart

70. System Page: Settings Changing the Default IP address requires a rebooting the DCHP server Changing the Default IP address requires a rebooting the DCHP server Changing system clock does not require a system reboot Changing system clock does not require a system reboot

71. System Page: Version As new software revisions become available, communicating system version information to Tech Support is essential for troubleshooting As new software revisions become available, communicating system version information to Tech Support is essential for troubleshooting System update to latest revision will be required for Tech Support System update to latest revision will be required for Tech Support

72. System Page: Restart Restarting the web server CPU does not interrupt power to temperature controller Restarting the web server CPU does not interrupt power to temperature controller

73. System Page: Update System software is proprietary Updates contain unit serial number dependent public/private key encryption Tech support will require system registration and latest firmware installation Successful update require ~5 mins; Where systems “hang”, power cycling is required **All Scripts Shims, and data files will be overwritten during the update process

74. Files Page: Run Data     1.Clickable Run DataClickable Run Data 2.Last Run = Run DataLast Run = Run Data 3.Run Name & Script infoRun Name & Script info 4.Reload parameters optionReload parameters option 5.Script parameter fieldsScript parameter fields 6.Clickable download/Delete run optionClickable download/Delete run option 7.Download archived Run Data optionDownload archived Run Data option  1/3

75. Files Page: Run Data Download or open FID file 1.Click fid-avg.jdx file linkClick fid-avg.jdx file link 2.Browse to NMR softwareBrowse to NMR software 3.Or save to computerOr save to computer 4.Click OKClick OK Deleting a saved Run 5.Click “Delete this Run”Click “Delete this Run” 6.Click “Yes” or “NO”Click “Yes” or “NO” *Off load data regularly     2/3

76. Files Page: Run Data Download experiment archive 1.Click “Download All Data” buttonClick “Download All Data” button 2.A ‘.tar’ file archive will be automatically downloadedA ‘.tar’ file archive will be automatically downloaded Extract using Windows zip facility, 7-Zi, Winrar, etc…   3/3

77. Files Page: Shims      1.Clickable Current and Default shim filesClickable Current and Default shim files 2.User saved shim filesUser saved shim files 3.QuickStart factory default shim filesQuickStart factory default shim files 4.Reload shims optionReload shims option 5.Saved shim valuesSaved shim values 6.Delete shim file optionDelete shim file option 7.Download shim file optionDownload shim file option

78. Files Page: Scripts Factory Scripts Default factory settings These scripts are never overwritten Software revisions replace these files Settings are not specific to magnet or cartridge Use only when all other scripts are lost

79. Temperature Controller    1.PID controller tuning parameter (optimized factory settings)PID controller tuning parameter (optimized factory settings) Proportional gain (output gain) Integral gain (error history) 2.Heater On/Off controlHeater On/Off control 3.Closed loop control:Closed loop control: On = automatic; Off = manual (requires output value) 4.Magnet set-point temperature ( ° C) - Must be set 10 ° C above ambientMagnet set-point temperature ( ° C) - Must be set 10 ° C above ambient 5.Submit: write settings to controllerSubmit: write settings to controller *Any change in parameters requires clicking the Submit button *Temperature plot history will reset

80. Temperature Page    Temperature sensor and heater output plots Sensor reading updated at 30s intervals Sensor reading updated at 30s intervals 1.Magnet set-point temperatureMagnet set-point temperature 2.Magnet temperature plotMagnet temperature plot 3.Ambient temperature plotAmbient temperature plot Includes heater/electronics dissipated heat Includes heater/electronics dissipated heat 4.Heater output plotHeater output plot 0-65355 full scale

81. The autoShim script The picoSpin 45 ships with factory set autoShim values. The Tx frequency and Pulse width values are specific to each instrument, all others fields contain general values typical for this script. autoShim control settings Plot window settings Plot window settings control the amount of data being plotted in the FID (top) and spectrum (bottom) preview plots in the web browser. Adjusting these values affect only the quality of appearance of the plots and not the acquisition or storage of data itself. Bringing to much data to the web browser requires more processing overhead which can affect the pulse sequence timing. 1. RF coil transmit (pulse) frequency in MHz: a magnet specific value 5. Quadratic shim coils shimming: Quadratic shim coils zx, zy, xy, z 2 and x 2 -y 2 4. Linear gradient shim coils shimming: Linear shim coils x, y and z 2. Max number of iterations of the simplex optimization algorithm 12. Main channel RF coil accessible via inlet port sample injection 7. Excitation pulse width corresponding to a 90° pulse: a capillary cartridge specific value 8. Number of points of the FID times series to acquire (maximum = 4096 pts) 9. FID zero filling of the “cost function” 13. Full bandwidth = 1/sampling period 14. Post-filter attenuation: at 4 kHz set to 10; at 16 kHz set to 8; at 64 kHz set to 6 10. Recovery time after switching coil from transmit to receive mode 11. Experimental 1 H relaxation recovery delay prior to executing a subsequent Tx pulse Enter an experimental “Run Name” to save script field variables and acquired shim/spectral data 3. Test run turns off optimization, allowing one to execute a single FID 0 iteration run 6. Cutoff criterion for terminating a shim based on current shim progress 15. Magnitude makes the signal always positive

82. Replace the Frit-filter on inlet fitting Refer to pS-45 Cartridge Maintenance Kit For detailed instructions on replacing the frit-filter

83. Replace the Frit-filter on inlet fitting