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MAN0410-1.1 Morphologi User Manual.book

1. e ill 7408 These are FireWire connection and cable connects the camera to the FireWire port on the computer The cable carries both power and data Trigger pulse connection and cable this is reserved for future use O Power LED this shows whether power is supplied to the camera The FireWire ports on the card in the PC are shaped like this ill 7680 0 PSU The small Power Supply Unit PSU supplies power to the instrument controls not the optical unit itself Caution A Do not use any other PSU with the instrument Morphologi G3 Page 3 13 Chapter 3 Page 3 14 Hardware features Joystick The joystick allows the user to move the microscope stage manually ill 7409 The components are XY joystick Two sliding tensioners Two hot keys to adjust the speed of the stage and the focus motor Digipot fine focus control These are described below XY joystick Use this as follows B Deflect the joystick left or right to move the stage in the X axis B Deflect the joystick back or forwards to move the stage in the Y axis B Deflect the joystick diagonally to move the stage in the two axes together The further the joystick is moved from the central position the faster the stage moves Note Q The logic of the software joystick control can be reversed using the Microscope Manager s Joystick menu This allows leftward movement of the joy
2. CE Diam Hs circ m CE Diam Particles M Circ Parti mor p Circularity and Particles Sample Name Salt minus oblates 3 SOP Name S 2 5x User Name Dfenton Date 09 November 2006 10 06 04 Particles Counted 990 Circularity Number Distribution All Particles Sorted by Circularity Descending o o o o o Circularity 0 996 0 996 0 996 0 996 03 04 05 06 OF Circularity 0 995 0 995 0 995 0 995 Circularity Mean 0 963 Circularity D n 0 1 0 927 Circularity STDV 0 029 Circularity D n 0 5 0 970 200 um Seale 1 100 Circularity RSD 2 96 Circularity D n 0 9 0 384 Ready Dfenton Morphologi G3 Page 8 19 Chapter 8 Page 8 20 Viewing the results The reports displayed depend on the workspace used There is a table in Chapter 12 showing which reports are displayed in each of the supplied workspaces Report parameters The reports for a single parameter all show the same statistics minimum maxi mum Relative Standard Deviation RSD and mean plus D n 0 1 D n 0 5 and D n 0 9 The Standard Deviation is shown where relevant The provided reports should be sufficient for most users but the user can create their own custom reports using the Report Designer as described in Chapter 13 Identifying data points On graphs which show many records it may be difficult to see which record each line represents Move the cursor over a line to show a display naming the record
3. 2d Malvern Sample Name Salt Glass Beads User Name jmalone Particles Counted 1080 Particle Size Convexity smoothed over 11 points SOP Name 8 2 5x Date 09 November 2006 10 06 04 Particle Shape HS Circularity smoothed over 11 points Morphologi G3 04 05 06 OF 08 098 i 1 04 05 06 Convexity HS Circularity Convexity Mean 0 986 Convexity STDV 0 010 Convexity RSD 1 03 Convexity D n 0 1 0 974 Convexity D n 0 5 0 386 Convexity D n 0 9 0 996 HS Circularity Mean 0 927 HS Circularity STDV 0 052 HS Circularity RSD 5 523 HS Circularity D n 0 1 0 859 HS Circularity D n 0 5 0 940 HS Circularity D n 0 9 0 370 scoombe Each report tab in the window names its associated report The tabs shown below would indicate that three reports Classification chart Classification table and Particles are available in the current workspace 4 B Records H Scattergram 27 Comparison f Classification chart IM cc Classification table IM ic Particles M User defined reports have their own tabs The M after a report name indicates a report supplied by Malvern Instruments To view a measurement as a report first select the measurement or group of meas urements in the Records tab then select the appropriate report tab Page 5 7 Chapter 5 Software Features Software components The main components of the Morphologi G3 window are labelled below an
4. Control module fuse this is user replaceable See the Essentials manual Connection panel The connection panel at the back of the instrument is shown below e ill 8155 The required connections are labelled as follows RS232 computer connection Joystick connection O Control module 40V DC power For information the other connectors are listed below Z motor connection STAGE XY stage connection Internal control connection Page 3 7 Chapter 3 Hardware Features RS232 stage controller connection Optical unit auxiliary interface connection D User changeable T5 0A fuse two spare fuses are supplied OD External PSU connection Caution A Do not plug unplug connectors while the box is powered on Page 3 8 MAM 0410 Hardware features Chapter 3 Morphologi G3 Integral SDU G3S only The SDU uses a laboratory s compressed air supply 5 5 bar minimum to 9 bar maximum or a dispersant gas bottle to disperse the sample onto the glass plate A bottle of dispersant gas can be supplied by Malvern One small gas bottle performs approx 1000 dispersions The dispersion is made using the software as part of an SOP or by using the command Measure Disperse Sample It cannot be fired manually Several plates can be pre dispersed in one session then the SDU equipment cleared away before making any measurements Chapter 4 shows how to prepare a sample for disper sion The components a
5. Note Left click and hold the mouse button down on the large image to view the original non thresholded image This helps decide the threshold to use When satisfied with the threshold click the 3 button Page 6 18 MAM 0410 Measurement tutorial Chapter 6 13 The Scan areas dialogue is displayed Use this to specify which part of the slide to scan SOP Editor New SOP File Edit Help peack 3 i A Sample details Pre measurement settings Measurement control Select areas to analyse Sample carrier Sample Dispersion Unit Illumination Optics selection 10x 3 5um 210um y Threshold Analysis settings Filters x Classification Post measurement settings Reports Drag rectangles to define new areas or drag the edge of an existing area to change the size and or position Selected analysis area Circular vi X pos mm Y pas mm 45 000 50 000 Radius mm 43 000 o Refine position before measurement Area mm 5808 805 Est number of particles 371799 Est time this optic mins 344 Est total time mins 352 Cursor position 7 2 The outer rectangle represents the whole slide The shaded green area within this is the current scan area The dot red in its centre is the origin In the Selected analysis area panel at the right of the window select Circu lar if using the SDU to disperse the sample or if a circular analysis area is required with any other plate
6. Classification Impurity 1 os Solon eae Ms a CE E SE ten a E word hy Oe NR Djs tee a ene uS ER VE zAcceptable E ee a Pe ANT INS ene qp 300 Number of Particles BE Record 31 Sample x2 B Scattergrams plots any two parameters against each other and provides a simple visual distribution overview of particle data Here 1s an example Scattergram Record 3 1 000 100 10 Length um 0 14 T 0 000 0 001 0 01 0 1 1 10 100 1 000 10 000 100 000 1 000 000 Area um For all graph types right click and select the Properties tab to set up the display This allows changes to the parameters shown axes the key fonts etc Lower and upper limits can also be shown on trend graphs The online help has full details Morphologi G3 Page 13 13 Chapter 13 Page 13 14 Creating custom reports Graph Properties dialogue For a Distribution graph the General tab allows a choice of graph type and how it is to be displayed i e as a histogram or curve Smoothing a running average can be applied as described in Chapter 8 For a Trend graph the Properties dialogue has a Properties tab for selecting the parameters to view Properties Properties Options Axis settings Font Upper Limits Lower Limits Trend Plats click an a cell to modify Parameter Axis Multiplier Record Y Axis Add Remove Parameter Record Ur p Against horizontal axi
7. Security guidelines Once security is enabled each user must log in when the software starts Once the user is logged in only functions matching their relevant permissions will be acces sible all other functions will be greyed out To change from one user to another without closing down the software the first user must select Security Logout and then the second use Security Login and enter the appropriate username password Note Q It is preferable to assign at least two users to the Administrators group The user s username and password should be stored in a secure location This is to safeguard against accidental lockout or deletion of permissions that may prevent the security settings being available Remember that after any changes to the security system adding users changing permissions etc the new changes must be saved by selecting User Save A dia logue will appear on exit reminding the user to save changes Setting up User groups All users must be a member of at least one user group Note Only users assigned to the Administrators group can add to or edit the user group properties Toadda new group 1 In the Security configuration dialogue select User New Group or dou ble click on an empty Groups row to display the Group Properties dialogue shown below Pege 10 4 MAM 0410 Security Chapter 10 Group Properties Gane OO O i i a Members Permissions C Configure communications set
8. Part 3 Appendices Specification Specifications Morphologi G3 Dimensions Morphologi G3 with stage Weight 60kg Height 700mm Width 440mm Depth 760mm Suggested desk 850mm x 2500mm space includes computer and two screens Size and shape measurement Size range Standard instrument 0 5um 1000um With 1X option up to 3000um Shape measurement Multiple shape parameters calculated for each particle and distribution generated on each parameter Parameters Circle Equivalent diameter Mean diameter Length Width Perimeter Area Aspect ratio Circularity HS Circularity Convexity Solidity Elongation Major Axis Max Distance SE Volume Mean Intensity Intensity Standard Deviation Page A 1 Appendix A Page A e Other parameters Specification Number of particles Limited only by available disk space but typically 5 000 analysed to 500 000 Sample quantity Typically 5mg to 200mg Validation and trace 21 CFR Part 11 compliant full IQ OQ calibration grat ability ings traceable to NPL Optical configurations The optical system is the Nikon CFI 60 Brightfield Darkfield system The specifi cation for this is as follows Magnification 1X at camera option 2 5X 5X 10X 20X 50X Approx total magnifi 48x 120x 240x 480x 960x 2400x cation on 17 screen Min particle size um 50 13 6 5 3 5 1 75 0 5 Max particle size um 3000 1000 420 210 100 40 Num
9. Viewing the new report gt To view a newly saved report in the main Morphologi G3 window 1 Select View Workspace Edit Workspace and select the workspace to associ ate the new report with In the Report Pages tab select the check box for the newly created report Click OK When the appropriate workspace is selected the report will be shown as a tab in the measurement file window Click the tab to see the report Note If a report that is currently displayed by the main application is edited sav ing the edited report immediately refreshes the view in the main window Any changes made are shown immediately Other information shown on a report The printed copy of a report shows the software version number and instrument serial number at the bottom of the page This is important information if a call is made to the Malvern Instruments help desk The name of the measurement file the report is based on is also shown Protecting a report A report can be locked with a password to prevent unauthorised changes gt To password protect a report 1 Morphologi G3 Open the report in the Report Designer and select Tools Protection Pro tect report A dialogue appears asking for a password to be entered and confirmed Write the password down in a safe place as it will not be possible to edit the report without supplying it Click OK Page 13 17 Chepter 13 Creating custom reports Page 13 18 MAM 0410
10. 2 Tu N v b x For greyscale images Intensity Mean ranges from 0 black to 255 white A plain mid grey object has an Intensity Mean of 128 Intensity mean 150 Intensity mean 95 Page e le MAM 0410 What is the Morphologi G3 Chapter 2 B Intensity SD the standard deviation of the pixel greyscale levels in the object i e Intensity SD Where I is the intensity value of pixel i N is the total number of pixels in the particle Intensity SD is reported in greyscale levels from 0 to 255 A uniform grey object like this would have an Intensity SD value of 0 Statistics presented Morphologi G3 The main statistics presented by the software in the Malvern reports are described here The statistics of the distribution are calculated from the results using the derived diameters D m n an internationally agreed method of defining the mean and other moments of particle size See British standard BS2955 1993 for more details m y Vd 73 m n 7 D n 0 5 D n 0 1 and D n 0 9 are standard percentile readings from the analysis D m n B D n 0 5 is the size in microns at which 50 of the sample is smaller and 50 is larger This value is also known as the Mass Median Diameter MMD or the median of the volume distribution The v in the expression D v 0 5 shows that this refers to the volume distribution This can be replaced by s for surface 1 for length or n f
11. 330 o a 7 M Aw 390 4 0 990 re J o o leo 0990 7 3890 0383 Jj 0 989 0 989 108 JA 05389 J 089 0 389 0389 0 388 0389 0 389 J e o ns989 o Oo osaga n38 oaee ee n3988 o o 0 o o o 4238 UM L 3238 w LL 328 es 388 JJ AE D 398 n388 _ Jede 388 a Oo o Oo E o 0 588 y 388 o g8 0a 388 A A ODE o DI X 388 0 38588 n 388 o o0 o v ill 7707 Agglomerates and other irregularly shaped particles can be identified visually from the images and also quantifiably by using a shape parameter such as circularity All agglomerates will have a lower circularity value than primary spherical particles so sorting and filtering on circularity will quickly identify the level of agglomeration in a sample Page e 16 MAM 0410 What is the Morphologi G3 Chapter Z This example shows needle shaped particles in another same sample 07 2 Ji 0712 0772 J 077 J 0712 J oie Or Jj ne jJ 72 de o JJ w OF Fle gt NM yl2 s OF X Q l2 de y pls 4 O12 FA Ole J x O12 Ak D He A FA ae fie o o AN i riz FR O12 FR Ole F wo Urli Drle sao Dda sao Drle o Il PR EA Jv Urla sao Ula Ul yo grl s ri ath Pd ep SORT DFi Drit uie DFi Orla uris FK JOM DE Fl2 A Ele Fiz s Diz A Urls F
12. Save As to save a copy of this SOP using a name that clearly identifies it as a development SOP e g add dev to the filename In the Measurement Control section check the Save measurement frames checkbox Save measurement Frames L IDacuments and Settings All Users DocumentsiM Ey Either accept the default location for the stored frames C Documents and Set tings All Users Documents Malvern Instruments Morphologi Recorded or choose another location In either case make a note of where the frames are being stored Select a suitable objective lens in the Optics selection of the SOP and make a note of it In the Threshold section choose a nominal threshold setting If unsure of the value to choose use the Estimate button to let the software choose a setting Page E 1 Appendix E Page E e Optirnising threshold settings 5 To save time use a smaller area than will be in the final SOP To do this in the Scan areas section adjust the area whilst monitoring the Est Total time field To speed up the process further set the Overlap to 0 in the Optics selection 6 Click OK to save the SOP then perform the measurement Measure SOP Note If previously saved frames already exist in the folder the system will prompt whether to overwrite them or not Analyse the frames using different thresholds Note Measurement from file is only available when connected to an instrument 1 When the me
13. www whatman com Mixed Cellulose Ester Membranes in the range ME 24 to ME 29 are suitable and available in both 25mm and 47mm sizes Page D 5 Appendix DO Optional sample plates Wet Cell The Wet Cell is a glass cell contained within a magnetically clamped metal frame that can be filled with a liquid sample It is designed for the measurement of sam ples whose particles are best dispersed in either suspension or solution The Wet Cell is suitable for measuring particles gt 15um and is only compatible with the 2 5X or 5X objectives due to depth of field restrictions at greater magnifications Part No Description MOR5200 Wet cell Overview The parts of the Wet Cell are shown below ill 8213 The features illustrated are Upper frame Lower frame Thumb recess Page D 6 MAM 0410 Optional sernple plates Appendix LJ 000009 Glass cell upper Glass cell lower Gasket Luer fittings Luer fitting caps Usage Assembly Before assembly always clean the Wet Cell thoroughly 1 2 3 6 Place the upper frame 9 face down on a suitable clean surface Insert the glass cell upper into the upper frame Lay the gasket on to the glass cell upper Place the glass cell lower on to the gasket sandwiching it between both glass sections flat side facing downwards with the Luer fittings upwards Position the lower frame B onto the upper frame encasing the entire glass cell betw
14. CE diameters for example Once a distribution has been generated all statistical parameters such as mean median mode standard deviation D n 0 1 and D n 0 9 percentiles etc can easily be calculated What is particle shape Describing a 3D particle size is complex but quantifying shape is even more com plicated There is an almost infinite number of ways to describe a complex shape and doing so seems to be deviating from the stated objective of reducing a sample to one quantifiable number So why measure shape Why is shape important Measuring size alone sometimes misses important but subtle differences between samples Some batches of samples may differ by such a small amount that this dit ference is lost during the translation to a CE or spherical equivalent diameter For example consider these three shapes Square of side Equilateral triangle of side Circle of diameter 2 units 3 039 units 2 257 units ill 7556 These shapes all have the same area four square units When they are converted to a CE diameter they give the same result 2 257 units Page e 3 Chapter 2 What is the Morphologi G3 This highlights the main disadvantage of measuring size only very differently shaped samples may be characterised as identical simply because they have similar projected 2D areas Particle shape often has a significant influence on final product performance parameters such as flowability abrasive efficiency bio a
15. Id parameter 2 7 Identification label 3 3 Illumination 6 12 Image capturing in colour 8 27 exporting 9 7 Image file 5 25 Image files dialogue 7 4 Image menu 5 24 Installation 1 5 instructions B 1 Instrument Lifting B 5 Instrument menu 5 24 Intensity Mean 2 12 Intensity SD parameter 2 13 J Joystick components 3 14 hot keys 3 15 reversing joystick logic 5 25 Joystick menu 5 25 K Kohler illumination 3 5 L Lamp house locations 3 4 two types described 3 6 Length parameter 2 9 Lifting the instrument B 4 B 5 Light Light calibration button 6 4 6 16 Light calibration command 5 24 Light intensity 5 21 5 24 6 33 6 35 Live picture window 5 19 Logging on 10 7 Logon 6 2 7 2 Lower Limits graph 8 23 8 24 Morphologi G3 Index Magnification choosing 6 5 specification A 2 Main window 5 2 Maintenance 1 5 Major Axis parameter 2 8 Malvern personnel 1 3 Manual exclusion tool 8 7 Manual lamp control 5 24 Manual Microscope See Microscope Manager Max Distance parameter 2 10 Maximal Feret diameter 2 10 Measure menu 5 11 Measurement tutorial 6 1 7 1 measurement 8 2 Measurement control dialogue 6 9 Measurement file exporting 9 4 in display 5 2 opening 6 28 overview 2 18 Measurement Manager described 5 16 progress meter 5 17 toolbar 5 17 using 6 28 Measurement record 8 2 Measuring a sample 6 28 Measuring from file 7 4 Menu bar Edit menu 5 10 File menu 5 9 Help 5 25 Image 5 24 Instru
16. Position Control Light source Step Size m a EPI Oo Move Frame Mave Z Z position mm 0 000 Move Target H DA Advanced Ie IET Goto Zera Position Bright Field If required focus using the joystick or Z Position Control part of this dia logue This focusing is not saved and may need to be repeated later It may be necessary to click the EPI button to switch to the top light for this step as the cross hair will help set the position Fit to window check box this option selected by default allows the user to navigate the preview image smoothly in order to identify a region of interest on the sample Once the region of interest is centred in the preview de select Fit to window This provides a closer view of the sample making it easier to obtain a more precise level of focus Use either the joystick or the X Y Position Control part of this dialogue to redefine the scan area by moving its origin This is often needed if a cover slip is used with an oil dispersion sample as it is hard to always position the cover slip in the same place If the SOP specifies the Fixed focus option for the objective manual focusing is not possible skip to step 14 Page 6 31 Chapter 6 Page 6 3e Measurement tutorial 12 If the Fixed focus option is not selected in the SOP the Focus on sample control dialogue appears as shown below Use either the joystick or the Z Position Control part of this dialogue t
17. Sample dispersion as part of an SOP sequence described in Chapter 6 Use the Sample Dispersion Unit dialogue to specify injection pressure and time also settling time Sample dispersion independent of an SOP this is useful as a series of sample dispersions can be made in one session and the plates stored ready for use The equipment can then be cleared away and the dispersed samples measured without the need to stop after each to prepare the next sample This technique uses the command Measure Disperse sample The online help for the com mand gives full details This is also useful during method development to determine the sample quantity needed the dispersion pressure to use etc Cleaning up After making a dispersion remove the sample cartridge and dispose of the used foils then clean the chamber thoroughly before the next sample is prepared Make sure there is no dirt grit on the air tight seal on the base of the chamber Page 4 6 MAM 0410 Preparing the sample Chapter 4 Sample preparation without the SDU Morphologi G3 This section is for W Morphologi G3S users who do not want to use the integral SDU for example when using samples on microscope slides B Morphologi G3 and G2 users This section gives guidelines on other sample dispersion techniques There are var ious ways to prepare samples for measurement including dry dispersions wet dis persions which are measured wet and wet dispersions w
18. The Unclassified percentage 1s Unclassified A B C Although the unclassified particles are excluded the percentage figure for these is still shown It should be clear that this means the total of the percentages can exceed 100 E Incolumn 4 unclassified particles are included making the total number of particles 1100 All percentages are calculated relative to this For example the percentage in Class B 18 2 is 200 1100 B Incolumn 5 particles in Class B and unclassified particles are excluded This makes the total number of particles in the included classes 800 All percent ages are calculated relative to this Here the percentage for Class A 87 5 is 700 800 The percentage in Class C 12 5 is 100 800 The percentage for Class B is 200 800 25 Again this shows why the sum of the percentage figures can exceed 100 To summarise m The percentages are based on the number of particles which are included m If anything is excluded the total percentage will exceed 100 W Unclassified particles can be excluded by right clicking and deselecting the Show Excluded Classes check box on the General tab MAM 0410 Viewing the results Chapter 8 Capturing colour images Morphologi G3 This section shows how to capture colour images of particles These can be saved into a graphics package printed out presented alongside a report emailed to colleagues etc They may also be useful if using polarisers During th
19. This scans a circular area The X Pos and Y Pos figures show the position of the origin of the scan area relative to the origin of the slide itself The Width and Height values show or set the size of the scan area The values below this show the estimated number of particles and estimated time needed to analyse this area the total figure includes any other objectives selected and the plate tilt correction Use these guidelines to decide on the area to scan depending on the number of particles needed for statistical signif icance and the time available Morphologi G3 Page 6 19 Chapter 6 Measurement tutorial There are two ways to specify the area of the slide to be examined m Drag the cursor to draw the scan area Multiple scan areas can be set up in this way by simply clicking and dragging to create new rectangles B Type values into the four position fields to specify the origin to start at and the dimensions of an area adjacent to this If merging multiple objectives and classifying types of particle by not just the distribution the scan area size must be exactly the same for each objective so type the values in rather than dragging an area The areas do not need to be in the same place Note Q The Delete button cuts the selected scan area from the display Select the Refine area at run time check box if required This allows an operator to set the origin of the scan area when they run the SOP 14 Clic
20. To add a class first click the New button and type in a name for the class over the text lt Classname gt Next click on the Parameter list and select a parameter In the Operator list select gt gt lt or Between For any operator except Between type in one value to complete this part of the definition For Between type in two values to define a range The and symbols for values allow successive ranges which do not overlap to be set up To exclude a class from the analysis select its Exclude class check box This acts as a more complex means of excluding particles than a filter as multiple parameters can be used to define the class Finally at the bottom of the dialogue select the Exclude unclassified parti cles check box if the measurement is to include only the particles which are members of classes We recommend leaving this unchecked during method development if this results in a high percentage of unclassified particles it sug gests that the classes need tuning Once the refined class definitions are correct this check box can be selected Click the button Click OK to close the Image File Analysis setup The Measurement Man ager opens for the file This window is initially blank for details of its compo nents see Chapter 5 Click Start The file being scanned will appear in the window as shown below The status bar will show the progress of the measurement Page 7 11 Chapter 7 Measure
21. 10 Morphologi G3 Class mismatches 8 25 Classes creating 8 10 meaning of percentage figures 8 25 overview 2 23 Classification Chart 8 25 Classification chart report 8 18 Classification dialogue 6 24 7 10 Classification graph 13 12 13 13 Classification tab 8 10 Classification table 13 15 Classification table report 8 18 Classification window 5 19 Clipboard copying 9 3 Colour image capture 8 27 Colour mode command 5 24 Colour mode command 8 27 Comparison tab how to use 8 14 results 8 16 Components 3 3 Computer specification 3 15 Configure export templates 9 10 Convex hull 2 11 Convex hull perimeter 2 5 Convexity 2 5 Convexity parameter 2 11 Copy Distribution Graph command 5 10 Copying with clipboard 9 3 Coverslip over sample 6 10 Creating reports 6 36 Creating SOP 6 6 6 7 D Darker button 5 21 Data Export Templates 5 12 Dendrogram 8 16 Dendrogram pane 8 15 Details window 8 6 Diascopic 6 12 Pace i Index Digital camera 3 13 Direct copying 9 3 Directories used for files 5 25 Dispersion quality 6 5 Distribution graph 8 21 13 13 Distribution window 5 18 Documentation button 5 17 Drag and drop 9 3 E Edit Documentation command 5 10 Edit menu 5 10 Editing SOPs 6 27 Elongation 2 6 Elongation parameter 2 9 Ensemble methods 2 14 Episcopic 6 12 Episcopic diascopic command 5 24 Essentials Manual 1 1 1 5 Example Results vmes 5 2 6 2 7 2 Exclude unclassified particles 8 10 Export to Clip
22. 1282 4 Inthe Threshold section choose a different threshold Typically try threshold settings and 10 of the first measurement For convenience record the threshold in the Sample details section Sample details Threshold Sample name Analysis settings Filters Classification My Sample Threshold 120 Notes Comments 5 Click OK and run the analyse from file measurement by clicking on the green start button The system now processes each frame in sequence At this point further measurements at different thresholds can be made by clicking on the Settings button choosing the Threshold section and then selecting a dif ferent threshold Remember to update the Sample name if this was used to record the threshold value Examine the results and finalise the SOP m When all the measurements using different thresholds have been completed click Close to close the Measurement from File window and select the Records tab Now look at the results from the different threshold settings For example select all relevant records in the Records tab and click on the CE Diameter tab to view an overplot of the measurements If all the records overplot quite closely this could indicate that the sample is not too sensitive to the threshold setting range used Alternatively it may be the case that at some threshold settings more fines are detected than expected If so this could be an indication that the thresho
23. 2 Perimeter This is equivalent to squaring the numerator and denominator of the Circularity calculation to obtain a more sensitive measure when comparing particles of similar circularity It is sometimes termed compactness A perfect circle has an HS Circularity of 1 0 while a narrow rod has an HS Circularity close to 0 Convexity the perimeter of the convex hull of the object divided by its perimeter The convex hull can be seen as the border created by an imaginary rubber band wrapped around the object In the diagram below is the added convexity area of the particle surrounded by the convex hull ill 7694 PerimeterofA B Convexity PerimeterofA The Convexity values are in the range 0 least convex to 1 most convex The Convexity is a measure of how spiky a particle is Solidity the object area divided by the area enclosed by the convex hull see above Solidity A B SE Volume the Spherical Equivalent SE Volume This is the volume of a sphere with the same CE Diameter see above as the object Page 2 11 Chapter 2 What is the Morphologi G3 Ye The SE volume measured in um is calculated as ill 7699 f 3 xx CEDiameter Volume 6 B Intensity Mean the average of the pixel greyscale levels in the object i e i N 2 Intensity Mean 1 4 Where I is the intensity value of pixel i N is the total number of pixels in the particle
24. Button Function Starts a new SOP Opens an existing SOP Saves the current SOP MS T Saves a modified SOP with a new name The following buttons are only available in the Filters and Classification dialogues Copy copies a selected filter or class to the Windows clipboard Paste copies a selected filter or class from the Windows clipboard K Undo undoes the last action gt Delete deletes the selected filter or class Some SOP Editor dialogues have tooltip help on their parameters To view this move the cursor over the YY symbol in the dialogue Morphologi G3 Page 7 3 Chapter 7 Measurement from files Measuring an image file This section explains how to measure a sample which is already saved in an image file To make a measurement from a file 1 Select Measure Image file This opens the Measurement Manager over laid by the following series of Image File Analysis dialogues Image files Sample details Threshold Analysis settings Filters and Classification 2 Use the Image files dialogue shown below to specify the file s to measure Browse for a file using the 3 button then use the button to add it to the Image files to analyse list Image File Analysis Image files Sample details Threshold Look in Analysis settings C Sample files G Filters Image files to analyse obitum Mane Modiied salte SJ Sample 1 Bmp 17 10 2007 14 793KB Bitr CNRC an
25. Chapter 13 Morphologi G3 Use the following buttons on the palette to add the listed elements to a report Button Use this to add Text Picture Frame used to split the report into sections De OFS Parameter Calculation TEN Particle image grid Graph use the list button to select a Trend graph a Distribution iol graph or a classification graph be Scattergram Classification table gt To add an element 1 2 Click one of the above buttons Move the cursor away from the palette When the cursor changes to a shape hold the left mouse button down and drag the cursor to draw a rectangle on the report This is called a marquee When the mouse button is released the marquee receives the selected element This will be marked initially by a hatched border like those shown below This shows that the element is currently selected e Report Designer VisionReportPage1 Screen Layout H File Edit View Layout Configure Tools Window Help DE EE Fes 3 19o 2 ee Osh SR x5 of oo oo obo pet oo oe fl oo fl oe os e Trend Graph Record Record Pe cc Fle bee eae ae Page 13 5 Chepter 13 Creating custom reports 4 Use the cursor to drag the selected element or black resize handles around its border to resize the element if necessary By default elements will automati cally snap to grid this means they will align t
26. Designer and placed in the default folder are automatically added to the list The default folder is C Documents and Settings All Users Shared Documents Mal vern Instruments Morphologi G3 Report Pages Editing a workspace To edit a workspace do one of the following B Click the button on the Workspace toolbar To edit a workspace other than the current one first select it from the list in the Workspace toolbar to make it the current workspace B Select View Workspace Edit Workspace and click on the workspace in the list displayed To change parameters use the Record View Parameters dialogue as described earlier Deleting a workspace If the current workspace is deleted the system uses the default workspace The default workspace cannot be deleted To delete a workspace 1 Select View Workspace Delete Workspace or click the workspaces list on the Workspace toolbar and select lt Delete Workspace gt 2 When the Select Workspace dialogue appears select the workspace to delete and click OK Select Workspace Select workspace Malvern Default v Malvern Default Sediments 4 Page 12 6 MAM 0410 Creating custom reports Introduction Morphologi G3 This chapter describes how to use the Report Designer to design custom reports These can display parameters which are not on the default reports Note The Malvern Instruments default reports described in Chapter 8 will be sufficient for most
27. G3S only LED displays Z limit switch Software controls The major hardware variables controlled by the software are Magnification Focus position Light intensity Choice of episcopic top light or diascopic bottom light illumination MAM 0410 Hardware features Chapter 3 Morphologi G3 Using the software to control these helps eliminate user and environment bias The aperture diaphragm lever Just below the microscope stage is a recess containing the aperture diaphragm lever This is used with the diascopic bottom light This should normally be set 80 90 open i e in the position shown below ill 7682 The aperture lock holds the lever in the 80 90 open position stopping it from being knocked out of position accidentally The lever is used to provide Kohler illumination an illumination technique that provides optimum resolution and contrast in the image Occasionally to get better contrast it s necessary to move the lever closer to the central 50 position If this is done always do the following B Add a note to the SOP pre measurement instructions m Add a note to the SOP post measurement instructions telling the operator to return the lever to the 80 90 open position afterwards LED displays Check the LEDs in case of problems LED Shows TX RX Communication with computer OK XYZ Run XY stage and focus electronics operational AUX Run Flashes when optical unit and
28. Measurement control Filters Sample carrier Sample Dispersion Unit Illumination Optics selection 5x 6 5um 420um Thrashed lt select gt Scan areas 10x 3 5um 210um Threshold Scan areas Analysis settings Classification Post measurement settings Reports Parameter Operator Value 1 Value 2 r i Use filtering to remove contaminants From the analysis Particles matching the filter criteria will be ignored The particles are still stored and can be viewed using the particle view Use this to set up filters for any of the reasons given in Chapter 2 Filters normally use OR logic meaning that a particle is excluded if it meets any of the filter criteria Filters using AND logic can be set up by defining a class see below then excluding it Note It may be better to make the first measurement without defining filters view the results then define filters using the Scattergram tab see Chap ter 8 and add these to the SOP Future measurements made with the SOP will then use these filters Filters Parameter Operator Value 1 Value 2 Circularity 0 600 Elongation Between 0 500 0 800 In the above example the first filter is defined to exclude all particles with cir cularity of 0 8 or over First select the parameter to filter on in the Parameter list In the Operator list select gt gt lt lt or Between Page 6 e3 Chapter 6 Page 6 24 Measurement tutorial For any operato
29. Oli Sx hil Aa Dril 5 ull Hbpo Z4 sow o Bri 5 Dril Kori JA d A oru JM on 7 0411 0711 v APE ti 0 711 0 711 Orii 71i i 0 711 0 711 OF s nli a Me a PA Cn FE OFfii aJ ozil 0711 y IP ee ee JA ill 7708 These can be detected visually using the actual images and also statistically by using a shape parameter such as Elongation Images of individual particles can be captured in colour this is described in Chap ter 8 Morphologi G3 Page e l17 Chapter 2 Wheat is the Morphologi G3 Morphologi G3 concepts This section describes some of the terms the Morphologi G3 uses The measurement file Each time a measurement is made the measurement data are saved to a measure ment file vmes This file contains the distribution data but not the individual particle data this is held for each particle along with the image itself in an img file Ifa measurement file is copied or emailed the images will not be visible unless the img files are also copied emailed A measurement file can also contain records produced by editing a record for exam ple by filtering out some particles from the original measured set This example shows part of a measurement file display Record Sample Name SOP Name Date Edited Particles CE Diameter Mean 1 Salt Glass Beads ES 09 November 2006 2 Salt Beads B 5 2 Bw O9 No
30. Opens the SOP Editor for Image File Analysis This lists available files for selection Disperse Sample lt SOP names gt Morphologi G3 Make a dispersion with the sample loaded in the SDU Presents a list of recent SOPs for selection Page 5 11 Chapter 5 Page 5 12 Tools menu Software features The Tools menu has the following options Command Report Designer Function Opens the Report Designer described in Chapter 13 Microscope Opens the Microscope Manager window Allows Malvern authorised service engineers to perform Engineering maintenance tasks The engineering screens are pass word protected Maintenance Settings gt gt Data Export Templates gt Line Styles gt ER ES Settings Used for characterisation see the Essentials Manual This has the following sub options Opens a dialogue defining the parameters and the format in which the records are exported Once a template is cre ated the measurement data can be exported to other soft ware packages such as Excel or Wordpad using File Export data Changes the colour and style of lines in report graphs Changes the Acrobat PDF output File Security and Audit Trail settings Options Security menu Sets the following but the defaults are normally best monitor setup one or two monitors and COM port grating information directories used positions of objectives in the nosepiece To prevent unauth
31. Particle stitching 2 21 6 14 Particles pane 8 5 8 6 Particles shown in green 8 6 Particles toolbar 8 7 Particles touching 6 5 Password changing 10 6 on report 13 17 pdf file 6 26 Percentages in classes 8 25 Perimeter 2 10 Permissions defined 10 2 list 10 5 Picture in report 13 7 Plate command 5 24 Polariser 3 15 Post measurement settings 6 23 6 25 7 9 Power switch 3 6 Powering on 6 2 Pre measurement dialogue 6 29 Pre measurement settings dialogue 6 8 Preparing sample 4 1 Printing batch of records 8 3 reports 8 20 Protecting a report 13 17 MAM 0410 Morphologi G3 R Record view described 8 2 Recording of images 2 16 Records deleting 8 2 moving copying 8 2 selecting 8 2 sorting 8 3 Records tab how to use 8 2 records and columns 5 3 Remote support 1 6 Report adding elements to 13 4 CE Diameter and particles 8 19 Convexity and HS Circularity 8 19 creating 6 36 13 1 13 4 file 8 20 locking 13 17 Malvern default reports 8 18 opening 13 4 page screen layout 8 20 printing 8 20 saving 13 16 setting up elements 13 7 standard 8 18 Report Designer introduction 13 1 opening 13 3 Report Pages folder 5 25 Report tabs 5 7 Report view report types 8 18 Reports dialogue 6 26 Result Data folder 5 25 Results exporting 9 1 9 4 viewing 8 1 Result under plot 8 17 Reversing joystick logic 5 25 Running an SOP 6 28 S Sample measuring 6 28 Sample carrier dialogue 6 10 Morphologi G3 Index
32. a region of interest The Source View is the original image and the Background separation is the image with the chosen threshold applied Clicking on the background and holding the mouse button down shows the particle in its original form In the Microscope Manager click the Light calibration button This ensures that the threshold chosen subsequently is correctly related to a stand ard intensity level Check the focus for the objective then select a frame which is representative of the sample MAM 0410 Measurement tutorial Morphologi G3 Chapter 6 Click Grab to return to the SOP Editor with the frame selected This returns to the SOP Editor with the selected image like this SOP Editor New SOP File Edit Help Qe OO Blt A Sample details Threshold l Pre measurement settings Measurement control Sample carrier Sample Dispersion Unit Illumination Optics selection 10x 3 5um 210um Threshold Scan areas Analysis settings Filters Classification Post measurement settings Reports Background separation Source view 15 ajal Get new image Threshold 162 Invert Use the lo button to select an area of interest Use the amp and Q buttons to zoom in and out if required In most cases clicking the Estimate button pro duces a satisfactory threshold If not satisfied with this result set the threshold manually as described below The Threshold slider looks like this Thr
33. dialogue is displayed Image File Analysis pack Ba lo X Image Files Sample details Threshold Classification Analysis settings Classification Parameter Operator Value 1 Value 2 lt Impurity gt Exclude class CE Diameter um gt 150 00 lt select gt select Acceptable Exclude class CE Diameter um v lt select gt Exclude unclassified particles Use this to set up any required classes as defined in Chapter 2 In the example above two classes named Impurity and Acceptable are defined using the CE Diameter parameter Note Q It may be better to make the first measurement without defining classes view the results then define classes using the Scattergram tab see Chapter 8 and add these to the SOP Future measurements made with the SOP will then have the classes set up Use this dialogue to set up any required classes as defined in Chapter 2 Class rules use AND logic meaning that a particle is only a member of the class if it meets all of the rules set up Particles can be a member of more than one class In the example below a class named Class A is defined using the Circularity and Elongation parameters Pege 7 10 MAM 0410 Measurement from files Chapter 7 Morphologi G3 Classification 10 Parameter Operator Value 1 Value 2 i Class A gt Exclude class Circularity 0 600 Elongation Between 0 500 0 850 lt select lt select
34. each particle makes to the distribution is the same a very small particle has exactly the same weighting as a very large particle MAM 0410 What is the Morphologi G3 Chapter Z The graph below shows the same sample measurement displayed on a number basis The same sample measured on a number basis shows a clear second peak of fine particles with a CE diameter of approx 60um For diagnostic or troubleshoot ing purposes the presence of fines could be very important in order to understand fully the manufacturing process so the extra sensitivity to fines of image analysis may be vital CE Diameter um smoothed over 11 points CE Diameter um Record 1 sample 2 Morphologi G3 Page 2 15 Chapter 2 What is the Morphologi G3 Recording of images The ability to visualise images of individual particles gives the user an extra level of verification that backs up quantitative data and helps optimise method development and sample preparation This capability is particularly useful for example for deciding whether an irregularly shaped particle is a genuine primary particle or an agglomerate of smaller particles Recorded images of all particles can then be sorted filtered and classified according to the user s requirements This example shows a selection of principally spherical particles Te Selected Particles MN Circularity OO m330 a nd 330
35. further configuration of the security system Pege 10 8 MAM 0410 SUP management Introduction A Standard Operating Procedure SOP measurement uses pre set parameters to ensure that measurements made on the same type of sample are made in a consist ent way SOPs are ideal for repeatedly measuring the same type of sample Using SOPs avoids the need to set the same parameters each time a measurement is made this is tedious and also risks introducing errors into the settings Chapter 6 showed how to create SOPs using the SOP Editor This chapter describes m Extracting an SOP determining the parameters used to create a record B Modifying an SOP modifying an SOP or creating a new SOP by editing an existing one and saving it with a different name WB Distributing an SOP making the SOP available to other sites Extracting an SOP Morphologi G3 The SOP used for a measurement can be viewed by selecting the record and click ing Edit Extract SOP or right clicking on the measurement record and select ing Extract SOP The SOP Editor opens with all the settings exactly as they were when the measurement was made The SOP can either be re used for subse quent measurements or modified and saved as a new SOP This is very useful for finding the fixed focus level used for a measurement Pace 11 1 Chapter 11 SOP management Modifying an SOP This covers modifying an SOP and also creating a new SOP by editing an
36. group to the Member of list 5 After adding the user to all required groups click OK To edit a user 1 To edit an existing user double click on their name in the Security Configu ration dialogue to display the User Properties dialogue 2 Proceed as described above for adding a user When a user logs on If security is configured the user is asked for a password by this dialogue when they try to log in 44 Type your user name ta lag an Malvern usw Morphologi G3 Page 10 7 Chapter 10 Security If the administrator selected the User Must Change Password at Next Logon check box see above the user sees this dialogue when they log in Set password Change your password User name Stuart Coombe Curent Password Confirm Hew Password EE To change their password the user has to enter the current password then specify a new password and confirm this Pressing OK makes the change to the security set tings If an administrator used the User cannot change password option this prevents a user changing a password once it has been initially set Otherwise once they have logged in a user can change their password using Security Change password To keep the system secure it is advisable for users to change their passwords regu larly to prevent unauthorised access The only possible exception is the security administrator s account where forgetting the password to this account could pre vent any
37. magnifications of 10X and below due to accuracy limits Page 6 14 MAM 0410 Measurement tutorial Chapter 6 The system displays the following warning if particle stitching is enabled SOP Editor 2 Particle stitching needs the Frame overlap For this optic to be set to 0 0 and works best with na particle segmentation ref Press yes to set the overlap and segmentation Far particle stitching Click Yes to continue Overlap is disabled and the overlap value is set to zero Specify Manual focus if it s necessary to let the operator set the focus at the start of a measurement Choose this if reusing an SOP designed for particles of one size range on particles of a different size the operator must adjust the focus for the new sample the first time they make a measurement Specify Fixed focus to force the system to focus a fixed distance from the sur face of the glass This increases the reproducibility of the measurement as it removes one area of possible operator bias It also speeds up the measurement process slightly It is necessary to focus manually the first time an analysis 1s made to do this use Edit Extract SOP and look at this dialogue to obtain the value Select Z stacking mode provides the option to choose whether the SOP should analyse Z stacked images rather than flat images To use this feature choose Enable Z stacking and then select the number of Additional layers above focus and optionally th
38. of the microscope Push the Polariser slider in gently with its nameplate and adjustment screw fac ing towards the microscope nosepiece There are two click in positions which can be felt as the slider is pushed in Push the slider into the second click in position To adjust the Polariser turn the adjustment screw To remove the Polariser from the optical path pull it out until it reaches the first click in position This aligns the empty hole with the optical path gt To use the Analyser 1 3 4 Remove the rubber cap from the ANALYZER slot on the right hand side of the microscope Push the Analyser slider in gently with its nameplate and adjustment screw fac ing upwards There are two click in positions which can be felt as the slider is pushed in Push the slider into the second click in position To adjust the Analyser turn the adjustment screw To remove the Analyser from the optical path pull it out until it reaches the first click in position This aligns the empty hole with the optical path Morphologi G3 Page 3 17 Chapter 3 Hardware features Page 3 18 MAM 0410 Preparing the sample Introduction Morphologi G3 Correct sample preparation is essential to obtain accurate measurements The objective is to disperse the sample and distribute it as a monolayer on a glass slide An image analysis system cannot produce good results from a poorly prepared sam ple so it is worth spending
39. paper gt To use Scan Area Composite 1 From the Scattergram tab view right click on any particle and click View area composite The Scan Area Composite window is shown Scan Area Composite Scan Area Composite Save Image This image is centred on the region in which the selected particle was scanned The blue shaded area indicates the section of the image displayed in the Scan Area Composite pane The Source view area of the window shows the wider scan area for the sample MAM 0410 Viewing the results Chapter 8 Any discontinuities shown in particles are due to stage accuracy and camera rotation as this view is simply a tiled presentation of all frames Particle stitching and Overlap obviate this issue in actual particle measurements see Chapter 2 for more information 2 To examine a different region of the scan area position the red outline over the desired area in the Source view area and single click to select 3 Usethe amp and Q buttons to zoom in and out of the image as required The image will automatically centre on the originally selected particle when zoom ing The shaded blue area alters in size accordingly 4 To save the entire composite image for further reference click Save Image The standard Windows Save As dialogue is displayed Morphologi G3 Page 8 13 Chapter 8 The Viewing the results Comparison tab Use the Comparison tab to compare results for a single morpho
40. report change too Frame Use the Frame tool to split the report into relevant sections With the element selected use its Properties dialogue to select the frame style etched raised sunken etc or change its colour Horizontal and vertical lines can be inserted into the report to divide sections To stop a frame overlaying other elements right click on it and select the Send to back option Morphologi G3 Page 13 7 Chapter 13 Page 13 8 Creating custom reports Parameter x Use the Parameter tool to display measurement variables such as CE Diameter SOP Name etc The parameter Properties dialogue has the same tabs as the text dialogue plus a General tab to specify what the parameter is as shown below Properties General Style Color Font Parameter Mo parameter has been selected 5elect a parameter eoa s 9 oa Measurement Audit H r Measurement Properties z Cancel I Arg Morphology 4 Particles S 4 Area pm zl T Area Pixels E 2 Aspect Ratio CE Diameter pm 4 CE Diameter Classes pum 4 IMZEBIETITS EB MED pm 4 CE Diameter Distribution so rt Statistics Se rl ree he f None Label only Both label EI Parameter arguments Column Title Argument Value CE Diameter D n pm On the Genera
41. the sample is polydisperse it is not possible to focus simultaneously on both small and large particles We recommend focusing on the smaller par ticles since a focus error on a large particle is less significant than focusing on a large particle and possibly losing the small particle altogether MAM 0410 Measurement tutorial Chapter 6 If necessary use the Goto Zero Position button to reset the Z position to its reference point using the Z switch When the particles are in focus click OK 13 The microscope goes to the gratings first then possibly the Z reference target before finally moving to the slide scan area 14 If tilt compensation self levelling is selected in the SOP the Live display window and monitor show the instrument focusing on the focus target like this ZS SOP Measurement general vsop v gH X Documentation Stop Help Close Particles 0 CE Diameter um Mean CE Diameter uim Std Dev 10 Percentile 50 Percentile 90 Percentile E Diameter um v volume Transformation Classifications long needles large oblates short needles large oblates Unclassified Number of Particles Multiview Result Video During this step which takes about three minutes the status bar says Level ling the plate slide The system checks each corner of the scan area in turn 15 If tilt compensation self levelling is not selected a single focusing cross appears for a sho
42. time optimising sample preparation conditions This chapter describes how to prepare samples for measurement It describes B G3S sample preparation using the integral Sample Dispersion Unit SDU B Sample preparation without the SDU useful for G3S users who do not want to use the SDU perhaps because a sample is already dispersed on micro scope slides and all G3 users The user can prepare samples manually with use of a separate sample prepara tion device or some other sample dispersion unit Note For the following methods several types of sample plate are available as described in Chapter 3 Select the correct plate for the sample and insert this into the stage Page 4 1 Chapter 4 Preparing the sample G3S sample preparation This section describes sample preparation on systems with the integral SDU The sample is placed between circular sheets of aluminium foil between two rings of plastic This assembly is termed the sample cartridge After sample is loaded in it this is placed in the top of a dispersion chamber There are two sample cartridge options WB Disposable the sample is loaded into the cartridge and dispersed then the cartridge is thrown away This avoids the need to clean the cartridge between dispersions B Reusable the cartridge foils are inserted by the user each time Each set can be used approximately 50 times This reduces the cost per measurement The dispersion chamber Th
43. to reject small dirt particles as noise but do not set it too high or parti cles may be lost forever In general it is best to leave this figure low and filter out unwanted particles later using filters Segmentation method Watershed segmentation is enabled by default This works well for spherical particles but not for example for crossed needle like particles Further segmentation methods suitable for different sample types will be developed in future Fill holes select the check box to include lighter areas within shapes When hole filling is turned on the middle of a doughnut shaped particle is counted as part of the shape When hole filling is on a doughnut shaped particle will have a high area without hole filling it will have a low area Size bands for speed the graph displays bins or bands rather than the raw data on each particle The default defines 1000 bands over the full range of the instrument This can be modified if more graph resolution is required in a specific range Click the Advanced button if it is necessary to change the size bands This dis plays the Size Bands dialogue see the online help for details Size bands Lower size um Upper size urn Fi P P P 2000 1000 Spacing Size bands um MAM 0410 Measurement from files Chapter 7 7 The Filters dialogue is displayed Image File Analysis Qeak Ba fox Image files Sample details Threshold
44. unlikely that one Page e 4 MAM 0410 What is the Morphologi G3 Chapter Z Morphologi G3 single shape descriptor will perfectly discriminate and characterise all applications and different combinations of shapes Notice that the long ellipse shape above dia gram top right has exactly the same circularity as the compact spiky shape bot tom left As a result a variety of shape parameters has been developed for the Morphologi G3 These provide a toolkit with different tools available as required for different applications For example an application concerned with perfectly spherical particles and meas uring perhaps for QC purposes the deviation from perfectly spherical would use circularity as the discriminating parameter However circularity would not be appropriate for an application with both spiky and elliptical particles present Two other commonly used shape parameters con vexity and elongation are described below Convexity Convexity is a measurement of the surface roughness of a particle It is calculated by dividing the convex hull perimeter by the actual particle perimeter The easi est way to visualise the convex hull perimeter is to imagine an elastic band placed around the particle The dashed line below represents the rubber band ill 7667 Convexity also has values in the range 0 1 Convexity 1 Convexity 1 Convexity 1 Convexity 0 7 Convexity 0 73 ill 7659 ill 7660 Page 2
45. uppermost 2 Detach the upper and lower frames by using the thumb recess O The mag netic clamp may require moderate force to break the seal 3 Remove both Luer caps and push out the glass cell Pege D 8 MAM 0410 Optional sample plates Appendix D 4 Separate both parts of the glass cell O from the gasket by sliding them apart preferably over some absorbent material Note CD The Wet Cell should be washed by hand only SOP configuration WB Select the Wet Cell Plate 100x80mm in the Sample carrier section of the SOP SOP Editor New SOP File Edit Help Qe 9Qi ngglpiusi Sample details Pre measurement settings Measurement control Sample carrier Carrier Sample Dispersion Unit Illumination Select the type of plate that will be used Optics selection Analysis settings Filters Classification Post measurement settings Reports Wet Cell Plate 100x80mm Options Compensate For plate tilt Coverslip over sample WB Inthe Optics selection Scan areas section of the SOP the system automati cally selects a rectangular scan area of the correct size for the entire cell This can be reduced if required Consumable parts Please contact Malvern Instruments to order the following replaceable parts Part No Description MOR4109 Consumable kit including Luer fitting caps and gaskets Morphologi G3 Page D 9 Appendix DO Optional sample plates Two Slide Plate The Two Slide Plate is use
46. users This chapter covers An overview of screen and page layout views what a report contains and how to start the Report Designer Opening an existing report Creating a new report Adding elements to the report Setting up the report elements Selecting elements Aligning and sizing elements Saving a report Viewing the new report Other information shown on a report Protecting a report Page 13 1 Chapter 13 Creating custom reports Overview This section introduces the Report Designer Views Two views must be created due to the different aspect ratios of the printed page and computer screen E Screen Layout shows the computer screen version E Page Layout shows the printed version This tailors the same contents to the dimensions of the printed page The two views do not have to be identical for example company logos can be added to the printed reports but not shown on the screen display Both views are saved in the same vrep file To change either layout right click on the screen and select Properties For the page layout this allows changes to paper size portrait landscape orientation and margins For the screen layout the screen size can be changed Report contents The report can contain one or more of any of the following items B Text user defined text B Picture graphics files can be added to customise the report B Frame used to separate parts of the report t
47. 5 Chapter 2 What is the Morphologi G3 A smooth shape has a convexity of 1 while a very spiky or irregular object has a convexity closer to 0 The shapes above show how convexity is unaffected by overall form a smooth needle has the same convexity as a smooth circle Elongation Elongation is defined as 1 aspect ratio or 1 width length As the name sug gests it is a measure of elongation and again has values in the range 0 1 A shape symmetrical in all axes such as a circle or square has an elongation value of 0 shapes with large aspect ratios have an elongation closer to 1 The shapes below show how elongation is unaffected by surface roughness a smooth ellipse has a similar elongation to a spiky ellipse of similar aspect ratio Elongation O Elongation 0 82 Elongation 0 ett Elongation 0 79 Elongation 0 24 Elongation 0 83 ill 7659 ill 7660 Conclusion The Morphologi G3 calculates a range of shape factors for use with different appli cations Three commonly used ones have been introduced here but as new appli cations are developed expect additional shape factors to be added to the list Along with the sensitivity gained from the ability to measure shape image analysis provides two other important benefits B Number based resolution B Recording of images These provide the user with additional information which contributes to a deeper understanding of the product or manufacturing proces
48. Axis Mean 89 26 ETT Mnt 200 300 400 50 Percentile 90 47 1 Major Axis 90 Percentile 162 75 v Volume Transformation Classifications EL 94 Ww lt Impurity gt x 4 i 4 lt Acceptable gt Unclassified 0 200 400 600 800 1000 1200 Number of Particles Multiview Result G video ill 7653 The components of this window are the following D Toolbar this is used to control the Measurement Manager Distribution window shows graphed values for the selected morphological parameter Summary window shows summary statistics for the parameter selected Classification window if classes have been defined shows the number of particles in each class Page 5 16 MAM 0410 Software features Chapter 5 Live picture window shows the sample frame This changes as the slide or plate is scanned Status bar shows instructions and information on the current operation in the measurement sequence Progress meter shows how far the measurement has progressed in terms of the number of particles detected and the estimated time before completion When a measurement completes its record 1s automatically selected in the Records tab The Measurement Manager window remains open ready to make further measurements Mode tabs three modes are available Where relevant these components are described in more detail below D Toolbar
49. Dim i s Integer Calc rrrr If Measurement S P SopType lt gt stInstrument nalysis Then Exit Function For i 1 To Measurement S0P Instrumentsettings Opticsettings Count If LenitCalc gt 0 Then Cale Calc Calc Calc HMeasurement S5 0P InstrumentSettings pticSettings Item il optic Next End Function MAM 0410 Creating custom reports Chapter 13 Morphologi G3 Particle image grid A particle image grid shows images of specified particles like this All Particles L 3 20 um Seale 1 1000 na L The image grid has a title bar a representation of each particle with a parameter below each the particle Id in the above example and a scalebar and number If the message The first particle is too big to display at this scale appears in the erid either drag the grid to increase its size or right click on it and use the View tab to change the scale To set up the parameter display right click on the grid and select Properties to open this dialogue Properties General View Font Sort by Parameter Sot Order Ascending t Descending i Show Parameter Values Classification lt All Particles Cancel Apply Help Use the Sort by Parameter list to specify what to sort on then use Sort Order to set ascending or descending order Deselecting Show Parameter Values allows more particles to be displayed in a grid To display just one class or unclassifi
50. Example 2 vmes DAR E File Edit view Measure Tools Security Window Help TE um 6 Ba ie oa di Combined reports ES E d ho i ER i ie 7 r v 7 MT MWT o V ry a 4 E Records 4 Scattergram 7 Comparison l CE Diam Circ Classification m _ CE Diam HS Circ M ik CE Diam Particles m f Morphologi CE Diameter amp HS Circularity Number Distribution 44h MAIN RN Sample Name Salt minus oblates 3 User Name Dfenton SOP Name 5 2 5x Particles Counted 990 Date 09 November 2006 10 06 04 Particle Size Particle Shape CE Diameter um HS Circularity 10 100 10000 i i j 05 06 07 CE Diameter jim HS Circularity CE Diam Mean pm 59 91 CE Diameter D n 0 1 um 26 52 HS Circularity Mean 0 929 HS Circularity D n 0 1 0 860 CE Diameter STDV yum 59 17 CE Diameter D n 0 5 um 40 55 HS Circularity STDV 0 053 HS Circularity D n 0 5 0 943 CE Diam RSD 98 77 CE Diameter D n 0 9 um 144 61 HS Circularity RSD 5 706 HS Circularity D n 0 9 0 370 Ready Dfenton 2 Circ Particles report This shows the Circularity data and also images of the particles themselves 7 Morphologi Example 2 vmes DEAR E File Edit view Measure Tools Security Window Help D a amp 3 e d Combined reports M isp iom a EE E ee V n Ee E CE Diam Circ Classification m
51. Filters Analysis settings Filters Copy o Undo X Delete Classification Parameter Operator Value 1 Value 2 Area Pixels lt 200 lt select gt lt select gt i Use filtering to remove contaminants From the analysis Particles matching the filter criteria will be ignored The particles are still stored and can be viewed using the particle view Use this to set up filters for any of the reasons given in Chapter 2 In the above example a filter is defined to exclude all particles under 200 pixels in area Filters use OR logic meaning that a particle is excluded if it meets any of the filter criteria Note that filtered particles can be removed after the measure ment if required Also note that AND logic filtering is possible by defining a class and excluding it Filters Farameter Operator Value 1 Value 2 Circularity 0 600 Elongation Between 0 500 0 800 First select a parameter to filter on in the Parameter list In the Operator list select gt gt lt lt or Between For any operator except Between type in one value to complete the filter For Between type in two values to define a range The and symbols for values allow successive ranges which do not overlap to be set up The Copy and Paste buttons can be used to copy filters between two SOPs When all filters are complete click the button Morphologi G3 Page 7 9 Chapter 7 Measurement from files 8 The Classification
52. For Scattergram tab components use Edit Copy Scattergram Edit Copy Scattergram and histograms or Edit Copy Particle window For Comparison tab components use Edit Copy Dendrogram Edit Copy Trend plot etc For a Report tab use the Edit Copy command for the relevant graph type Morphologi G3 Page 9 3 Chapter 9 Exporting results Exporting results To export a complete measurement file or a selection of records first open the required file and select File Export Result Use the File Parameter and Set tings tabs described below to select the exporting options File tab The File tab looks like this Export Data File Parameters Settings Export Options Export to file Export all records O Export to Clipboard O Export selection only File Options Export file path rn Instruments Morphologi Export Datalexported txt Append to file O overwrite File Select the appropriate radio button to specify the destination B Export to file writes to the file specified under File Options B Export to Clipboard copies the data into the Windows clipboard from which it can then be pasted into another application The File options selec tion box will be greyed out if this option is chosen Select the appropriate radio button for what to export B Export all records to export the complete measurement file B Export selection only to export only the selected records If exporting to an existing fi
53. GDOGCHACCNNM NN E TNR RN Salt minus oblates 3 5 2 5x 03 November 2006 10 06 04 True 990 59 91 0 929 0 952 Mean 4 6 437 133 06 0 862 0 840 Std Dev 480 71 81 0 061 0 038 RSD 110 54 04 7 061 Minimum 59 91 0 803 Dfenton The parameters shown are those defined by the workspace The tab shown above uses the Malvern Default workspace see Chapter 12 By default the statistics bar is displayed To disable enable it or any individual parameters select View Statistics bar then Hide or the statistics to hide Selecting records Records can be chosen using standard Windows procedures Either click a single record or hold down the Shift or Ctrl key and click to select multiple records Moving copying and deleting records There are several ways to handle the records m To copy one or more records into another open measurement file tile the two file windows then select the record s in one window Move the cursor towards the edge of the selection until a small rectangle and sign appear Hold the mouse button down and drag the selected record s to the other file window then drop the selection by releasing the mouse button W To copy a record select it then select Edit Copy Use Edit Paste to paste it into another file m To paste a subset of records copied to the clipboard using Edit Copy into a new measurement file select File New Measurement and name the file then use Edit Paste to paste the copied recor
54. ISML 1 1 How to use this manual 1 0 00 ee 1 2 Access to the instrument 0 00 eee as 1 3 ASSUMECINIONNANON s sisa eine a Al ie ee ee 1 4 Where to det helps vue x ae Re Dc Ro ad aaa ado doa atte ap V n 1 4 What is the Morphologi G3 IFCCS COM sailed sei eed ci ed ec de n decd oA Gide Gi Eo C IND EC 2 1 What the Morphologi G3 does llle 2 1 What are particle size and shape eee nn 2 2 Morphological parameters used llle nnn 2 7 Statistics presented u s aca hoe EY YEN es ea GR UE eG ae 2 13 Number based resolution lee hn 2 14 Hecording Of Images 1v 39 aor duit ax C acabe rata d Qu o do so 2 16 Morphologi GS COllOSplS 2 55 4 934 5 9 aca o ACA RC aee ACE SC AE ACE ES 2 18 Hardware features EEOC CHORD ord 0d o b ede oae Ret aba ndn acd tbt ied e qo esce a Sek 3 1 System configurations 3 9 edna ctc op CN Dh dio a E ae Se d 3 1 Kev Components ax eke Shee ea a acu X oaa Sd de eRe 3 2 TREASURER erne dr eoe o PE E ELA e oed o SR eat ey 3 3 Computer and monitors 0 00 eee hh ns 3 15 Polariser and Analyser options leeren 3 15 Preparing the sample Morphologi G3 INMOGUCTION amp ne2e cat X gro E dea doe d Ea MR ewe a m Bae eee 4 1 G3S sample preparation clle rn 4 2 Pagei Table of Contents Morphologi G3 Sample preparation without the SDU es 4 7 Software features Introduction adiac So Dee dls ERE Sas RS VERTO A 5 1 The ma
55. Intensity Mean Intensity SD Length um Magnification Major Axis 7 Max Distance Lim Perimeter nn SE valume Lm Solidiky Width m Value 101 299 12 0 340 IESE 45009 5339 j56355 537 0 973 1 000 0 060 0 946 514 38 109 15 209 ESSE esl 156 72 EI S ay 3031 55 1 000 20 01 MAM 0410 Viewing the results Chapter 8 Using the Particles toolbar The Particles toolbar is shown below Te show All Classes Id dis EU Its components are Control Use Expands the Particles pane to fill the entire window Controls what particles are displayed using classes or the selection rectangle Individual classes can be 3 Classes included using the list those ticked are displayed Shaw All Classes Show Only Selected Particles Unclassified Close The button text changes reflecting the selection Specifies the morphological parameter to sort on The default is to sort by particle Id Intensity Mean z iz l Sorts the particles in ascending or descending order T Ye This button toggles This toggles between showing and hiding all l excluded particles see Creating Particle filters below When filtered particles are hidden the button is ey 3 Produces a new measurement record taking account of the particles currently excluded The original record remains available in its unaltered state u Opens a Particle in Image dialogue showing the particle as
56. Limits Lower Limite General Graph Display Smoothing Asis settings Display Display Show multiple measurements Show volume transformation General tab This is the tab displayed above Its components are B Graph type controls the parameter shown This can be any of the morpho logical parameters Area CE diameter Circularity Elongation etc B Display this can be histogram frequency curve oversize or undersize curve plus combinations of these B Show multiple measurements select this check box to show more than one measurement on the graph B Show volume transformation select this check box to show volume based measurements Graph Display tab This controls where on the graph to show the key whether to show tips and whether to fill curves Page 8 21 Chapter 8 Page 8 2e Viewing the results Fonts tab Use this tab to set the fonts used for text on axes Axis settings tab This controls whether to show graticule logarithmic or automatically scaled axes Distribution Graph Properties Upper Limits Lower Lirnits Graph Display Smoothing xis settings Graticule Logarithmic Auto scale Smoothing tab The data on a Distribution graph are smoothed This means plotting a running average also termed a moving average All points included in the calculation are weighted equally This removes any spikes in the data These should not occur when l
57. Microsoft Excel specify tabs as separators Page 9 5 Chapter 9 Page 9 6 Exporting results Output example The following example shows the start of a comma separated file which includes a header row Record Sample Name SOP Name Particles CE Diameter Mean um Circularity Mean Convexity Mean Elongation Mean Length Mean um Width Mean pm Area Mean um I CRM Test JSOP 959 514 1 12 0 923 049069 0 2 lapl T4 40 124 99 cools A2 CBM Test SOP 20 7 so ely OO 02 9Gly On 4 69407720100 925422 Exporting particles Information on each particle in a single record can be exported as a text file This option is not available if multiple records are highlighted Select File Export Par ticles and use one of the following tabs to select the export options B File as described in the previous section but without the choice of all records or selection only W Sort use this to specify which parameter to sort the data on and whether to sort in ascending or descending order Export Data ea File Sort Classification Settings Particle Classification Export all particles in result na classification data CO Export particles in classes from selection below zImpuriby 2 M zAcceptable gt Include filtered particles in export MAM 0410 Exporting results Chapter 9 B Classification shown above particles in one or more classes can be exported There are options allowing export of unclassif
58. Morphologi G3 User Tierauiel MAMNOAIO Issue 1 1 August 2008 O Malvern Instruments Ltd 2008 Malvern Instruments makes every effort to ensure that this document is correct However due to Malvern Instruments policy of continual product development we are unable to guarantee the accuracy of this or any other document after the date of publication We therefore disclaim all liability for any changes errors or omissions after the date of publication No reproduction or transmission of any part of this publication is allowed without the express written permission of Malvern Instruments Ltd Head office Malvern Instruments Ltd Enigma Business Park Grovewood Road Malvern Worcestershire WR14 1XZ United Kingdom Tel 44 0 1684 892456 Fax 44 0 1684 892789 Morphologi is a registered trademark in the UK and or other countries and is owned by Malvern Instruments Ltd Malvern and the green hills logo are registered trademarks in the UK and or other countries and is owned by Malvern Instruments Ltd Windows 2000 and XP are registered trademarks of the Microsoft Corporation Morphologi G2 and G3 are registered trademarks of Malvern Instruments Eclipse is a registered trademark of Nikon Corporation FireWire is a registered trademark of Apple Computer Inc Printed in England Table of Contents Part 1 Operator s Guide Introduction to this manual INMt OOUCHON st esac ess Cons Resa Dee ew ee hese T
59. NO410 Issue 1 1 D ballez l instrument Auspacken des Gerates Desempaquetado del aparato Desempacotamento do instrumento Morphologi G3 Page B 1 Appendix B Unmpacking instructions If there is any sign of damage contact the freight carrier immediately Si vous constatez des signes de dommages quelconques contactez imm diatement le transporteur Bei irgendwelchen Zeichen von Transportsch den den Spediteur sofort benachrichtigen Si se advierte alg n signo de deterioro contactar inmediatamente con el transportista Contacte imediatamente a empresa transportadora se houver qualquer sinal de embalagem danificada Cut through the plastic bonding straps D coupez les courroies d attache en plastique Die Kunststoffoander durchschneiden Cortar las tiras de sujeci n de pl stico del embalaje Corte as tiras pl sticas que prendem o conjunto 3 Carefully cut the plastic packing tape along the fold lines D coupez avec pr caution le ruban d emballage en plastique le long des pliures Die Kunststoffverpackungsbander vorsichtig entlang den Falzlinien durchschneiden Cortar con cuidado la cinta de embalaje de pl stico por la l neas de doblado Com cuidado corte a fita de embalagem pl stica ao longo das dobras Page B e 4 Open the wings of the carton and remove the top foam insert Ouvrez les rabats du carton et retirez les blocs de mousse du haut Die Klappen des Karto
60. OP 1 Select File New SOP or click the 28 button to open the SOP Editor 2 The Sample details dialogue shown below is displayed initially SOP Editor New SOP File Edit Help onaluw Pre measurement settings Measurement control Sample name Sample carrier Sample Dispersion Unit Illumination Optics selection 20x 1 8m 100um Threshold Scan areas Analysis settings Filters Classification Post measurement settings Additional options Reports Notes Comments Prompt operator to change the sample details before the measurement begins Prompt operator to confirm the sample details when the measurement is complete 3 Typein the sample name and any notes associated with the measurement These appear on screen in a Pre measurement dialogue when an operator runs the SOP Morphologi G3 Page 6 7 Chapter 6 Page 6 8 Measurement tutorial To prompt the operator to add documentation before or after making a meas urement select the relevant check box es under Additional options We recommend selecting the first check box at least This adds a Documentation tab to the Pre measurement dialogue asking for details such as batch number from the operator When the dialogue is complete click the but ton The Pre measurement settings dialogue is displayed SOP Editor New SOP File Edit Help eack 1 A Sample details Dre measurem Measurement control Show these instructions when the SOP s
61. RO td C 2 Canadian Regulatory Information es C 3 VCCI acceptance Japan only esee e C 4 Optional sample plates Inirere U eN el eR MESE TTECCTUTTISTTTTITSTTO ROT PET D 1 mila EROR RETE TTL UU ILL TL PERILAELNEIUSTUTOM D 2 Web ellos dorso deber redi eee ee ts rv eg a D 6 TWO Slide Plate ded dee ruo teat aca end aes et Ad wed EU SIS d D 10 Page iv MAM 0410 Morphologi G3 Table of Contents Optimising threshold settings IFittOCU CT OND seia vee Ge ce E dodo NE B Ao deut GE ed ARE e e EE Co E 1 Morphologi G3 Page v Table of Contents Morphologi G3 Page vi MAM 0410 Peart 1 Operators Guide Introduction to this manual Introduction Morphologi G3 This manual covers the operation of the Morphologi G3 particle characterisation system In simple terms this 1s an automated microscope and a software package for control measurement and analysis The instrument measures the size and shape of a sample of particles presenting the data according to the user s needs The aims of this manual are to Explain what the Morphologi G3 is Explain in simple terms how the instrument analyses particles Describe the Morphologi G3 hardware and software components Explain how to prepare samples Explain how to use the instrument to make a measurement This describes analysis of both samples and image files Explain how to use export and report the result data Show how to perform
62. SDU operational 5V 12V Optical unit and SDU power is OK XYZ on XYZ and control system power is OK PWR Main power source is connected and optical unit switched on Page 3 5 Chapter 3 Hardware Features Back panel The back of the optical unit shown below gives access to the lamp houses power connection power switch and other connections ill 8122 The parts shown are Air inlet G3S only connect the air supply to the SDU here Reflected light lamp house this contains a 100W halogen bulb This light source is mainly used for autofocus and filter applications O Gas bottle clamps for the optional SDU dispersant gas bottle D Transmission light lamp house this contains a 100W halogen bulb This light source is used for most common applications Note The two lamp houses can be plugged in at the same time The only reason to remove one is to change a bulb as described in the Essentials Manual Main power input switch main on off switch for optical unit The fuses are mounted inside the rear panel If a blown fuse is suspected ask Malvern Page 3 6 MAM 0410 Hardware features Chapter 3 Morphologi G3 Instruments for a service engineer visit Operation of this switch also activates power to the control module so long as its PSU is plugged in Connection panel this is described in the following section Control electronics power inlet connect the 40V PSU cable here
63. Sample details dialogue 6 7 7 5 Sample plate types and components 3 12 Sample plate holder 3 11 Save measurement frames 6 9 Scan Area Composite 8 12 Scan areas dialogue 6 19 Scanning motion 2 19 Scattergram 13 5 in reports 13 13 Scattergram tab how to use 8 4 overview 5 4 selecting particles 8 4 Screen layout 8 20 Screen Layout reports 13 2 SE Volume parameter 2 11 Security administrator 10 3 guidelines 10 4 principles 10 2 user 10 6 Security menu 5 12 Segmentation method 6 21 Selected analysis area 6 19 Selecting elements in reports 13 15 Selecting records 8 2 Self levelling in SOP design 6 33 Serial number 1 5 13 17 Serial number label 3 3 Set intensity dialogue 6 34 Settings tab 9 5 Shape 2 3 Shape factors 2 4 Site requirements 1 5 Size bands 6 22 Slide selection 5 24 Smoothing 8 22 Software components 5 8 starting 6 2 7 2 Software modules 5 1 Solidity parameter 2 11 SOP creating 6 6 6 7 defined 2 19 editing 6 27 Page v Index extracting 6 27 running 6 28 saving 6 26 SOP Editor 6 6 6 7 7 3 SOP editor for image files 7 3 SOP mode Microscope Manager 5 23 SOP toolbar 5 14 Sorting particles 8 7 Sorting records 8 3 Source View 6 16 7 6 Specification A 1 B 1 Stage components 3 10 Standard deviation 8 20 Standard Operating Procedure See SOP Status bar 5 15 5 17 Summary window 5 18 Supervisor tasks 1 3 Surface Weighted Mean 2 14 T Table Classification 13 15 Template f
64. The Measurement Manager toolbar controls the measurement operation Its buttons are Button Es Settings Use Image file analysis only Opens the SOP settings which can be changed as necessary A Documentation Documentation an operator making an SOP measurement can click this and add sample information dispersion details etc Start Stop Accessory reserved for future accessory settings Start Stop these buttons start and stop the measurement Stop does not act like a pause button if it is pressed during a measure ment the user is asked whether to save or discard the particles measured up to this point Help Help opens help information on how to use the Measurement Manager Close Morphologi G3 Closes the Measurement Manager and returns to the Records tab If this is pressed while a measurement is in progress a dia logue asks the user whether to save or discard the particles meas ured up to this point Page 5 17 Chapter 5 Page 5 18 Software features Distribution window The Distribution window shows a graph of values for the morphological param eter selected using its drop down list It is constantly updated during a measure ment 2 0 0 5 0 0 0 1 1 10 100 1000 10000 CE Diameter um E Diameter um v C volume Transformation Smooth Graph The parameter can be changed at any time using the drop down list Select the Volume transf
65. This sound is produced by the micro stepper control and does not affect performance or reliability MAM 0410 Hardware features Chapter 3 Sample plates and the sample plate holder The sample plate holder is permanently mounted on the stage It is shown below o 6 l1 Orem o a 0 ill 8156 The features are 9 Rectangular recess for inserting sample plates see below for different applications The sample plate to use is specified during SOP creation Z reference target for offset correction and coarse focus alignment Z axis O Light calibration position for setting light intensity to a known level Four calibration gratings allow accurate calibration of magnification and focus Four gratings of different pitches are required to cover the full range of the instrument The gratings are etched chrome on glass and the measured line spacing is traceable to the National Physical Laboratory NPL The gratings are used to check calibration before a measurement The certified line spacing is printed on the slide as well as the certificate Caution A The grating pattern is printed on the underside to reduce the risk of dam age or contamination but treat the sample plate holder and plates with care Clean the plates as described in the Essentials Manual There are three sizes of sample holder depending on the sample plate used Morphologi G3 Page 3 11 Chapter 3 Page 3 12 Hard
66. adio Interference Regulations of the Cana dian Department of Communications Note that Canadian Department of Communications DOC regulations provide that changes or modifications not expressly approved by Malvern Instruments Limited could void your authority to operate this equipment This Class A digital apparatus complies with Canadian ICES 003 Cet appareil num rique de la classe A est conforme la norme NMB 003 du Can ada Morphologi G3 Page C 3 Appendix C Regulatory statements VCCI acceptance Japan only The Voluntary Control Council for Interference VCCI mark on this product sig nifies compliance to Japanese EMC regulations as specified by VCCI COREL PHUBLESERRSAEMHBMS VCCI DEH C4 24722AAT ESRIXIN OEIE C d CORE t RER CHATS CBR WEARZE TIE KSET TORAKA Au xpjEeiBd4 5k5SkK amp naccbbUutd ill 6793 Translation This is a Class A product based on the standard of the Voluntary Control Council for Interference by Information Technology Equipment VCCI If this equipment is used in a domestic environment radio disturbance may occur in which case the user may be required to take corrective actions Page C 4 MAM 0410 Optional sample plates Introduction Morphologi G3 Malvern Instruments offers a number of optional plates for the Morphologi instru ment For more information please contact Malvern Instruments This chapter provides detailed information on each optional plate t
67. administrator The administrator s then con trol access to the instrument by defining user groups and permissions B A user group comprises one or more users who have the same permissions WB Permissions are the access rights that are allowed for each user group these range from allowing SOPs to be edited to disabling view selection An administrator WI Creates users normally they assign each user a password although this is not mandatory If passwords are not set up the user just has to type their username to log in otherwise they must provide the password too WI Creates user groups users who share the same rights m Adds users to one or more user groups After installation The first time the software is run the security system will be disabled and an administrator user and administrators group will be created by the system This is so at least one user will have permission to configure the security system 21 CFR part 11 The security system can be upgraded to 21 CFR part 11 compliance by installing a feature key Once this is installed 21 CFR part 11 security settings can be applied and Audit trails can be viewed If the feature key is installed the normally grey 21 CFR 11 icon on the status bar becomes coloured This manual does not detail the 21 CFR part 11 option but concentrates on the standard security software Page 10 e MAM 0410 Security Chapter 10 Setting up the Administrator The first
68. arge numbers of particles are measured but can occur for smaller numbers of particles as may be found during method development To remove spikes from the data select the Smoothing tab and enable smoothing The Smoothing Span number specifies how many points are to be averaged ill 7671 MAM 0410 Viewing the results Chapter 8 Morphologi G3 The default smoothing value is 11 In the above example the feint line is the original data and the heavier line is a smoothed curve based on a smoothing span of 3 The first point on line point 3 on the X axis is the average of points 2 to 4 Smoothing only affects the visual appearance of the graph The original result data are not changed by it and calculated parameters such as D n 0 5 are based on the original data Note L As a general guideline if result data do show spikes it may be necessary to review the method used to increase the numbers of particles detected The Geometric Standard Deviation GSD of the sample influences this strongly For example to measure a sample distribution with lt 5 error the number of particles required is approximately 8000 where the GSD is 1 3 and over 60 000 for a GSD of 1 6 Upper Limits Lower Limits tabs These are mainly for use with trend graphs see below Trend graphs The configuration of a trend graph is controlled by this Properties tab Trend Graph Properties Properties Options Axis setti
69. asurement has completed choose Measure Image File In the Image Files section browse to the folder noted previously where the saved frames are stored Note this folder location is saved between sessions 2 Select all or just those required captured frames and transfer them to the right hand panel If using just a selection of images it may be useful to examine the images in the Image details preview area to ensure that they contain a repre sentative sample of particles If the preview proves too small for detailed inspection double click an image from the left hand panel to enlarge it using the default Windows Picture and Fax Viewer 3 Using the table below for reference enter the pixel size based on the megapixel resolution of the instrument s camera Generally G3 instruments have a 5 megapixel camera and G2 instruments use a 2 megapixel camera If unsure of the megapixel count of the camera the Image details preview area lists the width and height dimensions of the image files multiplying these together gives the number of megapixels Magnification Pixel size 5M Pixel size 2M 1X 28 4 4 2 5X 1 12 1 760 5X 0 560 0 880 10X 0 280 0 440 20X 0 140 0 220 50X 0 056 0 088 MAN 0410 Optimising threshold settings Appendix E Morphologi G3 In the example below the pixel size has been entered for the 10X for a 2 meg apixel image Pixel size Microns per pixel Size W x H pixels 0 440 s 1616 x
70. asurement settings Reports 4 Slide Plate 75x25mrn Options Compensate for plate tilt Coverslip over sample Under Carrier select the slide plate to use in the list an image of the carrier 1s shown for verification If making quick test measurements using a 2 5X objec tive or using a 5X objective in just a small scan area consider deselecting the Compensate for plate tilt check box to speed up the process a little We rec ommend using tilt compensation for all other measurements Coverslip over sample should be used whenever a coverslip is used on the slide by selecting this option the system knows how to focus to the correct Z position If a cover slip is used and this is not checked a focus error may be displayed during the measurement Click the button Page 6 10 MAM 0410 Measurement tutorial T SOP File Editor New SOP Edit Help Qu OLN Alta H Sample details Pre measurement settings Measurement control Sample carrier Sample Dispersion Unit Illumination Optics selection 20x 1 8pm 100pm Threshold Scan areas Analysis settings Filters Classification Post measurement settings Reports The Sample Dispersion Unit dialogue is displayed Foil Type 6 um Foil Injection Pressure bar Injection Time rns a Settling Time sec J Injection Volume Indicator Chapter 6 If using the integrated SDU check the Use SDU box Morphologi G3 Foil typ
71. ations in product that are associated with operator plant proc ess line etc Page 5 5 Chapter 5 Software features ch lesi vmes Hecord 6 A5 Kun amp Pu ersten lari Lluttes bur Vaihi O Ana prs CO CE Career pm O Length um ares Burd CO Max Diraca uri CO Panmeter urn C SE Vikan anit Foes und Deth jm COuMupaet Plate R2 AS Run 2 CO Creu C Caiway C Elengstion qom monacus OHS Circuir 5 Solid Rigas Pun f C Hn ensity Hegn 5 Intensity SD B ta 02 3 oa OS Ob OF i 11 zs m 245 Record Dijerat Inbensiby SD b eradty i F1 A5 run d DEenbon ill 8158 The components described in detail in Chapter 8 are D Q Page 5 6 Records selected shows the records selected in the Records tab before the Comparison tab was selected Selection area use this to select the record groups to use Records can be grouped initially by the user then the groups changed on the basis of the results displayed Parameter Variability radio buttons used to specify the morphological parameter to display Use the radio buttons to specify which parameter to base the clustering on Parameter Variability bar charts show the morphological variability of the selected records The widths of the bars show which parameter differenti ates the most Dendrogram clusters the selected records showing the degree of similarity between them Trend plot th
72. ays Instead of setting up sets of largely identical parameters each time copy an existing SOP and just change the required parameters This reduces the risk of making errors in the settings Scanning motion and frame overlap To guarantee that all particles in a given scan area are detected and no bias is gener ated towards smaller particles a frame overlap procedure is used Particles touching the edge of a frame are rejected and detected in the next frame instead The XY coordinates of each particle are logged and used to ensure that the same particle detected in consecutive frames is not double counted The examples below illustrate the importance of this mechanism Example 1 No overlap A B C ill 7446 The large black rectangle at the top represents the total scan area and this is scanned by three adjacent frames A B and C The tiered rectangles below this show the particles each frame will measure assuming that the two particles which fall on the lines between these frames are excluded Page 2 19 Chapter 2 What is the Morphologi G3 This system obviously generates a bias towards smaller particles as large particles have a higher probability of touching an edge To avoid inaccuracies caused by edge effects a degree of overlap is used This is shown by the example below Example 2 With overlap Here there is an overlap between frames so five frames are now needed t
73. b Morphologi G3 Page 6 e9 Chapter 6 T Page 6 30 Measurement tutorial If a carrier plate that has more than a single slide filter position is in use see Appendix D the user 1s prompted to select the relevant position s which are then displayed in green in the dialogue Pre measurement Slide s Select the slide positions to measure There are 1 slides required by the SOP Sample name sample manual measurement Comments O Select the first slide to use by clicking on it Next type the Sample name for this slide in Repeat this for each slide being used The SOP s Measurement control settings specify how this produces the result If multiple slides are selected for example this can produce a record for each slide or one combined record Click OK If the integrated SDU is being used the system checks that a plate is in position and then disperses the sample Wait for the settling time to pass The Microscope Manager performs initial focusing using the plate holder s gratings and Z reference target see Chapter 3 for details The Live Display window and the second monitor show these briefly The text Finding the focus reference point is displayed during this step MAM 0410 Measurement tutorial Chapter 6 Morphologi G3 10 If Refine area at run time was selected in the SOP the Refine scan area 11 position dialogue appears Refine scan area position amp Y Position Control
74. basic challenge of particle size analysis how to describe a 3D object with just one number Circle Equivalent CE diameter Image analysis captures a two dimensional 2D image of a 3D particle and calcu lates various size and shape parameters from this 2D image One of the parameters the Morphologi G2 calculates is Circle Equivalent CE diameter the diameter of a circle with the same area as the 2D image of the particle Particle shape of course influences this CE diameter but it is at least a single number that gets larger or smaller as the particle does and is objective and repeatable The 3D image of the particle is captured as a 2D image and converted to a circle of equivalent area to the 2D image The diameter of this circle is then reported as the CE diameter D of that particle as shown below MAM 0410 What is the Morphologi G3 Chapter Z Morphologi G3 JH 3D particle Captured as a 2D Converted to a Diameter of image circle of same area circle measured ill 7440 Of course a single value from one particle is not appropriate as the magic single number for management purposes It is unlikely to be statistically significant as the single value depends upon which individual particle is chosen A number of parti cles which are representative of the sample as a whole have to be measured and sta tistical parameters generated A more appropriate single characterisation number would be the mean of all the
75. board 9 4 to file 9 4 Export Data dialogue 9 8 Export data folder 5 25 Export template creating 9 8 parameters 9 5 Export Templates folder 5 25 Exporting images 9 7 Exporting particles 9 6 Exporting results 9 1 9 4 Extract SOP command 5 10 Extracting SOP 6 27 F Feret diameter 2 10 File adt 5 25 del 5 25 8 2 edf 5 25 img 5 25 pdf 6 26 txt 5 25 vmes 2 18 5 25 vrep 5 25 13 2 Vsop 2 19 5 25 6 26 Pace ii Morphologi G3 exporting to 9 4 measuring from 7 4 File menu 5 9 File tab 9 4 Fill holes 6 21 Filter plate D 2 Filtering particles 8 8 Filters overview 2 23 FireWire port connection 3 13 Fit to window 5 23 Fixed columns 8 3 Focus control joystick 3 15 Focus on sample control dialogue 6 32 Focus position 3 4 Focusing the microscope 6 32 Folders used for files 5 25 Fonts graph 8 22 Four slide plate 3 12 5 24 Frame overlap defined 2 19 setting 6 14 Frames in reports 13 7 Frequency curves 8 17 Fuse stage controller box 3 8 G Go to particle button 5 21 Grab button 5 23 6 17 Graph Classification 13 13 distribution 8 21 in report 13 12 modifying 8 21 trend 8 23 Grating description 3 11 Greyscale levels 2 13 Group Properties dialogue 10 4 Group See User group H Hardware components 3 1 Health and safety 1 5 Help desk 1 5 Help menu 5 13 Hole filling 6 21 Home Nosepiece 5 22 MAM 0410 Morphologi G3 Hot keys 3 15 HS High Sensitivity Circularity 2 4 2 11
76. cal distributions and clusters the measurements according to the parameter that varies the most In the Parameter Variability pane bar charts indicating the amount of varia bility are shown The parameter with the greatest variability is shown in bold and its radio button is selected by default Note that the distributions of the morphological parameters are compared not statistical values such as the mean In the Dendrogram pane the results are clustered according to their similar ity based on the distribution parameter selected by the radio button In the dendrogram short horizontal bars indicate similar records The horizontal dif ferences scale varies from 0 distributions overlap exactly to 1 no overlap of the distributions at all The software colour codes the clusters into a best estimate pattern The best number of clusters found is shown in the Clusters drop down list In the Trend Plot pane the mean value of the selected parameter is plotted to correspond with the dendrogram clustering This indicates what a Statistical Process Control SPC chart would look like for this parameter The X axis can be used to decide what pass fail criterion to set Finally the frequency and undersize plots are shown to indicate what the actual distributions of the clustered records look like The distributions are col our coded to match the clustering colours The clustering automatically found may not match the expected clusterin
77. can show the number of particles in each class the percentage in each class and the percentage unclassified as shown below Classes are set up in the SOP Editor Classification dialogue or preferably by using the Scattergram tab Classification Glass Spheres Salt Crystals Glass Oblates Unclassified 400 500 Number of Particles B Record 7 Salt minus oblates 3 To select a percentage display right click and use the Properties dialogue s General tab Class mismatches If multiple records are selected but these do not have classes defined in the same way the message Some records are not displayed due to mismatched classification settings is shown in the chart title Percentage displays The percentages shown when classes are excluded require explanation The second column in the following table shows the number of particles in each of three classes A B and C and the number of unclassified particles in this sample Morphologi G3 Page 8 e5 Chapter 8 Page 8 26 Viewing the results Number With Unclassified Class B and of Unclassified not Unclassified Class particles excluded excluded excluded A 700 70 63 6 87 5 B 200 20 18 2 25 C 100 1096 9 1 12 5 Unclassified 100 10 9 1 12 5 Total 1100 110 100 137 5 The remaining columns show the percentages that would be displayed in the chart under different conditions Note the following B n column 3 the sum of A B and C is 100 70 20 10
78. ces for use by customers 24 hours a day seven days a week Resources include software downloads frequently asked questions a knowledge base and application notes plus information on other particle characterisation solu tions that Malvern Instruments can provide MAM 0410 What is the Morphologi G3 Introduction This chapter describes What the Morphologi G3 system does The fundamentals of particle size and shape Definitions of the morphological parameters used The importance of number based statistics The benefits of image recording Morphologi G3 concepts the measurement file Standard Operating Pro cedures SOPs scanning motion and frame overlap filters and classes What the Morphologi G3 does The Morphologi G3 instrument provides the ability to measure the morphological characteristics size and shape of particles The particles can be on the microscope stage or in existing image files The instrument is a high sensitivity high resolution analytical tool for differentiating and characterising particulate samples Morphologi G3 Morphologi G3 image analysis provides number based statistics This means data are generated on individual particles as well as on the sample as a whole This makes the technique particularly suitable for detecting the presence of fines or for eign particles In simple terms 1 2 A sample is prepared and placed on the measurement slide or plate The sample
79. ch is valid Page 5 e3 Chapter 5 Software features m A series of messages is displayed in the bottom left corner of the window These give some status information and instructions on how to proceed Menus The Microscope Manager menus are described here Image menu This has just one command Command Save as Instrument menu Function Saves the current image as a graphics file bmp jpeg etc for use in reports To include a scale bar in the image check the Add scale bar to image box in the Save dia logue This can also be used to save coloured images as described in Chapter 8 The commands are the following Command HMU Safe Function Not used on the Morphologi G3 Episcopic Switches between diascopic and episcopic illumination Manual lamp control Morphologi G2 only turns off software control of the lamp intensity enabling the front panel Lamp intensity dial to do this instead see Chapter 3 Plate Tells the software which plate is in use Slide Light calibration Colour mode White balance Page 5 24 Selects the slide to use 1 to 4 on a four slide plate Resets the light intensity This may be necessary after a change of objective Used to take colour pictures of images see Chapter 8 Used when taking pictures of images see Chapter 8 MAM 0410 Software features Chapter 5 Joystick menu This has two commands Command Function Rever
80. cuments Malvern Instruments Morphologi If all the measurement data including the individual image files are required save the contents of all sub folders of the following folder This will allow completely new analysis of the data in future if required C Documents and Settings All Users Shared Documents Malvern Instruments Morphologi Image files are stored in the following folder by default should the user need to save or copy them to other PCs separately C Documents and Settings All Users Documents Malvern Instruments Morphologi Recorded To restore the files simply copy them back to their respective locations Page 5 e7 Chapter 5 Software features Page 5 e8 MAM 0410 leeasuirernernt tutorial Introduction After reading this chapter a user should be able to make simple measurements It begins by describing how to get started powering on the system starting the soft ware and placing the sample It then covers the following five steps to making a measurement Morphologi G3 1 Method development checking the sample dispersion choosing a magnifi cation and ensuring sample dispersion quality Creating a Standard Operating Procedure SOP setting up and saving measurement parameters to ensure that measurements of similar samples are consistent This optimises all software and hardware variables This is useful in Quality Control environments All variables are captured in a single SOP file t
81. d described in the sections which follow D Tabs used as described above to switch between the Records tab Scatter gram tab Comparison tab and Report tab Menus give access to commands Toolbars provide shortcuts to commonly used commands Statistics bar Status bar and 21 CFR 11 icon LEJ Ele Edit View Measure Tools Sec ty Window Help Bx Emi Ld rd Ha KS d Ba la P i Classification Particles 4 I R See J UT NN lecords Scattergram 1 7 Comparison cs Classification chart M aede i c C table fei Particles M b l Record it Sample Name SOP Name Date Edited Particles Class ount Glass Spheres Glass Spheres Class 1 Salt Glass Beads 5 2 5x t November SUUN 04 3 Salt Glass Beads2 BEAN E ione 20 2006 10 06 emm Mean 3 5 Std Dev 53 RSD A 37 Minimum 113 Maximum Denon ill 8159 The menu bar contains the main menu headings for all software functions The menu commands are described below Note the following B Items which end with a row of dots open a dialogue box B Items which end with an arrow gt present a list of sub options when the user clicks on the arrow B Items shown in grey are not available at the time This indicates that a security setting has been activated or the item is not relevant in the current context This applies for example if a user starts the software with no microscope con nected or powered on the Tools Microscope
82. d by the new workspace The command View Workspace does the same as the toolbar select the workspace name from those listed Note D If the Workspace toolbar is not displayed use View Toolbars and click Workspace Toolbar to turn its display back on Creating workspaces It is good practice to create different workspaces for different types of analysis To create a new workspace 1 Select View Workspace Add Workspace This displays the prompt for the Workspace name Workspace Name 2 Type ina name for the new workspace and press OK This displays the Work space parameters dialogue shown below This has two tabs Record View Parameters and Report Pages 3 Use the Record View Parameters tab shown below to specify which param eters to show in the Records tabs The list on the left displays all the available parameters for a record The parameters are arranged in folders to help navigation For example all the shape data are held in the Morphologi folder Morphologi G3 Page 12 3 Chapter 12 Workspace management Record View Parameters Report Pages Ei r Measurement ct Details Image DB H g Measurement Audit H A Measurement Properties H A Morphology rt SOP Settings it e gt Fixed columns 4 Specify the number of Fixed columns those which cannot be used for reor dering the list of records To add a parameter select it in the tree on the left and click the button to copy
83. dialogue is displayed Image File Analysis Qs Image Files Sample details Background separation Source view 100 Analysis settings Filters Classification E Image file Sample 1 Bmp Threshold 0 64 128 192 255 E Tal The Source View is the original image and the Background separation is the image with the chosen threshold applied A rectangle in the Source view panel initially in its top left corner shows the area which appears at a larger scale in the Background separation area Drag this around to look at other areas of the slide Clicking the Full screen button displays this panel at full size on the second monitor First use the button to select an area of interest and then use the amp and el buttons to zoom in and out as required In most cases clicking the Estimate button produces a satisfactory threshold If not satisfied with this result set the threshold manually as described below The Threshold slider looks like this Threshold 0 4 126 192 259 0 Gi g 247 EE 8 Page 7 6 MAM 0410 Measurement from files Chapter 7 Drag the 4 button to slide the Threshold bar to change the applied threshold level Aim for a threshold level which correctly identifies the particle edges This is the point where a light grey halo starts to appear around each particle The two numbers and define the range selected To contract the range itself use the 4l la
84. dified NjiSample x2 6 1KB Bitmap Image 17 05 2006 11 59 A Sample X2 1 1KB Bitmap Image 17 05 2006 11 59 N Sample X2 2 1KB Bitmap Image 17 05 2006 11 59 N Sample X2 3 1KB Bitmap Image 17 05 2006 11 59 SI Samnle Y 4 1KR Ritman Imane 17105i2nn amp 11 59 Page 9 7 Chapter 9 Exporting results This does not produce coloured images for instructions on doing this refer to Chapter 8 Creating export templates For use with the File Export result command an export template can be created This controls the format record and particle data is exported in The template is stored as an edf file in the folder Export Templates gt To create a new export template 1 Select Tools Settings Data Export Templates to display this dialogue The buttons in this dialogue are Button Function Creates a new template Deletes the selected template Edits the selected template To change the name of a template double click on it and type a new name 2 Select the EJ button and type in a name for the template 3 Click the Es button This displays the Export Data dialogue shown below Page 93 8 MAM 0410 Exporting results Chapter 9 Morphologi G3 Export Data Settings Parameters Format Options Include Header row Enclose text in quotation marks Use tabs as separators O Use commas as Export Data Settings Parameters Measurem
85. ds into it B To remove selected records from the measurement file use Edit Delete The records will not be deleted from the software but stored in a file with the same name as the original measurement file but with the extension del This file is Page 8 2 MAM 0410 Viewing the results Chapter 8 placed in the Result Data directory If required the deleted records can be copied back to the original file Records can be selected either by clicking the cursor on the required record or by using the Navigation toolbar press its arrows to move up and down the list Sorting records To sort measurement file records in the window into alphanumeric order click on a column header to list all records in order based on the value of that parameter Click on the column header again to toggle between ascending A and descending V order A symbol in the header shows which of these the order is like this Mean x Sorting cannot be performed based on fixed columns those which are shaded grey How the workspace controls the Records tab The parameters shown the columns are specified when a workspace is set up using the View Workspace command see below The workspace used specifies which parameters appear in the records their order and whether any columns are fixed Workspaces can be selected created or deleted using the View Workspace commands or the workspace toolbar shown below Malvern Default Malvern Default i
86. e the default selection is 64m but you can also select 25m Injection Pressure bar the default is 0 8 bar for 6um or 4 bar for 25um but increase this for more aggressive dispersions Injection Time ms the default is 20ms for 64m or 10ms for 25um Settling Time sec this depends on particle size but defaults to 60 sec onds Increase it for smaller particles Click the button Note As the settling time is sample dependant it is the operators responsibility to ensure that any remaining airborne particles are minimized according to the environment in which the instrument is situated and local health and safety regulations Page 6 11 Chapter 6 Page 6 le The Illumination dialogue is shown SOP Editor New SOP File Edit Help ae Oi Blk wi Sample details Pre measurement settings Measurement control Sample carrier Sample Dispersion Unit Illumination Optics selection Diascopic bottom light Analysis settings Filters Classification Post measurement settings Reports Calibration intensity 60 00 Intensity tolerance 0 20 Measurement tutorial Choose either Episcopic top light or Diascopic bottom light as the light source If using the optional filter plates Episcopic should be selected If Episcopic is selected the Calibration intensity and Intensity tolerance fields become active Enter a Calibration intensity suitable for the sample this intensity will be set bef
87. e Username this along with the user Password forms the unique key required to identify each individual using the system The Username is commonly an abbreviated form of the individual name or a unique identifier such as an employee code B Full Name the full printed name of the individual This can be used on reports to identify the user if an employee code is used as a Username B Description field optional this is used to add descriptive text for the user Note Q Only members of the Administrators group can add or edit user proper ties Toaddauser 1 Inthe Security Configuration dialogue select User New User or dou ble click an empty Username row to display the User properties dialogue User Properties User Must Change Password at Nest Logon C User Cannot Change Password 2 Type in the user information in the first three fields To set passwords the recommended approach is for the administrator to spec ify a previously arranged password such as the user s name and force the user to change their password the next time they log on by selecting the User Must Change Password at Next Logon check box Page 10 6 MAM 0410 Security Chapter 10 3 Press the Groups button to display the Group Memberships dialogue Group Memberships User Stuart Coombe Member of Mat member of AGE Administrators git Operators 4 Use the Add button to allocate the user to appropriate group s by moving each
88. e filec ge me ui gt lt Image of type BMP files bmp Image details Pixel size Microns per pixel Size W x H pixels 01 000 520 x 390 T BPP 24 File name Sample 1 Bmp If multiple files are selected the results from these are merged Highlighting a file in the file list displays it in the Image details viewer for quick visual verification Only the file s transferred to the Image files to analyse list at the right hand side of the dialogue will be analysed Page 7 4 MAM 0410 Measurement from files Chapter 7 Remove any unwanted files from the list using the button Use the Microns per pixel parameter to calibrate the system This is the length of the pixel side in microns The software assumes that pixels are square Depending on the original source of the image it should be possible to find the pixel size information microns per pixel edge The analysis can be run using the default 1 micron pixel but the particle size data will be incorrect Click the Q3 button 3 Inthe Sample details dialogue shown below type in the sample name and any notes associated with the measurement to help operators understand what the analysis does Image File Analysis Back o Image Files Threshold Sample name Analysis settings Filters Classification Sample 1 XYD Notes Comments Morphologi G3 Page 7 5 Chapter 7 Measurement from files 4 Click the G button The Threshold
89. e number of Additional layers below focus Additional layers above focus are the number of extra scans in addition to the scan taken at the point of focus for example if the number of Additional layers above focus was set to 2 the final Z stacked image would be the prod uct of 3 scans Sometimes the point of focus may not be at the surface of the plate for example when using the Wet Cell so it could also be useful record images below this point In such cases select a number of Additional layers below focus Refer to Chapter 2 for more information on Z stacking Note Running an SOP with Z stacking enabled takes longer as more images are scanned and compiled Click the button Morphologi G3 Page 6 15 Chapter 6 Page 6 16 SOP Editor New SOP File Edit Help Qe O H Ala Al ae Sample details Pre measurement settings Measurement control Background separation Sample carrier Sample Dispersion Unit Illumination Optics selection Sx 6 5um 420um Scan areas S 10x 3 5um 210um Threshold Scan areas Analysis settings Filters Classification Post measurement settings Reports Threshold 0 Due o EENEKREKWESy wJ 64 Measurement tutorial 12 The Threshold dialogue for the objective is displayed Source view 10095 o A grasse 128 192 255 Click the Get New Image button to capture an image of the current sample This opens the Microscope Manager so the user can move to
90. e parts of the dispersion chamber are shown below ill 8124 These are D Chamber body The circular sample cartridge is placed in a recess in the top of this Page 4 2 MAM 0410 Preparing the sample Chapter 4 Air tight seal on base forms a seal against the glass plate Screw on cap holds the sample cartridge in place keeping the sample air tight Air pipe connector this is where the air supply fits If the air pipe cannot be inserted easily check that the slider is pushed inward not out 9 Preparing and loading a G3S sample This section describes how to load a sample into the SDU The procedure is writ ten for a disposable sample cartridge The disposable sample cartridge comes in two parts an upper and a lower Each part is pre assembled with a foil disc already inserted Toload a sample for dispersion disposable sample cartridge Morphologi G3 1 2 Place the upper and lower parts of the sample cartridge on the bench Use the spoon to scoop up the sample Load until the hole is just over full then scrape away the excess using a scalpel blade for example Use the spoon to load sample into the centre of the lower sample cartridge The lower sample cartridge has deeper holes for the sample than the upper part of the cartridge ill 8134 Page 4 3 Chapter 4 Preparing the sample 4 Press the two halves of the sample cartridge together as shown be
91. e procedure the microscope is put into colour mode This is only used for image capture it has no effect on SOPs thresholds Scattergram tab etc Note It is not possible to make measurements while in colour mode The instru ment automatically switches back to monochrome Depending on the image it may be necessary to use the top light We recommend starting with bottom light and only using top light if the bottom light result is unsatisfactory For example with large spheres the bottom light may simply show silhouettes and not pick up their colour To capture the image 1 Inthe Microscope Manager adjust the intensity using the Light calibration button On a Morphologi G2 use the command Instrument Manual lamp control to enable the front panel Lamp dial then use that 2 Select the command Instrument Colour mode Wait a few seconds for the message Changing video mode to disappear then a few seconds more for the image to appear in colour The image may not look the correct colour at this point 3 Move to a suitable area ready for white balancing W For the bottom light move to the light calibration position m For the top light use a reference white area If necessary this can be placed on the slide White balancing is required because changing the brightness of the lamp changes its temperature and this changes the colour As the brightness is reduced the red part of the RGB mix becomes dominant white balanci
92. ean 1 Lactose 18 September 2007 16 39 24 False 23542 24 36 0 821 0 700 0 300 0 971 2 New Sample 18 September 2007 17 02 40 False 1187 28 50 0 637 0 493 0 507 0 952 3 Sample 2 03 October 2007 10 32 38 False 2509 30 01 0 668 0 518 0 482 0 951 4 Sample 3 03 October 2007 11 52 13 False 21178 30 65 0 654 0 502 0 498 0 94 03 October 2007 15 00 54 4903 30 92 0 645 FEE 0 501 0 944 Mean 5 Std Dev RSD Minimum Maximum i mdyson An example set of records in a measurement file named Example Results vmes is opened when the software 1s first launched after installation Subsequently the software opens the last measurement file that was used before shutdown Measure ment results are automatically saved to a measurement file To open a different measurement file select File Open Measurement Multiple measurement files can be opened if required The Records tab shows the records in the file It is the tab where all results or records and reports can be viewed moved and analysed The other tabs are WB Scattergram tab displays size distributions a scattergram class and filter information and particle images B Comparison tab used to compare multiple records B Report tabs there are a variety of these reporting the results Examples include the CE Diam diameter and HS Circularity reports shown in the above example To move to a different tab simply click on it Page 5 e MAM 0410 Software features Chapte
93. ed for example Filtered out Circularity gt 0 8 Return to the Records tab to see the data for the new record The original record remains available in its unaltered state If the button is not used on return to Records tab this dialogue appears Morphologi There are new particles selected For exclusion Do you wish bo generate a new result that excludes the selected particles Click Yes to produce a new record The Documentation dialogue then appears as described in step 1 Page 8 9 Chapter 8 Viewing the results Defining classes of particle Classes are described in Chapter 2 To set up a class using the scattergram 1 Select the Classification tab This will initially show no classes unless the SOP defined any Selection Classification Filters Exclude unclassified particles 1341 Parameter Operator Value 1 Value 2 Particles available for classification 1341 2 Inthe scattergram draw a box around the required particles 3 The Classification tab shows the parameter criteria like this Selection Classification Filters Exclude unclassified particles 527 Parameter Operator Value 1 Value 2 E Class 2 614 particles Exclude class CE Diameter pm Between 7 66 251 11 Area une Between 76 11 1574 05 select gt Particles available for classification 1341 Particles classified ol 4 4 If required change the values by re dragging the box or by typing value
94. ed par ticles only use the Classification list Page 13 11 Chapter 13 Creating custom reports This example shows particles in a class named Acceptable sorted by area in descending order Classification Acceptable gt Sorted by Area Pixels Descending wea T46 BFS 468 H 20 um Scale 1 1000 The View and Font tabs allow rescaling disabling the header display and font changes On the View tab the Show filtered particles check box controls display of these particles Graphs lin One or more Trend graphs Distribution graphs Classification graphs or Scattergrams can be added to a report Click on the graph icon and drag a mar quee on to the report B Trend graphs allow the measurement data from multiple records to be compared to investigate any trends in the information Here is an example Trend Graph TE T im c n o 3 Record Any parameter can be chosen for the X axis and another two numerical param eters for the Y1 and Y2 axes Page 13 le MAM 0410 Creating custom reports Chapter 13 WB Distribution graphs histograms frequency curves and oversize undersize curves can be displayed as well as combinations of these Here 1s an example CE Diameter um 1 100 CE Diameter Lim Recard 31 Sample x2 WB Classification graphs shows the number or percentage of particles in each class and how many unclassified particles there are Here is an example
95. een both frame parts A locating node is provided to ensure that the lower frame is inserted correctly The frame clamps together using a magnetic clo sure the whole unit should feel tightly clamped with no loose parts Screw on both Luer fitting caps to the Luer fittings Sample insertion 1 Morphologi G3 Ensure the volume of the sample mixture measures slightly more than the vol ume of the Wet Cell 2 5ml The volume of the cell can be increased by adding more gaskets Remove the caps from both Luer fittings Page D 7 Appendix DO Optional sample plates 3 Insert a syringe threaded syringes will screw into place into either of the Luer fittings and slowly pump in the sample Fill the glass cell slowly and smoothly until the excess liquid just starts to appear in the other Luer fitting ill 8217 Note Q Slightly tilt the Wet Cell so that the glass cell fills from the bottom upwards to prevent bubbles forming If the sample accumulates air bubbles during the filling process it is sometimes possible to eliminate them by removing one of the Luer fitting caps tilting the cell so that the open fitting is upper most and then lightly tapping the glass area 4 Remove the syringe replace both Luer fitting caps 5 Invert the whole wet cell so that the upper frame 9 is topmost and insert into the instrument Disassembly and cleaning 1 Hold the cell in the inverted position with the lower frame
96. efine classes On the basis of these selections new records can be created with some particles excluded The Comparison tab describes this tab and shows how to use it to compare multiple records The Report tabs lists the Malvern default reports and explains how to zoom in on reports identify data points on graphs and print a report Modifying graph formats explains how to change the appearance of graphs in reports Interpreting classes in the display explains the meaning of the Classification Chart display Capturing colour images of particles shows how to capture coloured images Page 8 1 Chapter 8 Viewing the results Records tab When the measurement is complete the new measurement record appears in the Records tab By default records appear in the order that they were measured 77 Morphologi Example 2 vmes 3 J Eile Edit View Measure Tools Security Window Help Das amp ld i Malvern Default i Dom G amp I Comparison i ceEDiam m fi cEDiam von m HS circularity M E Aspect Ratio M Record Sample Name SOP Name Date Edited Particles CE Diameter Mean um HS Circularity Mean Aspect Ratio Mean E 1 Salt Glass Beads 5 2 5x 09 November 2006 10 06 04 False 1080 58 53 0 927 0 943 3 Salt Glass Beads 2 5 2 5x 09 November 2006 10 06 04 True 1074 58 23 0 327 0 343 4 Salt minus Glass Beads EE 5 Sakminusoblates BE E RC NN RR REEL RR 1 6 Salt minus oblates EFETEEEEIICITTZCTECU
97. ent Column Title Argument Value E 2g Details Record Image DB Sample Name E fe Measurement Audit Date a Measurement Properties Particles a Morphology CE Diameter Mean um iE a SOP Settings CE Diameter RSD Type CE Diameter D n 10 um x 0 1 CE Diameter D n 50 um x 0 5 CE Diameter D n 90 um x 0 9 HS Circularity Mean Convexity Mean Elongation Mean Length Mean um Width Mean iim Y 4 Use the Settings tab to specify how to format the data The parameters are Include header row v Inserts headers into the spreadsheet Enclose text in Encloses each parameter separately quotation marks Use tabs as sepa Spreadsheet packages require different field separa rators tion characters to space the data output correctly Use commas as Use tabs to output to Microsoft Excel separators Commas are most common but do not use these if they are used as numerical separators in the local ised Windows version for example the French ver sion of Windows 5 Select the Parameters tab and build the list of parameters to include in the exported data The list on the left displays all available parameters The list on the right displays all parameters that are to be included in the export template Add parameters to the right hand list or remove them from it using the gt and buttons Page 9 9 Chapter 9 Page 93 10 Exporting results 6 Ifa parameter requires an argument it ap
98. eparing the sample gt To use evaporative dispersion 1 Place an aliquot of approximately 2 mg of sample depending on the particle size in a sample vial 2 Add around 4 ml of the solvent system 3 Depending on how fragile the particles are or how agglomerated apply vortex mixing and or ultrasonication to the sample suspension 4 Cover a clean glass slide with approximately 0 5 ml of the sample suspension as shown below Ensure that the entire slide is covered by the suspension ill 8138 During this procedure it is important to m Keep the slide level B Support the slide above the bench so that air can circulate around it Do not place the slide on a solid surface B If available use a stand which has parallel rods to support the slide above the bench and feet which can be adjusted to keep the slide level This is shown in the above diagram 5 Place some sort of cover over the slide as shown below B to slow evaporation preventing pooling and contamination Page 4 8 MAM 0410 Preparing the sample Chapter 4 ill 8139 6 Leave the solvent to evaporate Experience shows that slower evaporation results in less pooling of the sample 7 Once dry the slide can be measured Wet dispersion wet measurements Oil dispersion This means dispersing the sample in oil It is particularly good for monodisperse samples During method development several trials may be required to determine the most ap
99. erical aperture 0 040 0 075 0 15 0 30 0 40 0 55 Focal depth total um 343 75 97 78 24 44 6 11 3 44 1 82 Working distance mm 3 2 8 80 18 00 15 0 13 0 9 80 Camera system Camera type 1 Global shutter progressive scan colour CCD Connection protocol type IEEE 1394a FireWire Number of pixels 2592 x 1944 5 MegaPixel Pixel size 2 um x 2 um Sensor size 7 15mm x 5 44mm Slide holders SDU glass plate 180 x 110mm G2 four slide plate holds up to four 76x26mm microscope slides G2 glass plate 100 x 120mm MAM 0410 Specification Appendix A Minimum computer specification Contact the Malvern Helpdesk or website for the recommended computer specifi cation otherwise consult the Software update notification document supplied on the software CD The computer is supplied with the system The main compo nents are Computer B Mouse and keyboard B Two 17 flat panel monitors one for software and one for live video feed Chemical compatibility Morphologi G3 Components of the Morphologi G3 that may come into contact with the sample are manufactured from materials that are considered to give the widest protection from chemical attack However it is important to check that any sample used is chemically compatible with the materials mentioned Page A 3 Appendix A Specification Page A 4 MAM 0410 Unpacking instructions Appendix B of the Morphologi G3 User Manual MA
100. es between 3 5um and 6 5um 20x 1 8m 100um 5x 6 5um 420m 2 5x 13um 1000jiIm Morphologi G3 Page 6 13 Chapter 6 Measurement tutorial 10 If multiple objectives are selected the following Objective Threshold and Scan areas dialogues will be repeated for each set Click the button 11 For the first objective selected above the Objective dialogue is displayed Its title bar shows which objective it is for and the relevant particle sizes SOP Editor New SOP File Edit Help Qe Ql Sample details Pre measurement settings Measurement control Sample carrier Sample Dispersion Unit Illumination Optics selection 50x 0 5m 40pm Threshold Scan areas Analysis settings Filters Classification Post measurement settings Reports Measurement Overlap 40 00 Facus Manual Focus O Fixed Focus Offset um Select Z stacking mode No 2 stacking Additional layers above focus Additional layers below Focus Total Z stack range The Overlap can be modified to extend the range of a given objective but should normally be left at the default 50 for the 1X or 2 5X 40 for the other objectives This is described in Chapter 2 Note Q There is a trade off as a larger overlap means a longer analysis time If required select Enable particle stitching as an alternative method to Overlap See Chapter 2 for more details on particle stitching Note This option is only available on
101. eshold 0 64 126 192 259 0 Gi lH 247 Drag the i button to slide the Threshold bar to change the applied threshold level Aim for a threshold level which correctly identifies the particle edges This is the point where a light grey halo starts to appear around each particle The two numbers and define the range selected To contract the range itself use both the and buttons Page 6 17 Chapter 6 Measurement tutorial High contrast darker particles normally need a lower threshold lighter crystal line samples generally need a higher threshold The graded scale under the slide bar is a reminder of this The default setting picks up the darker parts of the image Optionally click the Invert checkbox for a negative view of the standard threshold image This can be helpful when trying to refine an appro priate threshold setting and 1s particularly useful where the background is darker than the particles Where both light and dark particles co exist on the sample the dark and light threshold level sliders can be used to contract the range in order to segment both particle types A standard Threshold calibration using just the 4 amp 8 slider has been set in the illustration below Background separation Source view 4058 Using both 4 and buttons together the two lighter particles are excluded Packer nana oro sica Gourde view HFA amp Jul screen Get new mane E
102. existing SOP and saving it with a different name To modify an SOP 1 Select File Open SOP or Measure SOP Settings The SOP Editor dialogues described in Chapter 6 will be displayed Change any settings as required 2 Select File Save As and specify a name for the new SOP Alternatively just click OK to replace the original SOP with the modified one If measuring similar samples it is often easier to modify an existing SOP and then save it under a different name The Filter and Class SOP Editor dialogues have Copy and Paste buttons which allow quick copying of filters and classes between SOPs These can also be pasted from the Scattergram tab Distributing an SOP SOPs are designed to allow the measurement of similar sample types in a consistent way When monitoring batches of sample in a Quality Control environment the results will be meaningless if different measurement parameters have been chosen for each batch Consider an application where a manufacturer is producing the same type of sam ple in different factories It will be important that the measurement protocol is con sistent between factories An SOP can be created and copied into the SOP directory on another Morphologi G3 instrument so that measurements made on the two instruments are consistent Simply copy the vsop file to all computers running a Morphologi G3 system The default location for SOP files is C Documents and Settings All Users Shared Doc
103. fill the whole window B Video the Live picture window expands to fill the whole window as shown in section O above The Microscope Manager The Microscope Manager window shown below gives the user control of the microscope It opens in several modes i W For manual control opened using Tools Microscope or the button B From the SOP Editor when accessed from the SOP Editor s Threshold dialogue it s used simply to select an image for analysis B From the Scattergram tab when the user selects a particle then View in original image When opened for manual use the window is similar to the following Fl Microscope Image Instrument Joystick Help Glass Plate 120x100mm Slide 1 E Z Position Control Step Size 69 07 um 80 Custom light calibration c iN Standard light calibration zZ Stacking 2 es Perform Z Stack x Y Position 2 Position 35 373 mm 81 575 mm 0 255mm 3 Move to preset position v Fit to window L 200 um J Ca Page 5 20 MAM 0410 Software features Chapter 5 Toolbar Use the two pull down lists to select the plate and for a two four slide plate or the filter plate the individual slide The buttons are shown below Button Function Moves to the light calibration position Moves to the plate or slide centre B e Toggles between diascopic bottom light and episcopic top light When a particle is selected in the Scattergram tab this butto
104. ful for making an analysis comparison between two sam ples sequentially In addition to general use it can be used in pre existing workflows that utilise the glass sandwich approach to presenting filters Each slide is held in place by a sprung clamp Part No Description MOR5320 Two slide plate The Two Slide Plate is shown below ill 8215 Usage Slide insertion 1 Ensure that the Two Slide Plate is clean before inserting slides 2 Hold open the sprung clamp and insert a slide so that it rests on the slide plate 3 Release the sprung clamp to lock the slide in position Load the other slide area as required Pege D 10 MAM 0410 Optional sample plates Morphologi G3 SOP configuration Appendix D B Select the 2 Slide Plate 75x50mm in the SOP s Sample carrier section SOP Editor New SOP File Edit Help Qe 9g Sample details Pre measurement settings Measurement control Sample carrier Sample Dispersion Unit Illumination Optics selection Analysis settings Filters Classification Post measurement settings Reports Carrier Select the type of plate that will be used 2 Slide Plate 75x50mm Options Compensate For plate tilt C Coverslip over sample In the Optics selection Scan areas section of the SOP the system selects a rectangular scan area of the correct size for the entire slide this can be reduced if required Page D 11 Appendix DO Page D le O
105. g For example a user might be comparing two sets of measurements from different proc ess lines that are known to have different size distributions but want to know if the shape distributions cluster according to the process line To check this they can label the records as belonging to one of two groups When they select these grouped records any parameters that do not cluster according to the grouping will show the variability bar in grey The coloured bars indicate parameters that have a clustering pattern that matches the grouping pattern Page 8 15 Chapter 8 Page 8 16 Viewing the results The results The results comprise five displays for the parameter selected in the Parameter Variability list These are Parameter Variability The Parameter Variability bar charts alongside parameters show size green shape red and intensity blue parameters The radio button selected initially shows the morphological parameter which is responsible for the most variability between the selected records Its name is also shown in bold text The width of the bar shows how much variability between the records the parameter produces If groups are selected the bars which cannot group correctly appear grey If the user s defined groups are viable the bars remain coloured Dendrogram The links on the dendrogram cluster the most similar records together according to the parameter selected under Parameter Variability The clustering
106. gation 1 Elongation values range from 0 to 1 A rod for example has a high Elongation Morphologi G3 Page e 9 Chapter 2 What is the Morphologi G3 m Max Distance this is the furthest distance D between any two points of the particle Max Distance which is also known as the maximal Feret diameter or caliper length is reported in um ill 7690 B Perimeter the total length of the object boundary in um calculated by sum ming the length of the boundary pixels This includes an adjustment to take account of direction changes The perimeter below is amp O A ill 7698 m CE Diameter the diameter D ofa circle with the same area as the pro jected area of the particle image shown below ye Here Particle area Circle area 9 CE Diameter is reported in um ill 7689 B Circularity the ratio of the circumference of a circle equal to the object s projected area to the perimeter of the object This is calculated as 2 x In x Area Circularity Perimeter Page 2 10 MAM 0410 What is the Morphologi G3 Chapter 2e Morphologi G3 As described earlier in this chapter a perfect circle has Circularity of 1 0 while a very narrow elongated object has a Circularity close to 0 HS High Sensitivity Circularity the ratio of the object s projected area to the square of the perimeter of the object This is calculated as HS Circularity 4 x n x Area
107. hat can be emailed to other users Note Q To create an SOP for an image file rather than a sample see Chapter 7 Measuring the sample the software selects the magnification and calibrates against a grating We recommend using fixed focus for extracted SOPs the user has to set the initial focus but otherwise the user focuses with the soft ware and then starts the measurement Viewing editing results viewing information on each individual particle or statistics for the whole distribution The user can use the Scattergram tab to define filters or classes sort and filter particles and create new records with cer tain particles excluded The Comparison tab allows easy comparison of records based on morphological parameters Full details on this are given in Chapter 8 Page 6 1 Chapter 6 Measurement tutorial 5 Creating reports displaying histograms of the distribution and result statis tics full details on this are given in Chapter 8 The Report Designer can be used to customise the style and content of reports see Chapter 13 for details Getting started The first steps involve powering on the instrument and starting the software gt To power on the instrument 1 Turn on the microscope at its power switch 2 Leave the instrument powered on for five minutes before any measurement is made This allows it to stabilise gt To start the Morphologi software 1 Double click on the Morphologi fa deskto
108. he form of sliders which are pushed into the microscope s optical path are designed to be used together These options are purely for manual microscope use they are not for use when using an SOP Although the options cannot be used for running full measure ments they can be used to obtain more information on particles of special interest The resulting image can be saved as a file and this then measured to obtain parame ters for the particle Page 3 15 Chapter 3 Page 3 16 Hardware features Caution Use of these sliders for diascopic microscopy is not recommended as this means placing the Polariser slider near the microscope lamp house and this may damage the slider To insert the Polariser and Analyser remove the triangular panel from the right hand side of the instrument as shown below ANALYZER K POLARIZER ill 8137 This reveals the two labelled slots for the sliders The triangular cover panel can be stored in the triangular recess below the LED panel on the left of the instrument The Polariser and Analyser look like this ill 7705 MAM 0410 Hardware features Chapter 3 The components are Adjustment screw This is marked P on the Polariser and A on the Analyser Polariser or Analyser Empty hole for when option is not in use To use the Polariser 1 3 4 Remove the rubber cap from the POLARIZER slot on the right hand side
109. hemselves precisely to the edges of the background grid For less rigid alignment hold down the ALT key whilst dragging an element 5 Right click on the element and use the menu this displays to configure its properties as described below Page 13 6 MAM 0410 Creating custom reports Chapter 13 Setting up the report elements Set the properties of any added element by selecting it and double clicking to open the Properties dialogue box The online help in these dialogues details how to use each one The sections below show the properties each type of element has Note Q This dialogue can also be opened using Edit Properties by right clicking and selecting Properties or by typing Alt enter on the keyboard Text A Use the Text tool to add general text such as a report name to the report With the element selected use its Properties dialogue to edit the text and its style including its alignment colour and size Edit text or type new text using the Cap tion box on the General tab Picture i The Picture tool is generally used for company logos etc All the main graphics file formats are supported With the element selected use its Properties dialogue to browse for the picture to insert Use the Keep Aspect ratio check box if required to maintain the aspect ratio of the original image Use the Link to file check box to link to rather than embed the file this means that when the file is updated the contents of the
110. hich are measured dry Users can of course also use their own methods Whichever technique is deemed appropriate for a specific sample during method development do test the reproducibility and robustness of the method Dry dispersion dry measurements Manual dispersion free flowing powders If the particles in the sample are large and free flowing consider manual dispersion by simply sprinkling the sample onto a large glass plate For this option the 1X Adapter Kit can be used with its walled plate and sample retainer see Chapter 3 Wet dispersion dry measurements Evaporative dispersion For some samples dispersion by an evaporation technique is appropriate This is particularly useful for very fine particles where a sample preperation device may cause static or breakage or polydisperse samples which make wet methods difficult The sample must not be soluble in the solvent used it is often best to add a sur factant to the solvent system to prevent agglomeration of particles A typical solvent system is freshly prepared 0 01 lecithin in iso octane W V Combinations of other organic solvents and surfactants such as Tween or Span can also be used for this technique as can a low boiling point silicone oil such as hexamethyldisilonane During method development adjust the concentration of the sample in the solvent system and the ultrasonication applied until a reproducible dispersion is obtained Page 4 7 Chapter 4 Pr
111. his inserts a border around an area B Parameter any parameter stored in a record can be added to the report B Calculation custom calculations using system parameters These can be used to provide pass fail decisions B Particle image grid used to show images of selected particles B Graph data in a selected record can be added to the report in the form of a trend graph distribution graph classification graph or scattergram B Classification table a table summarising any classes used These are added by using the Toolbox Page 13 2 MAM 0410 Creating custom reports Chapter 13 Note During creation of a report data from the current record s 1s used If no record is selected dummy data is used Getting started To open the Report Designer select Tools Report Designer The Report Designer opens in a separate window as shown below In this example the Toolbox has been dragged from the docked position to make it easier to use This tool can be used in either position depending on the user s pref erence E Report Designer VisionReportPage1 Screen Layout File Edit View Layout Configure Tools Window Help Ose b a EB cS i 10095 Parameter Calculation Frame Eit Particle Image Grid Distribution Graph Trend Graph Classification Graph EEE Scattergram i Tables Classification Table mer Press F1 These Edit menu commands are of general use WB Edit Copy and Paste a
112. ialogues appear showing the original measurement settings and any post analysis filters or classifications that were applied to the result These can then be saved as an SOP so measurements can be made again with the same settings This is useful to find a fixed focus value or if the parame ters of the measurement record are not already saved in the SOP directory MAM 0410 Software features View menu Chapter 5 Use the View menu to configure the main window and to create edit workspaces The options are Command Toolbars Function Selects which toolbars will be shown in the main window Only toolbars shown with a tick are displayed The Customize sub option allows further customisation of the user interface Status bar Toggles on off the display of the status bar Statistics bar Controls the display of the statistics bar Workspace Selects the measurement workspace to be used The three sub options are for adding editing or deleting workspaces For editing available workspaces are listed Records Scattergram Comparison lt Report name gt Measure menu Switches between the available tabs Click on a tab to dis play it Select this menu when ready to perform a measurement Once the measurement details have been entered or an SOP has been chosen the Measurement Man ager will appear Command SOP Function Opens an existing measurement SOP Image file
113. ied particles and fil tered particles too Data for all classes 1s exported to the same file m Settings exactly as described in the previous section For easy interpretation of the exported data specify Use tabs as separators and use a spreadsheet package to view and process the result Output example The following example shows the start of a comma separated file which includes a header row Id Magnification CE Diameter um Length um Width um Max Distance um Perimeter um Major Axis Class Name 19376 L 00 2954055 3392 02 298584535 11 109 12 150 84 1mDUTlUty ise 9oB aL 00 41 9044 LI 148754349949 9 441 158 5 1 08 Impa o Ves ao This file was produced using the Export particles in classes option on the Clas sification tab All particles in the first class Impurity in the above example are listed then those in each successive class Exporting images Morphologi G3 Images of each particle in a single record can be exported as bitmap bmp files This option is not available if multiple records are highlighted Select File Export Images and specify where to create the files Note This can create a large number of large files if there are many particles in the record The folder containing the images will resemble this after the export mak r File Edit View Favorites Tools Help Qo Back d A Search lg Folders X e M Address C TEMP Images b Go Mame Size Type Date Mo
114. ift keys Statistics bar The statistics bar at the base of the window displays the following for the selected records 1 and 3 in the above example B Mean the number s after the word Mean show the records selected This is the mean of the means for the selected records B Std Dev the Standard Deviation B RSD the Relative Standard Deviation B Minimum and maximum values The statistics bar updates automatically when new records are selected To hide any statistic use View Statistics bar or right click on the bar Page 5 3 Chapter 5 Software features Fixed columns Columns shaded in grey like columns 1 and 2 below are termed fixed columns These cannot be reordered by clicking on them They are fixed in the horizontal direction and remain visible even if the user scrolls over to the window s right side E Records Record Sample Name 5 Comparison Classification chart M SOP Name Date Edited Particles 1 Salt Glass Beads REF 09 November 2006 10 06 04 3 Salt Glass Beads 2 S 2 54 09 November 2006 10 06 04 True 1074 5 Salt minus oblates 2 5x 09 November 2006 10 06 04 True 119 6 Salt minus oblates S 2 5 03 November 2006 10 06 04 True 990 7 Salt minus oblates 3 5 2 5x 03 November 2006 10 06 04 True 990 Scattergram tab The Scattergram tab displays size distributions class and filter information and particle images Use of this tab is described in detai
115. in WING OW 3t 9 acad Y P dH ERI RR CUR ECCE NC ERO Rr ec 5 2 Sotware COMPONEMIS 4 43 4 doe qa ans PR dca e Ca ea ve c x e Re rot 5 8 The Measurement Manager leen 5 16 The Microscope Manager eee eee eee rn 5 20 B WelMjele l m ER REEL t ea eae we ae Se aes 5 25 Backing up and Archiving the system llle 5 26 Measurement tutorial Introd CLOS esana ed dC oe e a DECIES a af ded deme et e DO aa 6 1 Ge tlilig States 20s Bcd ato hates Gode Meade e Road EC ee ae dead 6 2 T Method development 2 5 qaa 940 03 aac caca CA d E US AR OO e E 6 2 2 Creating amo Ps Shanda hon eee dts a ae rA CIRCO o FR CA CR RE aed Je E Sor n 6 6 3 Measuring the sample seen 6 28 4 Viewing and editing results llle 6 36 5 GreatlHd repons 4 562 9 1 3 2728 rd La e OR RECO oe ae ee Pea Es 6 36 Measurement from files NOC COM A ERE ta dg sya es ee ee D 5 12572 012 7 2T TA 7 1 Getting stated eae i os ton Retreat ana PAD ce Mead at a a Soa 7 2 The SOP Edo ragi 4 23 Cb dC uee en r do peritos d is 7 3 Measuring an image file nnana ce es 1 4 Viewing the results IuingoroDrenje PRICE 8 1 Records tabia d aac EE ae SUE Sad ad CAL obsit aiu tacit ed 9 2 Thesoattergrami tabs 2233 2 cR ee hts e EC eee ee EES 8 4 The Comparison tabs ax dd a3 a RR DO Onde RACIO Bede ee UR a 8 14 REDON 18S 4 4 6 456 bk hee ees a Sawa mag Oh OSD wes 8 18 Modifying graph formats 2 0c ee hn 8 21 Interpreting classes in the dis
116. in the Records tab When all parameters are set up switch to the Report Pages tab and proceed as described below or click OK to exit and return to the Records tab The new workspace will become the current workspace Selecting reports Use the Report Pages tab to specify which reports either created by Malvern Instruments or user defined to include in the workspace The workspace will include a tab for each report type selected here gt To select reports to view Morphologi G3 Select the Report Pages tab All available reports are listed as shown below this example does not include any user defined reports ELLE LE Tm Record View Parameters Report Pages Select those reports you wich to display and the order in which they appear v EE Diam Patticles M FRAIHS Circularity M amp rea pm M amp spect Ratio M CCE Diam M CE Diam val M CE Diam Circ Classification M CE Diam HS Circ M Cire Particles M Cireularity M Classification chart M Classification table M Convesity M Convexity HS Cire M Elongation M J Histogram Undersize Distribution M Page 12 5 Chapter 12 Workspace management 2 Select the check box for each required report 3 If necessary use the UP and DN buttons to change the order the report tabs will appear in this is initially alphabetical Click OK Any reports created with the Report
117. is hierar chical records are clustered into groups then groups clustered into bigger groups In the example below grouping records 4 and 5 and records 1 3 and 6 as two clus ters groups minimises within group variability for two groups on the selected parameter Elongation Dendrogram Rl Sal Glass Beads R3 Ssaltt Glass Beads 2 Re Salt minus ablates RS Sat minus oblates R4 Sal minus Glass Beads I I L I I I I I I I r I I I I I I EF I I I I I I L I I I I I I r I I Q 1 rE 4 a rR 2 0 4 0 6 0 8 1 Record Differences Elongation A warning icon and warning message tell the user if the selected records are not suitable for clustering The X axis indicates the difference between two records or two groups or the dif ference between one record and one group of records These differences always MAM 0410 Viewing the results Morphologi G3 Chapter 8 range between zero and one Zero means there is no difference between two records One means the two compared records groups have a completely different range of morphological values In other words their distribution curves do not overlap at all Trend plot The trend plot shows the mean value of the parameter selected under Parameter Variability for each record Comparisons are based on distributions so do not provide a single parameter to monitor The trend plot though shows the mea
118. is is useful for setting Process Control thresholds Frequency and Undersize curves initially the morphological parameter that shows the biggest difference value between two records is selected for showing these two distribution plots If the user selects a different parameter in MAM 0410 Software features Chapter 5 the curves for that are displayed The Undersize Curve is also known as a cumulative oversize curve or result under plot Report tabs A Report tab shows either a default report format provided by Malvern Instru ments or a user designed custom report produced using the Report Designer as described in Chapter 13 A report can show a graphical summary of the distribu tion or of one or more morphological parameters It can include individual particle images Most reports will eventually be printed to paper to summarise results etc The available Malvern Instruments reports are listed in Chapter 8 Chapter 12 shows which reports are available in each of the supplied workspaces The follow ing example is a Malvern Instruments report 5 Morphologi Example Results vmes Record 1 Salt Glass Beads File Edit view Measure Tools Security Window Help Daw SB gt Comparison en heen CE Diam Circ Classification M B CE Diam HS Circ 4 e CE Diam Particles M Morphologi Convexity amp HS Circularity Number Distribution Ios hs E Combined reports 7 E Convexity HS Circ M J
119. is scanned and digital images produced Images of each particle are saved providing visual verification of the analysis data The analysis software measures selected characteristics of each particle Page e l Chapter e Page e e What is the Morphologi G3 4 A Scattergram tab allows different types of particle to be classified easily according to their shape characteristics 5 A Comparison tab makes it easy to identify the key shape differences between samples 6 Reports including frequency distributions and histograms are generated These can be user defined reports if required 7 Saved data can be exported for use and further analysis elsewhere The Morphologi G3 was designed with simplicity in mind so minimal user inter action is necessary to achieve excellent results The use of SOPs and the statistical significance gained from analysing a large number of particles ensure that results are as objective and reproducible as possible What are particle size and shape This section is an overview of particle size and shape and why they are important What is particle size Describing a three dimensional 3D particle is often more complex than it first appears For practicality or management purposes it is convenient to describe parti cle size as a single number However unless the particle 1s a perfect sphere which is rare in real world samples there are many ways to describe its size This 1s the
120. it appeared initially Unless the particle was from an image file the microscope must be con nected Xim Manual exclusion tool click on this then click on particles to exclude them Individual particles can also be excluded included by right clicking and selecting Manually exclude particle or Manually include particle Use these options to include exclude particles Morphologi G3 Page 8 7 Chapter 8 Page 8 8 Viewing the results The value of the selected parameter for each particle is shown below it in the example below it s the Id and filtered particles are marked with a red X like par ticle 9 below JF Selected Particles Id Creating particle filters Filtering particles means excluding one or more particles from the selection based on the value of any morphological parameter Individual particles can be removed using the toolbar as described above More complex filters can be set up based on one or more morphological parameters Multiple filters can be set up and then combined Filtered particles appear in the display with a red X as shown for particle 9 in the above example Right clicking on a particle marked like this undoes the action of the filter s applied to that particle To set up filters 1 Select the parameter to filter on above the major filter graph 2 Select the Filters tab Any filters already defined in the SOP will be shown 3 Drag the left and or right margins inward t
121. it to the right hand list This example shows a list with several parameters Record View Parameters Report Pages m Area um Column Title Argument value ey Area Pixels Area Maximum pixels Poo Pe 4 Area pixels D v x Area Mean pixels Area pixels Number Distribu Pixel Area RSD 95 Pu Area pixels Volume Distribut El Pixel Area Din x Pixel Area Bands pixels t X 9 Pixel Area D n x Ect Statistics y Area pixels D 3 2 bis Area pixels D 4 3 ine Area Maximum pixels jen Area Mean pixels 9 Area Minimum pixels Area STDY pixels 4 Pixel Grea RSD 9 5 Cancel Page 12 4 MAM 0410 Workspace management Chapter le If an added parameter requires an argument to be supplied an x appears below it The parameter Pixel Area D n x shown above is an example Type in its value in the Argument Value column The example above has parameters of this type with argument values of 10 and 90 producing parameters for the 10 and 90 percentile values respectively Column names can be changed To change a column name double click in its Column Title field When the cursor appears in the field edit the name Parameters appear in the right hand list in the order in which they will appear in the Records tab To move a parameter select it and use the or but ton to move it up or down Parameters at the top of this list appear furthest left
122. k the amp button If more than one objective is selected the sequence of Optics selection Threshold and Scan area dialogues steps 10 to 12 is repeated for each objective Page 6 20 MAM 0410 Measurement tutorial Chapter 6 15 When all objectives have been set up the Analysis settings dialogue is dis played SOP Editor New SOP File Edit Help Qe O11 elh Az Sample details Pre measurement settings Measurement control Analysis ID Sample carrier Sample Dispersion Unit 3 00 Illumination Optics selection 5x 6 5um 4201m Trash size Threshold Minimum number of pixels Scan areas 10x 3 5um 210pm 10 Threshold Scan areas Analysis settings Segmentation method Filters Classification b Post measurement settings Reports Fill holes Enable hole Filling Size bands Specify new values for any of the following parameters B Analysis ID the default should be the highest number in the list Only select a lower number for backwards compatibility with data from older versions of the software For example 2 00 gives compatibility with data from Morphologi G2 releases prior to 6 0 and 1 00 gives compatibility with the PVS 830 a predecessor of the Morphologi G3 B Trash size discard any particles smaller than this size pixels Use this to reject small dirt particles as noise but do not set it too high or par ticles may be lost forever In general it is best to leave this figure low and filter
123. l Using tweezers carefully insert the filter holding the sample so that it rests on the filter recess The tweezer recess helps to enable the correct placement of the filter Close the filter clamp to lock the filter into place The filter should be stretched taught when the clamp is closed If there are any noticeable kinks in the filter open the clamp and re insert the paper If required load another fil ter in the other filter area Tip If the sample preparation area is not within immediate proximity of the Morphologi instrument the sample s can be covered with the filter protective cover This can prevent contaminants from entering the sample and the dis turbance of dry samples whilst in transit to the instrument Remove the cover before inserting the filter plate into the Morphologi instrument The Filter Plate can be washed in a dishwasher Page D 3 Appendix DO Optional sample plates SOP configuration B Select the Filter Plate 47mm or Filter Plate 25mm from the Sample carrier section of the SOP SOP Editor Mew 5S0p Be tdt Heb Om Qi Sarna det ads Pre meadgement sett Pol e e d umbra Reports m Inthe Optics selection Scan areas the system automatically selects a circular scan area Bear in mind the following maximum scanning areas when prepar ing the sample Filter plate Maximum scan area radius 25mm 10mm 47mm 21mm B During the running of the SOP the user is p
124. l in Chapter 8 Its components are shown below ord 1 Sali Glass Beadk perum iE Be Edt wee Mesure Took lecurty Window Help Z Morphologi Exarmple vmes R 2 I Fiki area Pics Feed unn 353 12 pex Rosa n 327 CE Dimeker um 2 02 Centre X Postion um Jp or eh Cente Y Foution ur Lrculanity Selecicn Claitfheaion Fikers e E Hee Elche unclassified particles 257 7 Firme Dm atir Vale 1 Value ft a inktensty 5D 3 Glace Spheres 075 partic Exchade clase o LU aia HS cule h E i E 5 Mace Ras E 26 Aspect Pata T na Zion e amp assai ide e ped Loris SE Volare jam TIS5 6B anksty dk um Bead ill 8068 The components are D Scattergram pane used to select the parameters used define filters quickly and select particles of interest It shows the distribution of particles on a plot of any two selected parameters Selection Classes and Filters pane used to make quick selections based on parameter values also create or modify classes and filters These can also be set up as part of an SOP as described in Chapter 6 Particles toolbar controls the particle display Page 5 4 MAM 0410 Software features Morphologi G3 Chapter 5 Particles pane shows the particle images The images initially appear in the order that they were detected i e based on their Id The selected parameter
125. l tab click the Select button In the Select a parameter dialogue select the appropriate parameter see below and click OK Use the Label selection area to add a parameter label alongside the parameter value The style colour and font of the label and value can also be defined here Parameters with arguments Certain parameters require specific values or arguments to be added After select ing one of these use the Parameter arguments box to enter values Parameter engen Column Title Argument Value E LE Diameter D n um To set the argument value double click on the Argument Value box and type the value in Next alter the Column title to reflect the argument value by double MAM 0410 Creating custom reports Chapter 13 Morphologi G3 clicking on it For the above example the user might enter 0 1 as the Argument value then change the Column title as shown below Parameter arguments Column Title Argument Value CE Diameter D n 10 um BE ae 11 Calculation E In addition to the defined parameters described in the parameters list custom cal culations can be entered This is useful for displaying Pass and Fail criteria for morphological parameters New derived parameters can also be created such as CE Diameter Elongation With the element selected use its Properties dia logue to configure the calculation Properties General Style Col
126. ld setting is too high and that some of the background is being misinterpreted as particles It may also prove useful to see how the threshold affects shape parameters or to use the scattergram tab to examine the captured particle images at the different thresh old settings If necessary further analyses can be performed on the stored frames at other threshold settings When a threshold setting that provides the optimum information has been estab lished edit the original SOP and set the threshold to the preferred setting Page E 3 Appendix E Optirnising threshold settings Page E 4 MAM 0410 Index Numerics 21 CER part 11 10 2 10 4 21 CFR part 11 icon 5 8 5 15 A Access to the instrument 1 3 Administrator setting up 10 3 Analyser 3 15 Analysis ID 6 21 7 8 Analysis settings dialogue 6 21 Aperture diaphragm lever 3 5 Appending results to file 9 4 Area pixels and Area microns 2 8 Argument in Export template 9 10 in report 13 8 Aspect Ratio parameter 2 9 Audit Trails 5 25 Axis settings graph 8 22 B Background separation 6 16 7 6 Batch printing 8 3 Brighter button 5 21 C Calculation Editor 13 10 Calculation in report 13 9 Calibration algorithm 6 34 Calibration gratings 3 11 Caliper length 2 10 Camera components 3 13 Capturing colour images 8 27 CE Diameter 2 10 CE diameter 2 2 Centre X or Y position 2 7 Choosing magnification 6 5 Circle Equivalent CE diameter 2 2 Circularity 2 4 2
127. ld text and shown in the form main menu menu item submenu item For example the command File New SOP refers to selecting the SOP submenu item under New in the File menu Where to get help This section describes the available sources of information on the system Manuals and online help The primary sources of information on the Morphologi G3 system are the Quick start manual this manual the Essentials manual and the software s online help The manual gives an overview of the system as a whole while the online help sys tem gives detailed information on the Morphologi software Each Morphologi soft ware dialogue has a Help button giving information specific to it Quickstart manual This gives a simple introduction to the system and explains how to make a meas urement MAM 0410 Introduction to this manual Chapter 1 Morphologi G3 Essentials manual The Essentials manual covers Site requirements all the physical requirements for positioning the system Information is given on service requirements air number of power sockets etc environmental requirements temperature humidity etc and physical requirements space needed etc Note The system must be initially commissioned by a Malvern trained repre sentative Health and Safety this must be read by all users of the system It details all safety issues for the instrument and samples Maintenance this includes characterisation and tro
128. le Browse for it or type its name in the File Options selection box Specity whether to B Append to file add to the end of the existing file B Overwrite file replace any previous content in the file Page 9 4 MAM 0410 Exporting results Morphologi G3 Chapter 9 Parameters tab Use the Parameters tab to export selected parameters only Export Data File Parameters Settings Export template Q Use export template parameters Export temp 1 The options are Use record view parameters exports the parameters currently displayed in the Records tab These are defined by the workspace Use export template parameters exports parameters defined by a tem plate Available templates will be listed below select one and click OK To cre ate a new template or edit an existing one see Creating an export template later in this chapter Settings tab Use the Settings tab to specify how the data is exported It has these options Include header row inserts the parameter names as the first row in the spreadsheet Enclose text in quotation marks encloses each parameter separately Use tabs as separators Use commas as separators spreadsheet packages require different field separation characters to correctly space the data output Commas are most common but do not use these if they are used as numerical separators in the localised Windows version for example the French version of Windows For
129. llow copying and pasting of text and graphics from other programs The dh toolbar buttons have the same function m Undo 2 and Redo 5 5 for undoing or repeating commands respec tively Morphologi G3 Page 13 3 Chapter 13 Creating custom reports Opening an existing report To open an existing report use the Report Designer s menu command File Open Report files have the extension vrep Creating a new report A new report can be created in two ways B Selecting File New from a blank page and adding the report contents B Opening an existing report editing it then saving it with a new name For both methods the procedure for designing a report and customising its content is the same Adding elements to the report This section shows how to add elements to a report select any of these elements and move or align them on the screen Add all the required elements to the report then move them to the correct positions before going on to the next stage The Toolbox is used to design reports E General Text Picture Parameter Calculation Frame HOPS Particle Image Grid Graphs Distribution Graph Trend Graph Classification Graph EEE Scattergram Tables Classification Table The Toolbox can be moved anywhere on the screen It can be switched on and off using the View Toolbox command on the Report Designer menu Page 13 4 MAM 0410 Creating custom reports
130. logical parameter across a selected set of records For best results the measurements should W be produced by the same SOP For example if one record s SOP used a 100 pixel trash size and another record s SOP 200 pixels it is not ideal to compare the two records Other parameters which should be the same are scan area size objectives used intensity level and image processing settings B have similar numbers of particles B use the same filters The tab shows record similarity means undersize and frequency curves Morphologi Example 2 vmes E File Edit View Measure Tools Security Window Help iD m i Classification Particles Dm E ie B Records ae Scattergram A 37 Comparison acy Classification chart IM E Classification table IV acs Particles IV Group Record Sample Name SOP Name Date Edited Particles Class Count Glass Spheres Glass Spheres Class Count Salt Cryst Salt Glass Beads 2 09 November 2006 10 06 04 04 2 20 0 8 3 Salt minus Glass Beads 09 November 2006 10 06 04 a2 0 0 0 00 Salt minus oblates 09 November 2006 10 06 04 Tue S a gt i sa lt Selection Clusters Select All Select Grouped Auto 2 v Parameter Variability Dendrogram Trend Plot RUNE Cluster by Variability O Area un CE Diameter um Length um R3 Salt Glass Beads 2 Max Distance pm Perimeter p m SE Volume prr O Width um oil LM As
131. logue This offers a quick way of moving through the SOP Editor just click on any branch in the tree to move to that stage Once one or more objectives are SOP Editor New SOP selected the Optics selection area Fle Edit Help expands as shown below QD ala v Sample details Pre measurement settings Measurement central Sample carrier Illumination Optics selection B 20 1 8um 100m ul Sample Dispersion Unit Threshold Illumination ipic Optics selection Analysis settings Analysis settings Filters Filters Classification Classification Post measurement settings Reports Post measurement settings Reports The other buttons on the SOP Editor toolbar are Button Function gt Starts a new SOP m Opens an existing SOP L4 Saves the current SOP Page 6 6 MAM 0410 Measurement tutorial Chapter 6 Button Function l Saves a modified SOP with a new name The following buttons are only available in the Filters and Classification dialogues Copy copies a selected filter or class to the Windows clipboard Paste copies a selected filter or class from the Windows clipboard k Undo undoes the last action gt Delete deletes the selected filter or class Each SOP Editor dialogue has a Help button advising how to complete it Some SOP Editor dialogues have tooltip help on their parameters To view this move the cursor over the Y symbol in the dialogue To create a new S
132. looking for the smallest diameter particle It may be necessary to change to a higher magnification to estimate the diameter accu rately Make a note of the approximate diameter Ensuring the quality of the dispersion The general objective is to achieve a dispersion with as many particles as possible in each frame without them touching each other Although the software algorithms can split touching particles it is best to ensure that particles do not physically touch Note To capture the frame image to a graphics file use Image Save As In the Save dialogue select the Scale bar option if using the image in a report etc but not if the image file is to be analysed again the scale bar would appear as several particles When all is complete click the OK button to close the Microscope Manager Morphologi G3 Page 6 5 Chapter 6 Measurement tutorial 2 Creating an SOP Having chosen the correct magnification and checked the dispersion quality the next step is to create an SOP to make the measurement This means using the SOP Editor as described here Each SOP Editor dialogue has a Help button advising how to complete it Note After creating the first SOPs in this way try creating a new SOP by editing an existing one This is described at the end of this section The SOP Editor comprises several dialogues the user can step through using the j and Q3 buttons The tree structure shown below appears in the left hand side of each dia
133. low on the left then place the sample cartridge in the top of the sample chamber like this ill 8130 5 Screw the screw on cap by hand one turn until it s tight forming an air tight seal ill 8131 Page 4 4 MAM 0410 Preparing the sample Chapter 4 6 Push the chamber into the clamp on the G3S as shown below ill 8125 7 Push the air pipe O into the top of the chamber If it does not fit check that the slider is pushed in correctly EE Ep wa 8 The sample is ready for dispersion ill 8126 Morphologi G3 Page 4 5 Chapter 4 Preparing the sample To load a sample for dispersion reusable sample cartridge Preparing the reusable sample cartridge follows a similar procedure to above except the sample is loaded into the cartridge before the top foil disc is inserted 1 If the sample cartridge has been used before remove any traces of the old foil from the previous dispersion Insert and secure a foil disc to the base of the sample cartridge Load the sample in to the sample cartridge as described in the previous section Q Insert and secure a foil disc to the top of the sample cartridge Place the sample cartridge in to the sample chamber as described in the previ ous section Continue the procedure from in the previous section Making the dispersion Sample dispersion with the SDU is only possible using the software There are two ways to work
134. ment 5 24 Joystick 5 25 Measure menu 5 11 Security menu 5 12 Tools menu 5 12 View menu 5 11 Window menu 5 13 Menu commands in text 1 4 Menus in Microscope Manager 5 24 main 5 8 Method development 6 3 Pace iii Index Microns per pixel 7 5 Microscope focussing 6 32 initialisation 6 2 Microscope Manager controls 5 21 described 5 20 menus 5 24 modes 5 20 SOP mode 5 23 using 6 3 Model number 1 5 Monitors 3 2 3 15 Morphologi G2 components 3 3 functions 2 1 Morphological parameters defined 2 7 Multiview mode 5 20 N National Physical Laboratory 3 11 Navigation toolbar 5 14 Number basis 2 14 Number based resolution 2 14 Number based statistics 2 1 O Objective changing 6 5 in nosepiece 3 4 selecting 6 13 specification A 2 Objective nosepiece 3 4 Operator tasks 1 3 Optic button 6 4 Optical unit 3 4 Optics selection dialogue 6 13 Optional plates D 1 Options dialogue 6 14 Orientation 2 8 Overlap See Frame overlap P Packaging B 5 Page Layout reports 13 2 Parameter definitions 2 7 Page iv Morphologi G3 in report 13 8 Parameter Variability pane 8 15 8 16 Parameters tab 9 5 Particle agelomerations 2 16 context view 8 12 exporting 9 6 needle shaped 2 17 segmentation 6 18 sorting 8 7 Particle filtering 8 8 Particle image grid in report 13 11 properties 13 11 Particle in Image dialogue 8 7 Particle in image dialogue 8 11 Particle shape overview 2 3 Particle size defined 2 2
135. ment from files E Measurement from file X Close Summary Particles 597 CE Diameter um Mean 8 22 CE Diameter um Std Dev 3 13 10 Percentile 5 25 50 Percentile 7 23 90 Percentile 12 33 10 100 CE Diameter um E Diameter um v welume Transformation Classifications lt Impurity gt lt Acceptable gt Unclassified 200 300 Number of Particles S Multiview Result Video 11 When the measurement is complete the Status bar will show this and report the number of particles found The smaller windows within the Measure ment Manager give details of the measurement as described in Chapter 5 Click the Close button 12 The data record will appear in the Record tab The record can be viewed and edited in the same way as other records Page 7 12 MAM 0410 Viewing the results Introduction When a measurement finishes the results can be reviewed This chapter describes the various ways of displaying the final measurement result It shows how results are displayed in the Records Scattergram Comparison and Report tabs and how to use these to understand completely what the result shows The sections are Morphologi G3 The Records tab explains how to select move and copy records and how to batch print that is to print reports for multiple records The Scattergram tab describes this tab and shows how to use it to select particles create filters and d
136. menu commands will be greyed out Page 5 8 MAM 0410 Software features Chapter 5 File menu The File menu has the following options Command New gt gt Measurement gt SOP Function This has the following sub options Creates a measurement file where the measurement records results will be stored Starts the SOP Editor for creating an SOP Open gt gt Measurement This has the following sub options Opens an existing measurement file As a shortcut a list of the most recently used measurement files is shown at the bottom of the menu Click any of these to open it gt SOP Opens an existing SOP for editing Close Closes the current measurement file Save as Saves the existing setup as a measurement file or saves an opened measurement file with a different file name Export Result Exports information on measurement records from a measurement file to a software package such as Excel or Wordpad see Chapter 9 for details Export Particles Exports information on each particle in a measurement file record for further processing in a software package such as Excel or Wordpad see Chapter 9 for details Export Images Produces a bitmap file for each particle in the sample Print Setup Configures the printer and paper setup Print Preview Shows how a report will appear when printed Print Prints a report Morphologi G3 Batch pri
137. n moves the microscope to focus on this particle on the slide if the slide is still there e Moves the target in and out Indicates whether Bright Field B or Dark field D illumination is in use Controls The controls in the window are Optic Optic button the text on this button changes to show the current objective Use it to select the objective to use from a list The motorised nosepiece rotates automatically when an objective is selected Once the objectives have been characterised when the user changes between objectives the system automatically compensates for small variations in focus that 1s it corrects for parfocality errors Light WB Darker Brighter buttons to decrease or increase the light intensity in 5 increments click Darker or Brighter respectively B Custom light calibration To pre select a custom value enter the required figure and then click the Custom light calibration button The system now performs a light calibration see Chapter 6 B Standard light calibration button resets the light intensity It may be nec essary to do this after a change of objective see Chapter 6 After the intensity is reset the button displays a v Morphologi G3 Page 5 el Chapter 5 Software features X Y Position Z Position Move to preset position button use this to move directly to predefined X Y positions a grating the slide plate centre or one of its corners the light calib
138. n of the parameter illustrating what a Statistical Process Control SPC plot would look like Use it to determine SPC information In this example a cut off at 0 1 would separate the two groups of particles Frequency curves The morphological parameter that shows the big gest difference value between two records is by default selected for showing the two distribution plots The user can select another parameter The curve for each cluster is shown in a different colour in accordance with the dendrogram Undersize curves This is also known as a cumulative oversize curve or result under plot It displays the per centage of the particles which are under a certain value The curves for members of each cluster are shown in the same colour Trend Plat 1 1 l I T I l I I 1 1 D T 0 05 0 1 0 15 0 2 Elongation Frequency Curves Elongation Undersize Curves 0 0 2 1 4 Elongation Page 8 17 Chapter 8 Heport tabs Viewing the results To display measurement information for one or more records in report form select these then select the relevant Report tab The choice of graph type depends on user preferences and the specific objectives of the analysis For example a cumulative graph type may be more appropriate for comparing multiple results The available Malvern default reports are listed below These are all number based e
139. nd buttons High contrast darker particles normally need a lower threshold lighter crystal line samples generally need a higher threshold The graded scale under the slide bar is a reminder of this The default setting picks up the darker parts of the image Note Left click and hold the mouse button down on the large image to view the original non thresholded image This helps decide the threshold to use 5 If analysing more than one file select each image in turn from the Image file list to check the threshold looks OK After this click the button 6 The Analysis settings dialogue is displayed Image File Analysis Qs Blo X Image Files Sample details Threshold Analysis ID Filters 2 00 Classification Analwsis J 299 A Trash size Minimum number of pixels 10 v Segmentation method Watershed Fill holes Enable hole filling Size bands Morphologi G3 Page 7 7 Chapter 7 Page 7 8 Measurement from Files Specify a value for any of the following parameters Analysis ID the default should be the highest number in the list Only select a lower number for backwards compatibility with data from older versions of the software For example 2 00 gives compatibility with data from Morphologi G3 releases prior to 6 0 and 1 00 gives compatibility with the PVS 830 a predecessor of the Morphologi G3 Trash size discard any particles smaller than this size prxels Use this
140. nd the oil are similar it can be difficult to get good contrast This prevents the par ticles from being detected well In this case try oils with different refractive indexes MAM 0410 Preparing the sample Chapter 4 Oils typically used for this type of dispersion are mineral oil silicone oil e g poly dimethylsiloxane paraffin oil and Iodo napthalene oil Note When using Iodo napthalene oil always use glass pipettes and vials Water dispersion If samples are not soluble in water it is also possible to adopt a similar technique using water as the dispersion media often with a small amount of surfactant added to prevent agglomeration of particles In this case it may also be necessary to put oil around the edge of the cover slip to prevent the water evaporating during the meas urement Morphologi G3 Page 4 11 Chapter 4 Preparing the sample Page 4 le MAM 0410 Software features Introduction The standard Morphologi software comprises two modules Morphologi G3 The main Morphologi G3 application described in this chapter This controls the system during a measurement and then processes the measurement data It displays the results and allows reports to be printed The Report Designer for creating custom reports to display the results This is detailed in Chapter 13 This chapter introduces the features of the main software application It describes The main window describes the appearance of
141. nd the sample with the joystick Adjust the Z axis position to focus using the Z position control For fine focus adjustments use the joystick s focus knob For larger Z changes adjust the Step Size slide bar moving it downwards decreases the step size and then click the up down buttons Each click on these buttons moves the objective up or down by the Step Size As the focus improves reduce the Step Size or switch to the joystick Note As requirements differ depending on the sample the following are just guidelines Look at several frames This part of the method development has two aims as detailed below W to establish the correct magnification to use for the sample B to check the dispersion quality Note Q Z Stacking is an option that can be used to provide a sharp image where the objective s limited depth of field is an issue See Chapter 5 for detailed information MAM 0410 Measurement tutorial Chapter 6 Choosing the correct magnification Choose a medium magnification for example 10x then Note It may be necessary to modify the focus Z position slightly whenever the objective is changed m Move around several frames looking for the largest particle available If a particle larger than 40 of the field of view is found change to a lower magni fication Make a note of the approximate diameter of the largest particle using the scale bar to estimate this W Move around several frames
142. ng corrects for this Select the command Instrument White balance At this point the image may appear black and white but this is normal it is not a problem 4 Use the standard Microscope Manager controls or the joystick to move to and focus on the relevant part of the slide 5 Morphologi G2 users only If the image colour does not appear correct now use the command Instrument Manual lamp control to enable manual use Page 8 27 Chapter 8 Page 8 28 Viewing the results of the Lamp dial on the microscope Turn the lamp intensity down and repeat steps 3 and 4 It may be necessary to adjust this several times until a good result is obtained 6 Select the command Image Save as and specify a file name After capturing images After capturing the required image s select Instrument Colour mode again to return to normal monochrome mode That is this command toggles between monochrome mode and colour mode The Microscope Manager always opens in monochrome mode MAM 0410 Exporting results Introduction This chapter describes how to export Morphologi data for use in other applications It covers m Direct copying of data Exporting result data how to export one or more records to a file Exporting particles how to export information on each particle in a record Exporting images how to produce an image of each particle in a record Creating export templates these control the way data is exp
143. ng particles with the scattergram 1 Page 8 4 In the Scattergram pane shown below use the major filter 9 to select the parameter to filter on Area Pixels ji CE Diameter pm 100 1000 10000 100004 1 Circularity ill 8069 MAM 0410 Viewing the results Chapter 8 2 Using the parameter selectors adjacent to the two graphs select the two parameters to plot 3 The scattergram plots the position of each particle based on the two parameters selected 4 When a particle in the Particles pane at the right hand side of the tab is selected its position in the scattergram is highlighted as shown below Click on a few particles there and look at the scattergram In this example particle Id 8 is selected in the Particles pane on the right The pointer shows its location in the scattergram 5 Use the mouse to drag a rectangle around the region of most interest Particles in the selected area become red Only particles in the area selected are shown in the Particles pane at the right hand side of the tab In the example below dragging two different rectangles shows that particles in the area near the origin are circular but in the area near the bottom of the scattergram they are irregu larly shaped Area selected Particles displayed E pm M 5 1 2 Sg 7 S 1156 To close the selection rectangle right click within the scattergram pane and select Clea
144. ng the product against and being found to be compliant with FCC CER 47 Part 15 March 2003 Class A digital device The device complies with part 15 of the FCC Rules Operation is subject to the fol lowing two conditions 1 This device may not cause harmful interference and 2 this device must accept any interference received including interference that may cause undesired operation Q Page C e Note This equipment has been tested and found to comply with the limits for a Class A digital device pursuant to part 15 of the FCC rules These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment This equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause harmful interference in which case the user will be required to correct the interference at his own expense Note Changes or modifications not expressly approved by Malvern Instruments Limited could void the user s authority to operate the equipment MAM 0410 Regulatory statements Appendix C Canadian Regulatory Information This applies to Canada Only This digital apparatus does not exceed the Class A limits for radio noise emissions from digital apparatus set out in the R
145. ngs Font Upper Limits Lower Limita Trend Plats click on a cell to modify Farameter Axis Multiplier CE Diameter Mean urn 1 Axis Against horizontal axis Parameter Length Maximum pm L Use the Add button to select one or more parameters for the Y axes then specify which parameter to plot these against Chapter 13 and the online help give details For a trend graph the Axis Settings and Fonts tabs are as described above for a dis tribution graph Page 8 23 Chapter 8 Viewing the results Use the Options tab to specify B The key position m Whether to use graph tips m Whether to display points relative to the time of the first measurement Upper Limits Lower Limits tabs This controls the display of warning or action limits expressed in percentage terms and shown as horizontal bands across the graph Properties Properties Options Avis settings Font Upper Limits Lower Limits Limits will be displayed against the Y1 axis Show Warning Limit Upper Warning Limit Shaw Action Limit Upper Action Limit Cancel Help The following example shows how limits are displayed on a report This shows both warning and action values set for upper and lower limits Trend Graph 40 CE Diameter Mean tum 11 12 13 14 15 16 17 Record Page 8 24 MAM 0410 Viewing the results Chapter 8 Interpreting classes in the display The Classification Chart report
146. not contribute to the analysis for example particles under 100 pixels in area do not help in an analysis of shape so should be excluded if shape is the primary factor In this example the particles could be filtered out using the Area pixels parameter Filters to exclude particles can be defined during the SOP design stage or created and applied after a measurement The Scattergram provides an easy graphical tool for determining filter settings Filters use OR logic so a particle is excluded if it meets any of the filter criteria Classes In Quality Control applications the number of impurities in a sample may be important These may include particles of dust fibres and bubbles For cosmetics the number of large angular particles may be important Any of these groups which has consistent properties can be defined as a class The number of particles in each class can be reported during the analysis A class can be defined then excluded that 1s used to filter fines out For example a class bubbles can be defined by a combination of the following two properties B HS Circularity value of over 0 95 WB Intensity SD Standard Deviation value of over 0 5 Classification uses AND logic meaning that a particle is only a member of the class if it meets all of the filter criteria Particles can be a member of more than one class The Scattergram provides an easy graphical tool for defining classes Morphologi G3 Page e e3 Cha
147. ns ffnen und die oberen Schaumeinlagen entfernen Abrir los costados de la caja y extraer las piezas intercaladas de espuma de la parte superior Abra as abas da caixa e retire o enchimento de espuma que est em cima MAM 0410 Unpacking instructions Appendix B 5 Unpack the components as shown in the illustration below D baller les l ments de la facon d crite dans l illustration ci dessous Die Komponenten nach unterer Abbildung entpacken Desempaquete los componentes siguiendo la ilustraci n Desempacote os componentes como mostrado na figura abaixo D G3 Instrument main body 2 XY stage Camera and tube Cables 9 Consumables Installation kit Lamp house x2 2 Joystick Z Drive assembly 9 Chamber PSU P Top cover 2 Camera cover 9 Rear cover Base cover ill 8216 Morphologi G3 Page B 3 Appendix B Unpacking instructions Lift the instrument out of the carton and place on the workbench If handles are fitted later versions only use these to lift the instrument Soulevez l instrument pour le sortir du carton et posez le sur le banc Si des poign es sont install es derni res versions seulement utiliser les pour soulever l instrument Das Ger t aus dem Karton heben und auf der Werkbank aufstellen Wenn das Gerat mit Griffen ausgestattet ist nur sp tere Modelle sollten diese zum Anheben verwendet werden Levantar el aparato sacarlo de la caja y col
148. nt Prints reports for a number of measurement records simul taneously Create PDF Creates a pdf file from a report This is only enabled if the 21 CFR part 11 feature key has been installed Exit Shuts down the software Note Q Right clicking anywhere in a Records tab displays available commands These are from several different menus Page 5 9 Chapter 5 Page 5 10 Edit menu Software features For Comparison tab components use Edit Copy Dendrogram Edit Copy Trend plot etc Use this menu to manipulate records in a Records tab and to copy graphs Command Function Cut Copy Paste These are the standard Windows options Records can be and Delete moved to from the current file or other measurement files Copy Scattergram Copy catiergiam Copies these Scattergram tab components to the and Histograms i Windows clipboard Copy Particle window Copy Parameter Variability Panel Copy Dendrogram Copy Trend Plot Copies these Comparison tab components to the Copy Frequency Windows clipboard Curves Copy Undersize Curves Undo Redo Undo repeat the last command Copy lt type gt In a Report tab copies a complete graph of the specified Graph type into the Windows clipboard for pasting into another application for example Microsoft Word EON Edits sample name comments for a record Documentation Extract SOP Views the settings for the selected measurement record The SOP Editor d
149. number This also gives the exact values for any data point on the report Zooming in out To zoom into an area of a graph in a report hold down the left mouse button and move the mouse to draw a marquee around the area to be enlarged To zoom back out simply click the left mouse button anywhere on the graph Screen layout and printing reports To accommodate the different aspect ratios of the printed page and the computer screen for each report two views are created as shown below m shows the printed version portrait E shows the computer screen version landscape ill 7448 Use File Print preview to see what the printed page will look like The screen and print formats can be configured separately to report different information A report file has the extension vrep and holds both the screen and print versions These files are stored in the Report Pages folder MAM 0410 Viewing the results Chapter 8 Modifying graph formats Morphologi G3 To modify temporarily until the software is closed down a graph in a report right click on it to display its Properties dialogue To modify the report perma nently use the Report Designer to create a new report type Its tabs are described briefly below and the online help gives full details Distribution graphs The tabs on the Properties dialogue for this type of graph are described below Distribution Graph Properties Font Upper
150. o cover the scan area ABCDE Fe lt _ gt ill 7447 Again the tiered rectangles underneath show which particles are measured by each frame This time all the particles are completely contained within at least one frame Provided that all particles in the sample are smaller than the overlap field of view all the particles in the sample will be detected In this illustration particle in frame B would not be counted as it has already been measured in frame A The particle in frame E is the largest that can be meas ured as this is almost as large as the overlap itself indicated by Page e enoO MAM 0410 What is the Morphologi G3 Chapter 2e Morphologi G3 Particle stitching Particle stitching provides an alternative method to Overlap for capturing more particles that cross image borders Like Overlap it effectively extends the upper size measurement range of the objective ensuring that larger particles by nature those that cross borders more frequently are counted Particle stitching separately records all items that cross boundaries and then uses shape recognition algorithms to match them together into single particles Unlike simply tiling entire scan regions together this method removes unwanted back ground before stitching improving analysis speed See illustrations below Before particle stitching particle straddles frames 4 Disconnected
151. o filter out the particles in the darker area that this draws In the following example the user has dragged the right hand margin to the left to filter out a tail of large particles Area Pixels w Filterz 100 1 000 10 000 100 0 4 Release the mouse button and look at the Filters tab This shows the exact numeric value of the filter created like the following MAM 0410 Viewing the results Chapter 8 Selection Classification Filters Parameter Operator Value 1 Value 2 Filtered aut Area Pinels Aro 143 The Filtered out column shows how many particles the filter excludes If it s necessary to change the filter drag again or type a value into the Value n box on the Filters tab To add a second filter select a new parameter and repeat the process A red cross watermark marks the particles excluded by filters in the Particles pane Clicking the button removes these particles from view in the dis play Filters can be copied between SOPs or measurement files using the Edit Copy and Edit Paste commands Filters can also be set up using just the Filters tab but this takes longer Use the same process described for the SOP Editor s Filters dialogue in Chapter 6 To produce a new record with the filtered particles excluded 1 3 Morphologi G3 Click the gh button The Documentation dialogue asks for a description of the new record include details of any filter settings us
152. o focus the microscope When multiple objectives are selected this step is repeated for each objective Focus on sample amp Y Position Control Position Control Light source Step Size Move Frame MT EPI DIA Advanced Z position tmr E dv Move Target Goto Zero Position Bright Field 0 029 If using the dialogue the Z Position Control panel controls the vertical movement of the objective First set the size of increments by dragging the Step Size slider The value of the step size is shown below the slider Next click on the up down buttons or the joystick to move the objective Each click moves the objective by the step size specified The value below the buttons shows the current position in mm from the Z origin Use the X Y Position control buttons to find a frame containing one or more appropriate particles If two or more objectives are being merged focus for each in turn starting with the lowest magnification objective For low mag nifications or when using just one objective focus on big particles For high magnifications focus on small particles Fit to window check box this option selected by default allows the user to navigate the preview image smoothly in order to identify a region of interest on the sample Once the region of interest is centred in the preview de select Fit to window This provides a closer view of the sample making it easier to obtain a more precise level of focus Note Q If
153. ocarlo sobre el banco de trabajo Use las asas solo presentes en las versiones recientes para mover el instrumento Tire o instrumento da caixa e coloque o na bancada Caso haja al as somente vers es mais recentes use as para levantar o instrumento Page B 4 Never try to lift the instrument without the help of others Ne tentez jamais de soulever l instrument tout e seul e Bitte versuchen Sie niemals das Ger t ohne Hilfe einer zweiten Person zu heben Stets die korrekte Hubart verwenden um eine R ckenverletzung zu vermeiden No se debe intentar nunca levantar el aparato sin ayuda de otras personas Aplicar siempre las t cnicas de elevaci n adecuadas para evitar da os en la espalda N o tente nunca levantar o instrumento da caixa sem o aux lio de outros use sempre t cnicas de levantamento adequadas para evitar les es nas costas MAM 0410 Linpeckimng instructions Appendix B 8 Once assembled never lift the instrument by its covers or the focus knobs Une fois assembl ne jamais soulev l instrument par ses couvercles ou par les boutons de r glage Das zusammengebaute Gerat niemals an den Abdeckungen oder am Fokusknopf anheben Una vez ensamblado nunca use la cubierta o los mandos de enfoque para mover el instrumento Uma vez montado nunca levante o instrumento pelas tampas ou ajustador de foco 9 Always place the instrument on its feet Placing i
154. of the sample Joystick for moving the XY stage manually The following sections describe the main components Page 3 e MAM 0410 Hardware features Chapter 3 The instrument The Morphologi G3 G3S has the following major components ill 8120 These are Optical unit Back panel connections Integral SDU G3S only Precision XY stage Sample plate holder and sample plate FireWire digital camera under removable part of cover 40V power supply for control electronics OQoocoo2ogo Joystick for moving the XY stage manually Instrument identification label this gives the Morphologi G3 model and its serial number To access the label remove the triangular cover Please quote these numbers when contacting Malvern Instruments not those on the Nikon label on the back panel This section summarises the functions of these components and shows how to connect the instrument to the computer Morphologi G3 Page 3 3 Chapter 3 Page 3 4 Hardware features D Optical unit The main components of the optical unit are shown below A J ill 8121 The components are 0000009 Main optical body Objective nosepiece houses a set of objectives with different magnifications Automatic changeover between objectives is controlled by the software Back panel connectors Lamp houses the Essentials Manual shows how to change their bulbs Digital camera Integral SDU
155. ogether with details on how to configure SOPs for their use See Chapter 6 for more informa tion on running an SOP Summary of plates m Filter Plate for holding two filters primarily used in for foreign particle detection WB Wet Cell for measuring particles dispersed in a liquid B Two Slide Plate two slide plate for holding 75x50mm slides Page D 1 Appendix DO Optional sample plates Filter Plate The Filter Plate is primarily aimed at foreign particle detection analysis It provides a platform in which to clamp two separate samples prepared on filters and is availa ble for two filter diameters 25mm and 47mm Part no Description MOR5300 Filter Plate 25mm MOR5310 Filter Plate 47mm Overview The parts of the Filter Plate are shown below o b ill 8214 The features illustrated are Filter plate base Filter clamp O Filter Filter recess Thumb recess Page D e MAM 0410 Optional sernple plates Appendix LJ Morphologi G3 Tweezer recess Filter clamp apertures Filter protective cover not supplied see Recommended filters on page 5 Usage Sample insertion Ensure that the Filter Plate is clean before inserting a sample T Place the Filter Plate on a suitable surface and open one of the filter clamps using the thumb recess to lever the clamp open The magnetic closure may require moderate force to break the sea
156. or Font Calculation ll Calculation Editor Option Explicit Private Function Calc As Variant Enter your calculation here Calc No Calculation End Function The Properties dialogue is similar to that for a text element but with the addition of a General calculation tab that enables editing of the calculation Press the Edit button to open the Calculation Editor Page 13 9 Chapter 13 Page 13 10 Creating custom reports Calculation Editor Use the Calculation Editor to load an existing calculation for editing or to create a new calculation The language used Sax Basic is similar to Microsoft Visual Basic for Applications VBA Note This is a prototype version which is available for use now but will be fully supported in future releases Select the browse icon Sj to choose functions from the library lists which can be used to build up calculations All details about a measurement and how it was per formed can be accessed in the available data type lists i e size results measurement and result details More information is available from the Calculation Editor help buttons Button Function bs Describes the Sax Basic language j Describes how to use the Calculation editor Example This example shows the simple calculation used to print the optics used at the top of some Malvern reports Option Explicit Private Function Calci s Variant Enter your calculation here
157. or exporting 9 8 Tensioners 3 15 Text in reports 13 7 Threshold dialogue 6 16 7 6 Threshold settings E 1 Threshold slide bar 6 17 7 7 Tilt compensation in SOP design 6 33 Toolbars 5 14 Toolbox 13 4 Toolbox reports 13 4 Tools menu 5 12 Tools Microscope 5 20 rash size 6 21 rend graph 13 12 rend graphs 8 23 Trend Plot 8 15 Trend plot 8 17 Two slide plate D 10 U Undersize curves 8 17 Unpacking instructions B 1 Upper Limits graph 8 23 8 24 User Page vi Morphologi G3 adding 10 6 definition 10 6 logon 10 7 User group adding 10 4 V Video mode 5 20 View menu 5 11 Viewing 8 12 Viewing particles 8 6 Viewing results 8 1 View Workspace command 8 3 vmes file 5 2 Volume basis 2 14 Volume Weighted Mean 2 14 vsop file 6 26 W Website 1 6 Wet Cell D 6 White balance command 5 24 White balance command 8 27 Width parameter 2 9 Window Classification 5 19 Distribution 5 18 Live picture 5 19 summary 5 18 Window menu 5 13 Workspace Malvern default 8 2 overview 8 3 toolbar 5 14 X XY stage components 3 10 XY Joystick control 5 22 Z Z Position Control 5 22 6 4 6 32 Z Stacking defined 2 22 in Microscope Manager 5 22 in SOP 6 15 Zooming in report 8 20 MAM 0410
158. or number distributions Page 2 13 Chapter 2 Page e i4 What is the Morphologi G3 B D n 0 1 is the size of particle below which 10 of the sample lies B D n 0 9 is the size of particle below which 90 of the sample lies B D 4 3 is the Volume Weighted Mean or Mass Moment Mean Diameter B D 3 2 is the Surface Weighted Mean also known as the Surface Area Moment Mean Diameter or Sauter mean Number based resolution Some applications particularly those where the detection of small numbers of rela tively small particles fines or the detection of foreign particles 1s important require the resolution of a number based system Not all applications require this number based sensitivity if not it usually makes sense to benefit from the speed and convenience of ensemble methods which are volume based Volume basis Ensemble particle sizing methods usually provide data on what is known as a vol ume basis This means that the contribution each particle makes is proportional to its volume large particles dominate the distribution and sensitivity to small parti cles 1s reduced as their volume is so much smaller than the larger ones This graph shows a sample measurement displayed on a volume basis Volume transformation CE Diameter pm smoothed over 11 points CE Diameter um Record 1 sample 2 Number basis Image analysis provides data on what 1s known as a number basis This means that the contribution
159. ore running the measurement setting a higher value in the Intensity tolerance field will speed up the calibration process Click the button when complete MAM 0410 Measurement tutorial Chapter 6 9 The Optics selection dialogue is displayed SOP Editor New SOP File Edit Help peack 1i A Sample details Pre measurement settings Measurement control Select the optics based on your particle size range Sample carrier Sample Dispersion Unit Illumination Optics selection 50x 0 5um 40m Analysis settings Filters Classification Post measurement settings Reports 20x 1 8m 1001m 10x 3 5um 2101m 5x 6 5um 420m 2 5x 13pm 1000um Select one or more sets of objectives which are suitable for the range of particle sizes in the sample based on the approximate particle size range established in Choosing the correct magnification earlier With broad particle size distri butions it may be necessary to merge the data from several objectives The numbers in brackets after each objective show the suggested size range When an objective is selected its band in the graphic changes colour If a sec ond objective is selected the shading of the two changes as shown below The blue area in the window shaded the darkest grey below shows which objec tive will be used for any particle size In this example the 5X objective will be used for sizes between 6 5um and 430um and the 10X only for particl
160. orised changes the software can be configured to limit each user s access to functions like modifying SOPs Users are assigned operating per missions that allow or restrict access this is fully explained in Chapter 10 The menu options are as follows Command Logout Function Logs out the logged on user Configure Security Sets up user accounts permissions and groups Change Password Changes the user s password Audit Trail Displays the current audit trail only if 21 CFR part 11 is in use MAM 0410 Software features Chapter 5 Window menu Use this menu to alter the view characteristics of any open measurement file win dows Command Function New window Opens a new view of the current measurement file Cascade Overlaps all open windows Tile Fits all open windows into the available space Arrange icons Lines up all open icons Windows Opens a list of all current windows and provides more commands for reorganising their display Help menu The Help menu options are as follows Command Function Help Topics Opens the help file Tutorials Offers a selection of tutorials Gives hints on how to use the Morphologi software a dif ferent tip appears each time this is selected The user can turn off the Tip of the day dialogue that appears when the software starts Tip of the day Shows the software version installed Quote this when About Morphologi contacting Mal
161. ormation check box to show volume based rather than number based measurements Summary window The Summary window shows the number of particles detected and the mean standard deviation and main percentiles for the parameter selected in the list under the Distribution window Summary Particles 12 CE Diameter um Mean 306 78 CE Diameter im Std Dev 49 38 10 Percentile 240 22 1 50 Percentile 313 56 90 Percentile 353 47 Y MAM 0410 Software features Chapter 5 Classification window The Classification window shows the number of particles in each class if classes have been defined Classification lt Impurity gt lt Acceptable gt Unclassified i FOO 400 600 800 1000 1700 Number of Particles In this example two classes were defined Impurity and Acceptable Particles which are not in any of the classes appear as Unclassified Live picture window The Live picture window shows the image on the plate while a measurement is in progress as shown on the second monitor It shows the same picture as the video window shown below Afterwards it will be blank ae Measurement 4 g 9 x Documentation Close Morphologi G3 Page 5 19 Chapter 5 Software Features Mode tabs Three modes are available B Multiview the window is split into four as shown at the start of this section This is the default B Result the Distribution and Summary windows
162. orted Morphologi G3 Page 9 1 Chapter 9 Exporting results Overview The values of any of the parameters from one or more records as well as graph data from a report can be exported for use in other applications such as Microsoft Excel Word and Wordpad There are three types of File Export command exporting parameters particles and images respectively WB Fora complete measurement file or a selection of records File Export Result creates a text file which has user defined parameters as columns There is a row for each record Columns can correspond either to those seen in the Records tab or to user defined columns In the latter case a template is used to specify which record parameters are exported The Tools Settings Data Export Templates command allows the user to create and modify their own templates B Fora single record File Export Particles creates a text file with a row for each particle and a column for each size shape parameter WB Fora single record File Export Images creates a bmp file for each particle Note that this can create a large number of large files if there is a large number of particles in the record The default location for these exported files is C Documents and Settings All Users Shared Documents Malvern Instruments Morphologi G3 Export data Information can also be copied directly using drag and drop or the Windows clip board B One or more records can be selected in the Reco
163. orts m Exporting results for use elsewhere Part 2 Supervisor s guide The Supervisor s guide concentrates on administration and extension of the Mor phologi G3 functionality Topics covered are m Security setting up administrators users and user groups m Managing SOPs extracting modifying and distributing SOPs making them available to other sites B Organising the workspaces which control what is displayed on the screen W Creating custom reports these can include parameters which do not appear in standard reports and also incorporate company information etc The supervisor should also read the Operator s guide Page 1 2 MAM 0410 Introduction to this manual Chapter 1 Part 3 Appendices Appendix A gives the specification Appendices B and C contain supplementary information not necessary for the general operation of the system Appendix D provides information on the optional sample plates Warning IN The instrument and the samples to be measured may be hazardous if mis used Users must read the Health and Safety information in the Essen tials Manual before operating the system Access to the instrument Morphologi G3 This manual refers to the various people who will have access to the instrument as follows Malvern Instruments personnel Malvern Instruments personnel service engineers representatives etc have full access to the instrument and are the only people authorised to perfo
164. out particles later using filters B Segmentation method watershed segmentation is enabled by default This works well for spherical particles but not for example for crossed needle like particles Further segmentation methods suitable for different sample types will be developed in future W Fill holes select the check box to include lighter areas within shapes When hole filling is turned on the middle of a doughnut shaped particle is Morphologi G3 Page 6 el Chapter 6 Measurement tutorial counted as part of the shape When hole filling is on a doughnut shaped particle will have a high area without hole filling it will have a low area m Size bands for speed the graph displays bins or bands rather than the raw data on each particle The default defines 1000 bands covering the full range of the instrument This can be modified if more graph resolution is required in a specific range If required click the Advanced button to change the graph size bands using this Size bands dialogue the online help gives full details Size bands Lower size um Upper size um Bands T A A A xi 2000 1000 zi Spacing Size bands um When all Analysis settings are set up click the button Page 6 ee MAM 0410 Measurement tutorial Morphologi G3 Chapter 6 16 The Filters dialogue is displayed SOP Editor New SOP File Edit Help Qe OO elb Al Blox Sample details Pre measurement settings
165. p icon to start the software Note If the icon is not on the desktop open the Windows Start menu and select Programs Malvern Instruments Morphologi to start the software 1 If security is configured when the logon screen shown below is displayed type the user name and password if this is requested and click OK Chapter 10 shows supervisors how to administer security and set up users and groups User name 2d Type your user name to log on M a V e r n User name scoombe 2 The software opens If this is the first use of the software the Example Results vmes file will open If a measurement file was open in the previous session this will automatically reopen 3 The microscope performs an initialisation routine to zero itself This takes a few seconds during which some menu options are unavailable The instru ment connection icon on the status bar shows a clock during this period Ti Page 6 e MAM 0410 Measurement tutorial Chapter 6 1 Method development Method development means preparing the sample and configuring all hardware and software variables so that the sample measurement is reproducibly consistent Preparing the sample Prepare the sample as described in Chapter 4 Checking the sample dispersion Follow this procedure for all samples however they were dispersed once the sam ple plate has been placed in the stage To inspect the sample 1 Check that the microscope symbol in the Stat
166. particle sections are identified isolated and matched E Particle stitching is considerably quicker than using Overlap as much less scanning with the microscope is required to cover the same surface area However there are limitations in pattern recognition technology and this method is better suited to the analysis of elongated rather than circular particles It is advisable to experiment with both Overlap and Particle stitching on a small representative scan area initially to ascertain which method is more suited to the sample See Chapter 6 for details on how to enable particle stitching Note Q Unavailable on magnifications greater than 10X due to accuracy limits Due to the finite PC memory available for this process there is an upper limit to the length of particle that can be stitched This is dependant on magnification and complexity of particle shape As a guideline using the 5X objective a particle with a maximum distance of 10mm will be success fully stitched Page e el Chapter 2 Page e ee What is the Morphologi G3 Z Stacking Since the instrument s objective lenses exhibit a narrow depth of field due to their high magnification accurate focus cannot always be achieved across the entire body of a three dimensional particle Z Stacking is a utility that takes several images of the sample each at different Z heights and then overlays them to form a single composite image See illustration below Par
167. pears like this when added to a list CE Diameter Din x um x 7 Tosetthe argument value double click on the Argument Value box and type the value in Next alter the Column title to reflect the argument value by double clicking on it For the above example the user might enter 0 1 as the Argument value then change the Column title as shown below CE Diameter Din 10 um x 0 1 8 To change the order in which the parameters are displayed select a parameter and use the and buttons 9 Press OK to exit and return to the Configure export templates dialogue Press OK again there to save the template There are examples of three parameters with arguments in the above example of the Export Data dialogue MAM 0410 Pert e Supervisor s Guide Q Security Introduction Morphologi G3 Security principles describes the fundamental security principles Setting up the Administrator This is the person who will control security This chapter describes a a initially WB Security guidelines B Setting up user groups m Setting up individual users Page 10 1 Chapter 10 Security Security principles As the Morphologi G3 may be used by users of varying skill levels user access can be limited so that certain functions such as editing SOPs deleting records and edit ing results are reserved for specific users only Administrators users and permissions One or more persons are set up as
168. pect Ratio 05 06 07 08 09 1 Cireularity Aspect Ratio O Convexity R4 Salt minus Glass Beads 2 Elongation OHS Circularity O Solidity O Intensity SD Undersize Curves R5 Salt minus oblates 02 0 4 0 6 0 8 1 0 8 0 85 1 Record Differences Aspect Ratio Aspect Ratio 06 08 Aspect Ratio Dfenton To use the tab 1 Select the desired records in the Records tab and click on the Comparison tab The selected records are transferred to the tab s record pane If unsuitable records are selected for example just one record or any empty records a warn ing appears on the tab 2 Setup record groups if desired Records which are believed to have similar Page 8 14 characteristics can be grouped together To do this right click in the Group MAM 0410 Viewing the results Chapter 8 Morphologi G3 column and specify the group number to place the record s in In the example below two groups have been set up Group Recor Sample Name SOP Name Date 1 Salt Glass Beads 09 November 2006 10 06 04 1 3 Salt Glass Beads 2 5 2 54 09 November 2006 10 06 04 1 4 Salt minus Glass Beads 5 2 5 09 November 2006 10 06 04 2 5 Salt minus oblates 5 2 54 09 November 2006 10 06 04 2 b Salt minus oblates 5 2 54 09 November 2006 10 06 04 Click Select All to compare all the selected records or Select Grouped to compare only those records placed in a group in step 2 The software automatically compares all the morphologi
169. play llle 8 25 Capturing Colour images a 4 3 97 CARROS Mire a CST e cC 9 27 Pace ii MAM 0410 Morphologi G3 Table of Contents Exporting results EPO GUI OID se coi 9c eg di 0 dE aa RO e eoo GES a a A C dH EG Go 9 1 SIG WY Eo suu as ee E eta a dc dra Erde a ee oa d n 9 2 Direct copying of dala sus dox EX XR as AN a ae UR Re e eni 9 3 EXDOMING teslis uad uber qoc os eee EOS U ee ee acidic e E ds 9 4 Exporting particles 2455 v sura ador pee mte a Ro ed dg a ep ate ws Siva eto 9 6 Exbortind IMAGES e624 eset us ar Ree ox aH dac dab oec ea eed 9 7 Creating export templates sse rn 9 8 Part 2 Supervisor s Guide Security I EEOQU C HOFLIS 89 60 0097 dott c a C RERO CR CC RE RC ato e CAR 10 1 SecUntv DUMNGCID CS s cipe iix o BO RCRUM Boedo D Dea d 10 2 Setting up the Administrator be gr Ee acea CY eR RC EA E 10 3 Secunby guidelines s a a aede di ojo Edo SA ARE ee des 10 4 Setting up Usergroups lee hh hr 10 4 Setting up individualusers es 10 6 SOP management Iltro duction ac Eu ex e Re ae ed SCC RSS ERU SEO eU d 11 1 Extractng am OOP uuu tutrid a Sor sd PR YE UR V CRUS d Ec be iod 11 1 Moditying an SOP iss eti e ead E bata xcd bs adeb a ed 11 2 Distributing anm SOP Sav user bar cet dE cade dtc seed sod Sore dew 11 2 Workspace management IF EEOCIUIC EI OTI E dca de o wi de pot ct toa Ha dit Mace ge d c So ede de e eet a e 12 1 How to use workspaces eee hh han
170. propriate concentration of sample in the oil Oil dispersion procedure 1 Placea small amount of sample in a sample vial and add the appropriate oil 2 Shake the sample ideally using a vortex mixer to disperse the particles Some times ultrasonication is also required to disperse loose agglomerates 3 Once the particles are dispersed in the oil place a few drops of the dispersed sample on a clean microscope slide and place a cover slip B on top This encourages the oil to spread out Morphologi G3 Page 4 9 Chapter 4 Page 4 10 Preparing the sample ill 8140 Oil dispersion guidelines Note the following It is important to keep the layer of oil as thin as possible It may be necessary to leave the sample to settle for a few minutes before performing a measurement If the sample is too polydisperse and the oil rather viscous fine particles may be held at different heights within the oil layer and thus at different positions in the objective s depth of field This diagram shows particles at different heights in the oil layer with large particles holding the cover slip up ill 8136 This means many fine particles may be out of focus making measurements difficult Sometimes this can be overcome by using a less viscous oil and allow ing particles to settle to bottom of the oil layer before performing a measure ment If the particles are transparent and the refractive indexes of the particles a
171. pter 2 What is the Morphologi G3 Page e c4 MAM 0410 Hardware features Introduction This chapter introduces the hardware components of the instrument It covers System configurations describes the two types of system available Key components a summary of the key hardware components The instrument describes the main parts Polariser and Analyser options describes these options The 1X Adapter Kit option describes this option The software package is described in the following chapter System configurations Morphologi G3 There are two types of system B G3S this has an integral Sample Dispersion Unit SDU for automatic sam ple dispersion but can also be used with manual dispersion onto microscope slides etc WB G3 this has no integral SDU The user can use a separate sample preparation device or some other sample dispersion unit or prepare samples manually Page 3 1 Chapter 3 Hardware features Key components This diagram shows the key components of a typical system ill 8119 These are D e amp e The Morphologi G3S G3 instrument optical unit Integral SDU on the Morphologi G3S only 40V power supply for control electronics A computer running the Morphologi software and connected to two monitors We recommend that this computer runs the Morphologi software only Two flat panel monitors one for the software display and one for the live video feed
172. ptional sample plates B During the running of the SOP the user is prompted to select which slide posi tions to measure Click on slide number 1 and or 2 as required click again to de select The slide colour changes to green in the dialogue box as shown Pre measurement Slide s Select the slide positions to measure There are 1 slides required by the SOP Sample name Comments Select the first slide to use by clicking on it Then type the Sample name for this slide repeat this for the other slide The SOP s Measurement control settings specify how this produces the result If multiple slides are selected for example this can produce a record for each slide or one combined record Click OK MAM 0410 Optimising threshold settings Introduction In some circumstances the user may want to experiment with a number of thresh old settings in order to see the effect on the distribution rather than a few particles Whilst it is possible to perform several scans of the sample each with a different threshold a much faster way is to perform a single measurement with Save meas urement frames turned on and then use the Measurement from file feature to analyse the stored frames with different thresholds This Appendix explains how to do this Morphologi G3 Perform a scan to obtain the frames 1 Create an SOP in the normal way with the nominal settings for the sample Then use File
173. r of particles for example to achieve statistical significance set this here If merging multiple optics after the minimum number is reached measure ment will continue until all optics finish This is to prevent bias towards the first optic s used During the analysis the software will prompt for fur ther samples to be supplied until the specified requirement is met m Save measurement frames saved images can be useful for later analy sis purposes such as speeding up the process of setting the correct thresh old for a measurement see Appendix E To save all frames analysed during a measurement as JPG files select this check box and then browse to an appropriate directory create a new directory to prevent the system overwriting any previous image files Depending on the objective and scan area selected there could be hundreds of images generated per complete scan with each file totalling about 500KB Ensure that there is enough available space on the destination drive to accommodate the files Click the button when finished Morphologi G3 Page 6 9 Chapter 6 Measurement tutorial 6 The Sample carrier dialogue is displayed SOP Editor New SOP File Edit Help Qs Qi gli A Sample details Pre measurement settings Measurement control Sample carrier ce Sample Dispersion Unit Illumination Select the type of plate that will be used Optics selection Analysis settings Filters Classification Post me
174. r 5 Morphologi G3 Records tab The Records tab shows the user configurable tabular format shown in the exam ple below This has a column for each parameter measured It lists all the individual measurements in the opened measurement file using one record one row per measurement The record displays summary statistics for all the particles in the measurement 7 Morphologi Demo 19 9 07 Record 3 Sample 2 3 File Edit View Measure Tools Security Window Help D a Bia b d i Malvern Default E i ey Gy bey 2k A B Records Scattergrarr z Comparison Vel CE Diam M acy CE Diam Vol M mmm HS Circularity M T Aspect Ratio M gt Record ema Name TI a Particles CE Diameter Mean pm HS Circularity Mean Aspect Ratio Mean Elongal S 2 T Sample T September 2007 17 02 40 28 50 637 a a eT NN 4 Sample 3 03 October 2007 11 52 13 False 5 Sample 4 03 October 2007 15 00 54 False ES i i Mean 1 3 Std Dev RSD Minimum Maximum lt i mdyson By default records appear in the order that they were measured To sort records on a particular parameter click on its header field To toggle the order between ascending and descending order click again on the column header Right click on a record to display the menu of commands which apply to it These include Cut Copy Paste Delete Export Batch Print and Extract SOP To select multiple records use the standard Windows Ctrl and Sh
175. r Axis the graphic below shows the Major Axis and the minor axis The parameter is the angle of the Major Axis from a horizontal line The Major Axis also termed the orientation and minor axis are reported in degrees and can take values between 0 and 180 In the example below the Major Axis 95 l I ill 7687 The Major Axis passes through the centre of mass of the object at an orienta tion corresponding to the minimum rotational energy of the shape The minor axis passes through the centre of mass at right angles to the Major Axis Page e 8 MAM 0410 What is the Morphologi G3 Chapter 2e B Length all possible lines between two points on the perimeter are projected onto the Major Axis of the particle as shown below The longest of these projections is the length O of the particle l l I ill 7692 B Width all possible lines between two points on the perimeter are projected onto the minor axis D of the particle as shown below The longest of these projections is the Width ill 7691 B Aspect Ratio this is the ratio of the Width to the Length of the particle Width and Length are defined above It is calculated as Width Aspect Ratio Length Aspect Ratio values are in the range 0 to 1 A rod for example would typi cally have a low Aspect Ratio m Elongation this is 1 Aspect Ratio It is calculated as Width Length Elon
176. r Selection Morphologi G3 Page 8 5 Chapter 8 Viewing the results Particles shown in green If after selecting an area based on two parameters the user changes one of the selected parameters the particles which the original parameter had selected but the new one does not are shown in green Area Pixels b CE Diameter ym v Area Pixels Perimeter um he 10 100 1 000 10 000 100 00 1 000 10 000 100 001 1 10 100 1 000 1 000 000 100 000 Area uf a Area pr MI In the above example in the scattergram on the left the user selected CE diameter as a parameter They then changed this to Perimeter and selected some particles the scattergram on the right shows in green the particles which the CE diameter parameter would have included but Perimeter does not Viewing particles the Particles pane Double clicking on a particle in the Particles pane highlights it shows a zoomed copy of the image and displays its morphological parameter values in a Particle Details window as shown here An alternative is to right click on the particle and select Particle Details To zoom in or out on the Particles pane use the slider at the base of the pane Zoom Ol Page 8 6 Farticle Details Field Area Pixels Area uim Aspeck Ratio CE Diameter Lm Centre X Position m Centre Y Position um Circularity Convexiby Elongation H5 Circularity Id
177. r except Between type in one value to complete the filter For Between type in two values to define a range The and symbols for values allow successive ranges which do not overlap to be set up The Copy and Paste buttons can be used to copy filters between two SOPs When all filters are complete click the button 17 The Classification dialogue is displayed SOP Editor New SOP File Edit Help Qe Qig i 9 8 Blox Optics selection aaa Sample details Pre measurement settings Measurement control Classification Sample carrier Illumination 5x 6 5um 420um Parameter Operator Value 1 Value 2 Threshold Scan areas 10x 3 5um 210m Threshold Scan areas Analysis settings Filters Post measurement settings Reports Use this to set up any required classes as defined in Chapter 2 Class rules use AND logic meaning that a particle is only a member of the class if it meets all of the rules set up Particles can be a member of more than one class Q Note It may be better to make the first measurement without defining classes view the results then define classes using the Scattergram tab see Chapter 8 and add these to the SOP Future measurements made with the SOP will then have the classes set up The Copy and Paste buttons can be used to copy classes between two SOPs In the example below a class named Class A is defined using the Circularity and Elongation parameters MAM 0410 Measurement tu
178. ra tion or mechanical offset position etc Z Position Control focus controls the stage s vertical movement First set the size of increments by dragging the Step Size slider Next click on the up down buttons to move the objective Each click moves the objective by the step size The Z Position panel shows the current distance from the Z origin Z Stacking See Chapter 2 for overview information on Z Stacking 1 Select an appropriate point of focus Z Position on the image Z Stacking scans are made from the bottom up so for best results focus on the bottom of the particle For example if focus points can be obtained on the par ticle throughout the Z Position range 2 592mm 2 596mm select the lower 2 592mm Z Position before Z Stacking If it is hard to decide where the point of focus should be it can be helpful to focus on the surface of the plate except when using the Wet Cell or a prepara tion with a coverslip To do this switch to top light and focus the cross hair on the surface of the plate Select the number of images to use in the stack each stack is two depths of field Use the up down arrows or directly enter the number between 1 and 5 is anormal range The number entered includes the initial image taken at the point of focus It is best to experiment with the process to see how many images suit the view under analysis Start with low numbers such as 3 and work upwards until the optimal image is
179. rds tab and moved via drag and drop directly into another open application for example Excel The sec ond application must support tab or comma separated formats B A graph can be dragged and dropped into another application by holding the Ctrl key selecting the item and dragging it to the application B A graph can be copied by selecting it then using Edit Copy to copy it to the Windows clipboard It can then be pasted into another application All these methods are described in detail in this chapter Page 9 2 MAM 0410 Exporting results Chapter 9 Direct copying of data These two methods use standard Windows techniques Drag and drop Exporting data using drag and drop is as the name suggests simply a matter of selecting the record s dragging them to the target application and then dropping them Do the following 1 Select a record then move the cursor slightly until a small rectangle and cross appear 2 Press the mouse button down and drag the selected record s to the other application 3 Release the button All the parameters shown in the Records tab will be exported The same technique can be used to export graphs from a report Copying with the clipboard Records or graphs but not Scattergram or Comparison tab components can be copied by selecting them then using Edit Copy to copy the selection to the Win dows clipboard The item can be pasted into another application using Edit Paste
180. re shown below ill 8123 These are Dispersion chamber the sample is placed in the top of this chamber between two rings of aluminium foil in a sample cartridge Remove the cham ber from the Morphologi to insert a sample When the SOP or the command Measure Disperse Sample is run the gas air injection breaks the foil and disperses the sample under pressure Clamp holds the dispersion chamber in place Push the chamber firmly into P P P y position in this clamp Cap holds the sample cartridge in place Page 3 9 Chapter 3 Page 3 10 Hardware features Tubing release button push this in when inserting or releasing the air tub ing Air dispersant gas supply push this tubing down onto the top of the dis persion chamber Precision XY stage The stage holds the sample plate holder It moves automatically in the X and Y axes ill 7403 The components of the stage are X drive controls movement of the slide in the X left right axis The drive is controlled by the software or the joystick Warning A The front right hand side of the stage may become hot do not touch it Sample plate or Sample plate holder these are mounted on the stage They are described below Y drive controls movement of the slide in the Y backward forward axis The drive is controlled by the software or the joystick Note The XY stage may make a noise even when it is stationary
181. reached for the purpose Click Perform Z Stack Page 5 22 MAM 0410 Software features Morphologi G3 Chapter 5 The system now generates the required images and provides a composite view when complete View Z stack 2 stacked image Source view 16 Use the amp and Q buttons to zoom in an out of the view Click and drag the blue shaded area in the Source view panel to move around the image To save the Z stacked image click Save Image and select a relevant location images can be saved in either JPG or BMP format Other controls Fit to window check box this option selected by default allows the user to navigate the preview image smoothly in order to identify a region of interest on the sample Once centred the region of interest in the preview de select Fit to window This provides a closer view of the sample making it easier to obtain a more precise level of focus Reset button if this pressed the stage moves backward and uses the Z limit switch see Chapter 3 to re reference the Z position so the pinhole is in focus The scale bar below the image view indicates the scale SOP mode When the window opens as part of SOP creation see Chapter 6 The Grab button replaces the standard OK button see above If this button is unavailable click the Light calibration button Do not try to change the objective in the Optics pane the objective selected in the SOP Editor is the only one whi
182. report s select the Acrobat PDF Reports check box Select the check box for each report type required To produce a pdf file CFR 21 part 11 must be enabled 20 When all parameters are set up click OK When prompted to save the new SOP give it a name which will make it easy for operators to identify what it 1s to be used for It will be saved as a vsop file The SOP can now be run as described in the next section MAM 0410 Measurement tutorial Chapter 6 Editing an SOP A quick way to produce an SOP is to edit an existing SOP which was designed for a similar type of measurement To edit an existing SOP 1 Use File Open SOP or click i to open an existing SOP 2 Review the settings for each stage and make any required changes For details on any parameter refer to the above section Note The tree is an alternative way of stepping through the SOP stages and is quicker if the only changes needed are to one of the later stages 3 When all changes are complete click OK Note A copy of the SOP used is saved with each measurement and can be extracted for reuse by selecting the record and using Edit Extract SOP This ensures the original settings are preserved Morphologi G3 Page 6 27 Chapter 6 Measurement tutorial 3 Measuring the sample This section explains how to run an SOP like that created above If an integrated SDU is in use insert the charged sample chamber in the clamp before starting To
183. rm all service procedures that may require the removal of the covers Warning A Removal of the covers by unauthorised personnel will invalidate the war ranty of the instrument Unless advised within the content of this manual only Malvern Instru ments trained personnel are permitted to remove the main cover of any part Supervisor The supervisor is the person responsible for the management and safety of the instrument and its operation The supervisor is responsible for the training of the operators The supervisor can perform all user maintenance routines identified in the Essentials Manual Operator An operator is a person trained in the use of the system The operator can perform all user maintenance routines identified in the Essentials Manual Page 1 3 Chapter 1 Page 1 4 Introduction to this manual Warning Under no circumstances should the supervisor or an operator remove the main cover of the instrument Failure to follow these guidelines could result in exposure to hazardous voltages Assumed information To make full use of this manual the user should understand the following points Naming convention The Morphologi G3 is referred to either in full as the Morphologi G3 or as the instrument The combination of the Morphologi G3 instrument the computer and the Morphologi software is referred to as the system Menu commands Menu commands from the Morphologi software are always shown in bo
184. rompted to select which slide fil ter positions to measure Click on slide number 1 and or 2 as required The slide colour changes to green in the dialogue box as shown To deselect the slide click on it once more Page D 4 MAM 0410 Optional sample plates Appendix D Morphologi G3 Pre measurement Slide s Select the slide positions to measure There are 1 slides required by the SOP Sample name Comments Select the first slide to use by clicking on it Then type the Sample name for this slide repeat this for the other slide The SOP s Measurement control settings specify how this produces the result If multiple slides are selected for example this can produce a record for each slide or one combined record Click OK Recommended filters Filter diameter use 47mm or 25mm filters in accordance with the size of Filter Plate in use Filter grade The optical quality of filters is critical to the accuracy of results We recommend mixed cellulose ester filters due to the homogeneity of background they provide Whilst other papers are usable with the system they must meet or exceed the optical quality of mixed cellulose ester filters Filters of lower optical quality could result in darker areas of the filter being misinterpreted as particles by the system Also papers with grid lines are not compatible with Morphologi Suggested filter suppliers W Millipore http www millipore com B Whatman http
185. rt the Morphologi software 1 Power on the computer 2 Double click on the Morphologi fJ desktop icon to start the software Note If the icon is not on the desktop open the Windows Start menu and select Programs Malvern Instruments Morphologi to start the software 3 Ifsecurity is configured when the logon screen shown below is displayed type the user name and password if this 1s requested and click OK Chapter 10 shows supervisors how to administer security and set up users and groups User name 44 Type your user name to lag on Malvern usw 4 The software opens If a measurement file was open in the previous session this will automatically reopen If this 1s the first use of the software the Example Results vmes file will open Page 7 e MAM 0410 Measurement from files Chapter 7 The SOP Editor Image file analysis means using the SOP Editor This section describes this tool Each SOP Editor dialogue has a Help button advising how to complete it The SOP Editor comprises several dialogues that can be stepped through using the and Q3 buttons Image File Analysis A tree structure like that shown here appears in the left hand side of each dialogue This offers a quick way of moving through the SOP Editor just click on any mage files i Sample details branch in the tree to move to that stage Threshold Analysis settings Filters Classification The other buttons on the SOP Editor toolbar are
186. rt time in the centre of the scan area 16 The microscope stage moves to the light calibration position and begins calcu lating the optimum light intensity The Status bar reports Setting the inten sity for the objective Morphologi G3 Page 6 33 Chapter 6 Measurement tutorial The following dialogue shows its progress towards the target intensity Set Intensity Target Intensity 80 00 Measured Intensity When the microscope reaches the target intensity it moves straight to the next step Note The calibration algorithm is relatively insensitive to focus errors The offset between gratings and sample position is saved by the software This can be obtained in future by selecting the measurement record and using the Export SOP command 17 The measurement starts It may take anything from two minutes to over an hour depending on the SOP settings The Status bar and Summary win dow should show the number of particles analysed increasing Page 6 34 MAM 0410 Measurement tutorial Chapter 6 ZS SOP Measurement general vsop Particles CE Diameter um Mean 25 14 CE Diameter uim Std Dev 22 03 10 Percentile 10 88 10 100 10000 CE Diameter um 50 Percentile 16 41 90 Percentile 43 51 E Diameter um v C volume Transformation Classifications long needles large oblates short needles large oblates Unclassified 0 10 20 30 40 50 60 70 80 90 100 Number of Particles Multivie
187. run the SOP measurement 1 Check that the microscope symbol in the Status bar is not crossed out 2 Use File New Measurement to open a measurement file for the results It this step is omitted the result is saved to the current measurement file 3 Select Measure SOP or click the 5g button In the Open dialogue select the newly created SOP and click Open 4 The Measurement Manager appears for a description of its features see Chapter 5 ZS SOP Measurement general vsop 4 gt HM Q Documentation Start Stop Particles 0 CE Diameter um Mean CE Diameter uim Std Dev 10 Percentile 50 Percentile 90 Percentile E Diameter um v C Volume Transformation Classifications Unclassified Number of Particles Multiview Result Video There is a short initialisation routine 5 When the Start button turns green and the Status bar says Press START to measure click the Start button Page 6 28 MAM 0410 Measurement tutorial Chapter 6 6 Ifthe SOP has any sample details and pre measurement instructions these are displayed by this Pre measurement dialogue Pre measurement rn ee ae eS Pre measurement Documentation Pre measurement The Pre measurement tab if it is present describes actions to perform before the measurement can proceed Follow these instructions If the SOP needs sample information or other comments from the operator type these in on the Documentation ta
188. s The Trend plots selection area is used for selecting the Y axes parameters The Y1 parameter column is used to select the axis on the left of the graph and Y2 to select the axis on the right To choose a parameter press the Add button select the appropriate parameter and press OK Either highlighting the chosen parameter and pressing the icon or double clicking the parameter will again display the Select a parameter dialogue Use the Against area to select the X axis parameter The Properties dialogue for the Scattergram allows the user to set which param eters to plot on both the X and Y axes General graph properties Use the Properties dialogue to alter other attributes of the graph B Display or Option depending on the graph type inserted Display or Option will appear in the Properties dialogue B The Options tab or selection box in the above Display tab used to set the position of the key A Graph Tips option enables popup tips flags showing data points on the graph to appear on the report but not on the screen shown in the Report Designer MAM 0410 Creating custom reports Chapter 13 m Axis settings for both the X and Y axes whether a logarithmic or linear axis is required and the axis scales defined or auto scaling Graticule or grid lines can also be shown on the graphs B Fonts the font style used for all annotations on the graph B Upper Lower limits sets upper and lo
189. s They are described below Page e 6 MAM 0410 What is the Morphologi G3 Chapter Z Morphological parameters used Morphologi G3 This section describes the morphological parameters which the software uses to describe shapes The parameters are B Id as objects are detected during the scan they are assigned this as a unique identifier It represents the order in which they were found Each frame is scanned vertically and successive frames are acquired in the Y direction then in the X direction Id 1 N where N total particles found WB Centre X or Y position the position of the object s centre of mass relative to the nominal front left side of the slide This is slightly different from the physical border of the slide due to the extra clearance built into the scanning software Distances are reported in um 9 gt X gt To identify the actual physical position of the particle 1 Select the particle in the Scattergram tab ill 7697 2 Use the Microscope Manager button or the joystick to move to the Centre X Y coordinate 3 Switch to Episcopic illumination and use the focus target to project a white cross onto the slide The centre of the cross is the XY coordinate Page e 7 Chapter 2 What is the Morphologi G3 WB Area pixels Area microns the area is the visual projected area of the particle This is reported in both pixels and in microns um ill 7693 m Majo
190. s 12 1 Workspaces supplied by Malvern eee eee eee m IR 12 2 The cument WorKSDACGe 5 235 x X Aoc CES MOOR AE eA eo nC 12 3 Creating workspaces 0 ee es 12 3 EqitiNG a WOrkSpaGe 4 4 244 sees ae ee Ea KE X ap data eR oa 12 6 Deleting a workspace s s s ssar maaa 12 6 Morphologi G3 Page iii Table of Contents Morphologi G3 Creating custom reports IbtFOGIU CHLOE S sargia edo 800 5 UE do RAS ae A e e D od de de 13 1 USED US G Aiton nthe M a ge eei bra a a SH a dra he Gu a Seduta niat 13 1 Opening an existing report lesen 13 3 Creating anew report hh hr 13 3 Adding elements to the report voeux rp Go drea e se eed onc 13 4 Setting up the report elements 4 a diode CY C GE eR Y 13 6 Selecting CLEMENTS giu de eos b ta RNC UE ARE edt Se EX RR 13 14 Aligning and sizing elements leeren 13 15 Savilg eFeporbea kis BARES Le eee eee ee NUS ADSENSE 13 15 Viewing the new report 2 ee ees 13 16 Other information shown on a report seen 13 16 Protecting QTCDOM Es qna ce arci agri Eod ORAE RACE ante RR we Ge 13 16 Part 3 Appendices Specification SPeCIICAONS xa sc b ed So aa ee la ee a ee aoa ew od A 2 Minimum computer specification 0 ee es A 3 Chemical compatibility s eno ae OUR c UE HER e ai c Sed A 4 Unpacking instructions Regulatory statements CE Declaration of Conformity es C 1 PCG Notice US Only uet Qupd od sr desine e one ode Ru dee oaa NE X RE
191. s into the Value 1 and Value 2 fields 5 When the class is defined correctly rename it as required Over the default name Class 2 in the above example type a name for the class which makes clear what the particles are 6 Select the Exclude unclassified particles check box if the measurement is to include only the particles which are members of at least one class Classes can also be set up by B Using the Selection tab selecting particles then clicking the Save as Class button B Clicking the New button then selecting operators and typing values in the Classification tab This takes longer than the procedure outlined above it is the same process described for the SOP Editor s Classification dialogue in Chapter 6 Page 8 10 MAM 0410 Viewing the results Chapter 8 Morphologi G3 Viewing a particle as seen in the original image A particle can be viewed in its position in the sample In a sample on the plate for this the microscope must be connected Select the particle and click the EA button This opens the Microscope Manager centred on the particle For a file analysis the particle in the original frame Right click the particle then select View in original image in the displayed menu This opens the Particle in image dialogue Particle in image Sample 1 Bmp Raw source image Source view 100 no T Raw O Threshold q Centre particle This shows the particle and any neighbouring par
192. s its own toolbar described in Chapter 8 Statistics bar This shows statistics for the records selected including mean standard deviations minimum and maximum values Page 5 14 MAM 0410 Software features Chapter 5 Morphologi G3 Status bar and 21 CFR part 11 icon The left hand side of this area is used to display status information and also the tooltips for example when the cursor is held over the toolbar or a menu option A tooltip explains what a feature can be used for The name of the logged on user appears near the centre of the status bar If the 21 CFR part 11 feature key is installed the 21 CFR 11 icon appears in yellow on the right of the status bar Double clicking the active icon displays the feature key number note that the 21 CFR part 11 option is not detailed in this manual The microscope symbol describes the instrument status as follows 3 Instrument connected A Instrument not connected 73 Instrument initialising Hover over the icon to show instrument information such as serial number and firmware versions an instrument must be connected to the computer Page 5 15 Chapter 5 Software Features The Measurement Manager When an SOP is run the Measurement Manager appears giving control of the measurement The display is generally the same for all measurement types Its components are described below n IE 2 X Stop Help Close Summary 3 0 Particles 1401 Major
193. s one of the supplied workspaces Chapter 12 gives full details on workspace management Batch printing To print reports for more than one record select the records right click and select Batch print Alternatively select File Batch print The Batch print dialogue appears requesting the report template to print Batch print Batch printing allows you to print a separate copy of a report For each selected record Select which report you would like to use Circt Particles M hal Select the required report and click OK This produces one copy of the report for each record Morphologi G3 Page 8 3 Chapter 8 Viewing the results The Scattergram tab With a record selected in the Records tab click the Scattergram tab to display images of particles in the sample The components of the tab are described briefly in Chapter 5 this section shows to use the tab Initially all particles are shown in the order they were detected This section shows how to refine the selection of particles displayed in various ways The new particle set produced can be saved as a new record if required This section covers 1 Selecting particles with the scattergram setting up the scattergram based on any two parameters then viewing particles within areas of it Creating filters to include exclude particles Defining classes of particle Viewing full particle details Viewing a particle as seen in the original image Selecti
194. s to outline any instructions for the operator on cleaning equipment health and safety etc These are displayed on screen for the operator when the measurement is com plete Click the button Page 6 25 Chapter 6 Page 6 26 SOP Editor New SOP File Edit Help peack 1 A Sample details Pre measurement settings Measurement control Sample carrier Sample Dispersion Unit Illumination Optics selection 5x 6 5um 420um Threshold Scan areas 10x 3 5um 210um Threshold Scan areas Analysis settings Filters Classification Post measurement settings Note 19 The Reports dialogue is displayed Report actions Print selected reports Available reports O Area um IM O Aspect Ratio M O CE Diam M C CE Diam vol M CI CE Diam Circ Classification M C CE Diam HS Circ M O CE Diam Particles M C Circ Particles M O Circularity IM O Classification chart M O Classification table M O convexity M O Convexity HS Circ M O Elongation M C Histogram Undersize Distribution M O Hs Circularity M O Intensity Mean IM O Intensity SD M O Length M O Max Dist M O Multi record Statistics M O Particles M Measurement tutorial To print out the selected reports automatically after a measurement select the Print selected reports check box All reports available for the workspace are listed To produce a pdf file containing the
195. se left right This can be changed to suit personal preferences In one directions setting the stage moves left when the software joystick is moved left in the other the image moves left Reverse back This can be changed to suit personal preferences In one ward forward setting the stage moves forward when the software joy directions stick is moved left in the other the image moves forward Help menu This gives more details on how to use the Microscope Manager Use of folders The default locations for all files are sub folders of this folder C Documents and Settings All Users Shared Documents Malvern Instruments Morphologi The files are held in the following folders Type Extension Folder default Audit trail adt Audit Trails Deleted record del Result Data Export data txt Export data Export template edf Export Templates Image img Particle Data Measurement file vmes Result Data Report file vrep Report Pages SOP file vsop SOP Morphologi G3 Page 5 e5 Chapter 5 Software features Backing up and Archiving the system We recommend that files are copied moved from the computer regularly as part of two similar processes B Backing up the regular process of copying data to a server or other media as insurance against computer disk failure other hardware problems or problems in the laboratory This should be undertaken at a frequency probably weekly or monthly in line with compan
196. stick to mean move the stage leftwards or alternatively move the image leftwards MAM 0410 Hardware features Chapter 3 Tensioners Ensure the sliding tensioners are fully home and latched to hold the joystick verti cally in the off position In this position there is no power to the stage motors and the stage does not move Hot keys stage and focus motor speed The hot key on the left of the joystick adjusts the maximum speed of the stage This affects the X and Y axes equally The hot key on the right of the joystick does the same for the focus motor Z axis Hot key presses work as follows assuming that the starting position is maximum speed m First press reduces the speed to 25 of the maximum B Second press increases the speed to 50 of the maximum W Third press returns the speed to the maximum Digipot fine focus control The digipot knob on the right hand side of the joystick controls the focus motor Z axis Note that the movement is not proportional Note When the unit is being controlled by the Morphologi software only change the focus by using the software or the digipot Computer and monitors A computer of suitable specification is normally supplied For information on the computer requirements contact Malvern Instruments Polariser and Analyser options Morphologi G3 The Polariser and Analyser options can be used for episcopic microscopy The two options which take t
197. supervisor tasks configuring security managing Stand ard Operating Procedures SOPs managing workspaces and producing cus tom reports This user manual is a companion to the Morphologi Essentials Manual which gives Health and Safety maintenance troubleshooting and other vital information which all users must read Page 1 1 Chapter 1 Introduction to this manual How to use this manual We recommend reading this manual fully before starting the first measurement Those who are more familiar with particle characterisation instruments can jump straight to Chapter 4 for sample preparation Chapter 6 for a practical tutorial on making measurements or Chapter 7 for image file analysis The manual is divided into the following three sections Part 1 Operator s guide The Operator s guide contains all the information required by an operator using the Morphologi G3 Topics covered are W What the Morphologi G3 does The hardware components Preparing a sample for dispersion The software components A tutorial the steps to making a measurement including creation of Standard Operating Procedures SOPs Analysis of image files how to create an SOP to measure image files Viewing the results This shows how to interpret these in detail and how to compare measurements and how to modify results using filters and classes It also describes the reports produced and how to modify the format of the graphs in these rep
198. t on its side will damage the covers Posez toujours l instrument sur ses pieds Si vous le posez sur le c t vous endommagerez les couvercles Das Ger t stets auf seinen F en aufstellen Ein seitliches Aufliegen besch digt die Abdeckungen Se debe colocar siempre el aparato sobre sus patas Depositarlo de costado danaria las cubiertas Coloque sempre o instrumento na vertical sobre OS seus p s Se o colocar de lado ir danificar as tampas 10 Retain all packaging in case the instrument requires transporting in the future Conservez tous les emballages au cas o l instrument devrait tre transport ult rieurement Das gesamte Verpackungsmaterial aufbewahren falls in Zukunft ein weiterer Transport des Ger ts erforderlich wird Conservar todo el material de embalaje por si el aparato debiera ser transportado en el futuro Guarde todo o material de embalagem caso seja necess rio no futuro transportar o instrumento Morphologi G3 Page B 5 Appendix B Unpacking instructions Page B 6 MAM 0410 Regulatory statements CE Declaration of Conformity The CE badge on this product signifies conformance to European Commission Directives Morphologi G3 Page C 1 Appendix C Regulatory statements FCC Notice US only The Federal Communications Commission FCC mark on this product signifies conformance to FCC regulations relating to Radio Frequency Devices These have been satisfied by testi
199. tarts Sample carrier Sample Dispersion Unit Illumination Optics selection 20x 1 8m 100um Threshold Scan areas Analysis settings Filters Classification Post measurement settings Reports If operators need instructions to follow at the SOP start for example on how to prepare the sample select the Show these instructions check box and type in the instruction text below it These instructions will pop up on the screen when an operator runs the SOP Click the 3 button MAM 0410 Measurement tutorial Chapter 6 5 The Measurement control dialogue is displayed SOP Editor New SOP File Edit Help Quee e gdliiJ ii Sample details ent Pre measurement settings Measurement Control Sample carrier Sample Dispersion Unit S Fixed number of slides plates Illumination Optics selection 1 Analysis settings Filters Classification Slides plates combined into single result Post measurement settings Reports O A new result For each slide plate O Minimum number of particles Save measurement frames Specify one of the following B Fixed number of slides plates if the sample preparation method requires that the sample is distributed over four slides for example specity 4 here to ensure that the operator includes all four slides in the analysis Specify whether to combine or separate the results from these slides B Minimum number of particles if the measurement requires a numbe
200. ter shows how to measure images stored in a file For this there is no need for a microscope to be connected to the computer Files of images in any standard graphics format like bmp files can be measured For example a set of images produced using another microscope could be emailed to a Morphologi G3 user for analysis Notes Por reasons of software protection the PC must be attached to a Mor phologi instrument to use this facility Images in compressed formats like jpeg should not be used as some details may be lost during the compression This chapter covers W Getting started starting the software and placing the sample B The SOP Editor a brief overview of the tool used for image analysis B Measuring an image file use this procedure for all other image files After making the measurement m Particles can be sorted filtered and classified based on any morphological parameter and new records created using the filtered data By viewing both the particle images and the distribution it may be obvious that certain particles need to be excluded for example dust particles or fibres View and edit the results as described in Chapter 8 W Use the supplied reports to display distribution data and result statistics The Report Designer can be used to add more reports with different contents see Chapter 13 Page 7 1 Chapter 7 Measurement from Files Getting started The first step is to start the software gt To sta
201. the following options of the Layout toolbar or menu to align size them Button Function c Layout Align Align left im Layout Align Align right eed Layout Align Align top es Layout Align Align bottom Layout Make Same Width Fl Layout Make Same Height zn Layout Make Same Size The last element selected highlighted with eight dark squares provides the ref erence position for alignment The Layout menu also has commands to B Space Evenly several elements and Centre an element E Send to front or Send to back the selected element when two or more ele ments are overlaid Note Q Right clicking on a selected element also brings up the layout options Saving a report To save a report select File Save and save it in the Report Pages directory It must be in this location if it is to be available for inclusion in a workspace Reports are saved with the default file extension vrep If the report was designed solely in Screen Layout view a prompt appears advis ing that the Print Layout version is blank In Screen Layout view select Edit Select All and then Edit Copy Use View Page Layout and then Edit Paste to copy the content into the printed report page If the printed layout is to be different from the Screen Layout version make the changes here All that is then required is some rearranging and formatting to line up all the elements Page 13 16 MAM 0410 Creating custom reports Chapter 13
202. the main window and its tabs These are the Records tab Scattergram tab Comparison tab and the multiple Report tabs This shows what the software looks like at startup and after a measurement For more detailed information on using the tabs to view interpret results see Chapter 8 Software components describes the menus toolbar and status bar The Measurement Manager describes the tool used to make measure ments The Microscope Manager describes the microscope control software Use of folders describes the locations used for files Backing up and archiving the system A complete measurement tutorial is given in Chapter 6 Measurement of files is described in Chapter 7 Page 5 1 Chapter 5 Software features The main window This section shows what the main window looks like at different stages It describes the types of tab which can be selected using their tabs or the View menu Initial display When the Morphologi software is launched the Records tab is displayed 77 Morphologi Demo 19 9 07 Record 5 Sample 4 File Edit View Measure Tools Security Window Help D elg A BS i Malvern Default T GG r 2 Ra Bie ee Records i Scattergrar J Comparison VE CE Diam M ac CE Diam Vol IM E HS Cicularity E Aspect Ratio m i Elongation M Record Sample Name Date Edited Particles CE Diameter Mean um HS Circularity Mean Aspect Ratio Mean Elongation Mean Solidity M
203. ticle Z position 4 3 This image shows a fibrous particle that is standing off the slide surface position 1 If a single scan was taken at Z position 4 only the top part of the particle would be in focus and the rest would be nearly invisible which would be misrepresenta tive of the actual particle shape Z stacking enables the system to take images at positions 1 4 and then merge them providing a truer image of the particle The illustration below provides an actual comparison of how Z stacking can improve the overall clarity of an image Image A shows a standard particle scan and displays a level of blurring on some parts of the fibrous particle Image B shows the same particle after Z Stacking and exhibits a good degree of overall focus e 3 t E g T a m lt e a Chapter 5 contains details on how to perform a Z stack from within Microscope Manager Chapter 6 provides information on how Z stacking can be implemented in an SOP MAM 0410 What is the Morphologi G3 Chapter e Filters The user will often want to exclude some particles on the plate from the analysis These include the following WB Contaminants these may include particles of dust or fibres If any of these have consistent properties filters can be set up to exclude them from the analy sis For example dust may be excluded based on its size Area or fibres based on their Elongation B Particles which it is known will
204. ticles The small rectangle in the Source view panel shows the location of the selected particle in the origi nal image This rectangle can be dragged round in the Source View panel after doing this click Centre Particle to return the view to the originally selected particle Select Raw to view the original image or Threshold to see it with the thresh old applied Note Once the dialogue is open another particle can be selected in the Parti cles pane and View in original image selected again there s no need to open and close the dialogue each time Page 8 11 Chapter 8 Page 8 12 Viewing the results Viewing a particle within the scan area Scan Area Composite provides an image of the overall scan area in which a parti cle was originally recorded allowing the user to examine the original context more easily effectively a zoom out option The system creates this composite view by tiling together all the individual frames taken by a particular measurement to assemble a single image of the whole scan area This is particularly helpful for identifying flaws on the carrier that could affect measurements For example when used in conjunction with the optional filter plates see Appen dix D this function could help to detect a stained or imperfect filter that might otherwise have gone unnoticed under high magnification analysis Additionally the Save Image facility can be used to provide a record of the entire filter
205. time the system is run a single user named Administrator and one user group named Administrators are created automatically The Administra tors group is originally set to only allow configuration of the security system and to deny access to all other features of the system The first task is to set up the Administrator account To set up the administrator 1 Select Security Configure security to display this dialogue Security Configuration User Options Help Username Full Mame Description AS Administrator Administrator Built in account For administering security Groups Description E Administrators Built in account for administering security When the software is run for the first time the security system defaults to a member of the Administrators group with no password 2 Select the Administrator name in the first row and then User Properties Confirm the blank password for the Administrator account that 1s do not type anything in then click OK 3 Click Options Security settings and in the Security Settings dialogue shown below select the Enable security check box and click OK Security Settings Morphologi G3 Page 10 3 Chapter 10 Security Security is now enabled on the system This forces subsequent users to log in to access the software Note If the 21 CFR part 11 feature key is installed a different dialogue is dis played Once 21 CFR part 11 security is enabled it cannot be disabled
206. tings C Configure ER ES settings C Configure user workspace settings C Control display of status bar C Control display of toolbars C Create and edit export templates C Create and edit reports Create and edit SOPs Enter a Group name and a Description of the group s purpose Example names might be B Operators general users of the system B Supervisors skilled users performing configuration and creating SOPs The Members list shows all the users currently assigned to the group To add a user click Add a list of all users not currently allocated to that group 1s shown Select one or more users hold down the Ctrl key to select multiple users and click OK to add those users to the group If no users have been added yet add these as described below then add them to the group Note To remove a user from a group select them in the Members list and click Remove This removes the user from that group not from the system 2 Use the Permissions list to enable disable functions of the software for the group Simply check the boxes for the permissions required 3 Click OK to save the group To edit an existing group 1 Double click on an existing group in the Security Configuration dialogue to display the Group Properties dialogue 2 Proceed as described above for adding a group Morphologi G3 Page 10 5 Chapter 10 Security Setting up individual users Each user is defined by their WB Uniqu
207. torial Chapter 6 Morphologi G3 Classification 18 Parameter Operator Value 1 Value 2 i Class A gt Exclude class Circularity 0 600 Elongation Between 0 500 0 850 lt select zselect To add a class first click the New button and type in a name for the class over the text lt Classname gt Next click on the Parameter list and select a parameter In the Operator list select gt gt lt or Between For any operator except Between type in one value to complete this part of the definition For Between type in two values to define a range The and ES symbols for values allow successive ranges which do not overlap to be set up To exclude a class from the analysis or to use AND logic to exclude particles select its Exclude class check box This acts as a more complex means of excluding particles than the Filters dialogue allows as multiple parameters can be used to define the class Finally at the bottom of the dialogue select the Exclude unclassified parti cles check box if the measurement is to include only the particles which are members of classes We recommend leaving this unchecked during method development if this results in a high percentage of unclassified particles it sug gests that the classes need tuning Once the refined class definitions are correct this check box can be selected Click the G9 button The Post measurement settings dialogue is displayed Use thi
208. ts shown in a Report tab as shown in this table Workspace Classification and Particles Combined Reports Malvern Default Other reports Parameters Count and 96 for each class created CE Diameter Mean Circularity Mean HS Circularity Mean Convexity Mean CE Diameter Mean HS Circularity Mean Aspect Ratio Mean Elongation Mean Solidity Mean Convexity Mean Area Mean um Circularity Mean Length Mean Max Distance Mean SE Volume Mean Width Mean Reports Classification chart M Classification table M Particles M CE Diam Circ Classification M CE Diam HS Circ M CE Diam Particles M Circ Particles M Convexity HS Circ M Histogram Undersize Distribution M CE Diam M CE Diam Vol M HS Circularity M Aspect Ratio M Elongation M Solidity M Convexity M Intensity SD M Intensity Mean M Area M Circularity M Length M Max Dist M SE Vol M Width M If these are not suitable new workspaces can be created as described in this chapter Page le e MAM 0410 Workspace management Chapter le The current workspace The Workspace toolbar shows which workspace is currently in use In this example the Malvern Default workspace is being used Malvern Def aul Change between workspaces by clicking the list button on the toolbar and selecting the workspace to use from those listed This immediately updates the window to use the parameters and reports define
209. ubleshooting Installation this gives enough information to allow the user for example to move the system from one laboratory to another Instructions are also given on how to install the Malvern software if the user upgrades the computer system Help desk All queries regarding the system should initially be directed to the local Malvern Instruments representative Please quote the following information Model and serial number of the instrument located beneath the triangular panel on the right hand side of the instrument The software version to find this select the command Help About Morphologi in the software The firmware version to find this hover over the software s microscope icon Contact the United Kingdom help desk if the local Malvern Instruments repre sentative is not available Its direct line is 44 0 1684 891800 Note Q This help line is primarily English speaking Page 1 5 Chapter 1 Page 1 6 Introduction to this manual Remote support Malvern Instruments offers a remote support service delivered by an Internet con nection Benefits include fast and efficient fault diagnosis reducing downtime and costs On line user training is also available plus software updates A high speed Internet connection 1s recommended for making use of this facility Malvern Website www Malvern com The Malvern Instruments website offers a comprehensive range of particle charac terisation resour
210. uments Malvern Instruments Morphologi SOP The vsop files can be placed elsewhere on the C drive or network and accessed by browsing to this location Page 11 2 MAM 0410 Workspace management Introduction This chapter describes workspaces It covers How to use workspaces Workspaces supplied by Malvern Instruments The current workspace Creating workspaces Editing a workspace Deleting a workspace How to use workspaces Morphologi G3 A workspace specifies what is displayed on the screen in two ways WB Which parameters are shown in the Records tab m Which reports are produced and which Report tabs are available Use workspaces to WB Customise windows to show only the relevant parameters and reports m If required create a personalised user workspace showing only the parameters and reports relevant to the user Note i The current workspace view will apply to all open measurement files Page 12 1 Chapter 12 Workspace management Workspaces supplied by Malvern Four workspaces are supplied with the software These control the appearance of the software when it is initially installed in two ways B The parameters shown in the Records tab all the workspaces show the Record number Sample Name SOP name Date whether Edited or not and Number of particles in the first five columns The table below shows which other parameters each workspace includes B The combination of repor
211. us bar is not crossed out If it is check that the microscope is powered on and connected properly 2 Select Tools Microscope to open the Microscope Manager Microscope Image Instrument Joystick Help a Glass Plate 120 100mm Slide 1 x oe ae Optic A a Light Z Position Control Step Size um HE 69 07 8n Custom light calibration P E PN Standard light calibration Z Stacking 2 z Perform Z Stack x Y Position 2 Position 35 373 mm 81 575 mm 0 255mm c Move to preset position v Fit to window L 200 um 4 Note Q For an overview of this dialogue see Chapter 5 Morphologi G3 Page 6 3 Chapter 6 Page 6 4 Measurement tutorial In the Slide plate list on the toolbar specify which sample plate is in use Click the Move to preset position button and select its Centre of slide plate option Click the Optic button this is showing 2 5x in the above example and select the objective with the lowest available magnification Note Q Every time an objective is changed the light intensity must change too A 6 T 8 higher magnification objective requires more light and vice versa After each objective is calibrated for the first time the light intensity value is recorded for future use The Light calibration button is provided only for use in rare situations for example if the light intensity has been cali brated accidentally on a dark surface Practise moving arou
212. vailability etc so some way of characterising shape is required Shape factors circularity convexity and elongation Three commonly used shape factors are described below Circularity One way to measure shape is to quantify how close the shape is to a perfect circle Circularity is the ratio of the perimeter of a circle with the same area as the particle divided by the perimeter of the actual particle image Several definitions of circularity could be used but for accuracy the software reports HS Circularity HS for High Sensitivity in addition to circularity HS Circularity has a squared term in the numerator and denominator to sensitise the parameter to very subtle variations in the area perimeter relationship It is defined as follows HS Circularity 4xA P Where A is the particle area and P is its perimeter Circularity has values in the range 0 1 A perfect circle has a circularity of 1 while a spiky or irregular object has a circularity value closer to 0 Circularity is sensitive to both overall form and surface roughness The shapes below show how circularity 1s affected both by overall form and symmetry and by surface roughness Circularity 1 ERES Circularity 0 47 Circularity 0 89 P Circularity 0 52 Circularity 0 47 enn Circularity 0 21 ill 7659 ill 7660 Circularity is a good measure of what might be described as deviation from a perfect circle However it is important to remember that it is
213. value for each particle is shown This pane initially displays all particles but as filters are applied and classes defined some may be excluded A red X is used to show an excluded particle as shown for particles 3 and 4 above It can also display particles in selected areas of the scattergram only Zoom control drag the slider as required 2e Q J Q Particle Details window shows a zoomed image of the selected particle and also its parameter values Comparison tab The Comparison tab provides a powerful means of comparing all the morpholog ical distributions of multiple measurements Use it to Automatically find the morphological parameter that is varying the most between measurements See at a glance how all morphological parameters vary across a set of measure ments Automatically cluster measurements according to their similarity Establish how results cluster based on a chosen morphological parameter Analyse correlations between expected similarities and actual measurements Assist in determining morphological pass fail criteria These features are useful for Quickly finding the most significant morphological parameter separating eood and bad batches and setting pass fail criteria for that parameter Checking that a process is producing consistent product across all morphologi cal parameters Identifying which morphological parameters are important to a product Identifying vari
214. vember 2006 True 396 Each row record represents one measurement contained in this file Measurement file management depends on user preference For example m A separate measurement file can be used for each type of sample m A separate measurement file can be used for each week or month m A separate measurement file can be used by each user a One measurement file can be used for all the measurement records This is not recommended for all records but results which are to be overlayed or compared using the Comparison tab must be in a single file To find the SOP see below used to produce a record right click the record and select Extract SOP Note If more than one measurement file window is open the measurement record will be saved to the currently active file When the software starts it automatically opens the last measurement file used Page e 18 MAM 0410 What is the Morphologi G3 Chapter 2e Morphologi G3 Standard Operating Procedure SOP An SOP is like a template that defines all the measurement parameters and settings Using these ensures that measurements made on the same type of sample are made in a consistent way SOPs can be created or modified as required This is useful in Quality Control environments A single SOP file vsop can be emailed to differ ent sites where users need to measure the same product in the same way SOPs are also ideal for measuring the same sample in slightly different w
215. vern Instruments Morphologi G3 Page 5 13 Chapter 5 Software Features 3 Toolbars The toolbars are located below the menu bar They provide button shortcuts for performing the most popular operations Each button has an equivalent menu command For example clicking the amp button is the same as using the File Open menu command Toolbar appearance Toolbar and use Standard toolbar use this for file open E and print operations cut copy and paste undo and redo ZEE MEN Workspace toolbar use the list to select create or delete a workspace Use the but ton to view or change the properties of the current workspace Classification Particles SOP toolbar use this to create edit or t E be 2 of dA run an SOP The drop down lists next to buttons only show SOPs which have been run already ENT The final buttons open the Microscope Manager load or park the stage E Navigation toolbar use this to move up Pon and down a list of records Note the following W To identify a toolbar button s function move the cursor over it A tooltip is displayed and a short description appears in the status bar WB As with the menu bar if a tool is not available its button is shown greyed out W The content and appearance of the toolbar can be changed using the View Toolbars Customise command W Display of any toolbar can be turned on off using View Toolbars m The Scattergram tab ha
216. w Result Video If more than one objective is selected in the SOP the light intensity check step 16 is repeated for each objective If additional slides are required the user is prompted for these 18 Once the measurement sequence is complete any post measurement informa tion for the SOP is displayed The operator may need to enter documentation at this stage as in step 6 19 The Measurement Manager now prompts the user to click Start to make another measurement or Close to shut the measurement display and return to the main Morphologi G2 application The new record will be shown in the Records tab It will be the last record listed Morphologi G3 Page 6 35 Chapter 6 Measurement tutorial 4 Viewing and editing results All particles can be sorted filtered and classified in the Scattergram tab based on any morphological parameter and new records created if necessary using the fil tered data By viewing both the particle images and the distribution it may be obvi ous that certain particles need to be excluded for example if dust has settled on the sample plate View and edit the results as described in Chapter 8 5 Creating reports Reports are supplied to display distribution data and result statistics The Report Designer can be used to add more reports with different contents see Chapter 13 Page 6 36 MAM 0410 leesuirernerc from files Introduction Morphologi G3 This chap
217. ware features Sample plates Several sample plates are available as standard and other application specific plates will be available in future Always clean sample plates between samples The sample plate configuration depends on the instrument B G3S the SDU will only disperse sample onto an SDU glass plate The other sample plates can still be used for manual dispersion use of microscope slides etc WB G3 this can use two types of sample plate SDU glass plate G3s only The SDU disperses the sample evenly onto a circular area of this plate Always use this plate in the previous diagram for SDU dispersions G2 four slide plate This microscope slide holder can hold up to four slides and is useful for making several measurements sequentially For example reproducibility of sample prepa ration can be evaluated by measuring four slides producing a separate record for each and over plotting the results Alternatively if there are insufficient particles on one slide several slides can be combined into one result Each slide is held in place by a sprung clamp slide The four slide plate looks like this ill 8157 Note Q See also Appendix D Optional sample plates MAM 0410 Hardware features Chapter 3 Digital camera The FireWire digital camera system is under the removable part of the cover at the front of the main body The back of the camera has two connections and an LED Q cpe sS
218. wer warning or action limits Line styles Select Configure Line styles to display a line styles dialogue This enables each measurement result line in a graph to be either coloured increased in thickness or have its line style changed Select the Line style from the list and use the Symbols tab to alter the symbols used for data points as required Classification table A Classification table looks like the following example showing how many parti cles are in each class and optionally how many are unclassified Record Number Sample Name BEE Sample 7 Sample 7 Sample 7 sample 7 sample 27 To configure the display right click on it and select Properties Use the Proper ties tab which this shows to set up the display which can be arranged horizontally as shown above or vertically Selecting elements Morphologi G3 When selecting elements B To select one element click on it B Toselect multiple elements hold down the Shift or Ctrl key and click the mouse button on each element in turn Alternatively hold the left mouse but ton down and drag a box marquee over a group of elements m If one element is overlaid by another and cannot be selected select any element and use the Tab key to step through all the elements one by one until the cor rect element is selected Page 13 15 Chapter 13 Creating custom reports Aligning and sizing elements Select two or more elements and use
219. xcept for the second CE Diameter report both number and volume based Report name on tab Area um M Aspect Ratio M CE Diam M CE Diam Vol CE Diam Circ Classification M CE Diam HS Circ M CE Diam Particles M Circ Particles M Circularity M Classification chart M Classification table M Convexity M Convexity HS Circ M Elongation M Histogram Undersize Distribution M HS Circularity M Intensity Mean M Intensity SD M Length M Max Dist M Particles M SE Volume M Solidity M Width M Data shown Area um Aspect Ratio CE Diameter number based CE Diameter volume based CE Diameter and Circularity Also shows the class summary CE Diameter and HS Circularity CE Diameter plus particle images Circularity plus particle images Circularity A bar chart showing classes A summary of the classes Convexity Convexity and HS Circularity Elongation Shows these for CE diameter HS Circularity Mean Intensity Intensity standard deviation Length Max Distance Images of particles sorted by Id SE Volume Solidity Width The reports in each workspace are listed in Chapter 12 MAM 0410 Viewing the results Chapter 8 The reports available are specified on a workspace basis by the Report pages tab in the View Workspace Edit Workspace dialogue Two examples are shown here CE Diam HS Circ report This shows the CE Diameter and HS Circularity data 7 Morphologi
220. y procedures B Archiving the process of moving older data from the computer for example when the disk becomes nearly full Note Image files stored as img files in the folder Particle Data take up a lot of space When copied from the control computer we recommend that before storage these are compressed using a tool such as Winzip Follow the procedures described below Backups To allow recovery of the full system make copies of the following assuming that the software 1s installed on the C drive B The entire contents all sub folders of the folder C Documents and Settings All Users Shared Documents MMalvern Instruments Morphologi B From the following folder the files PlateDetails CFG SiteSettings CFG and any files with the suffix wrkspace C Documents and Settings All Users Application Data Malvern Instruments Morphologi Note Q The Application Data folder may be hidden To restore the files simply copy them back to their respective locations Page 5 26 MAM 0410 Software features Morphologi G3 Chapter 5 Archives For archiving of older information If the distribution data are sufficient and the individual image files are not required this will save significant amounts of space in the archive but will not permit re analysis of the data in future save the contents of all sub folders of the following folder except for Particle Data C Documents and Settings All Users Shared Do

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