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AIDA 1D TLC User's Manual

1. Using the Select Rows command you can choose four special row types to be displayed in the Result Table In the dialog box that appears click to select or deselect the appropriate options Clicking Total Sum and or Remainder inserts the respective row s into the table Separator displays an extra separator row showing the name of the profile Select Rows Ponls2 adf X Show Profile Rows OK Separator Cancel v Total Gammel jv Sum Ps es eet Figure 2 38 Select Rows Dialog Box Select Percent Reference This command allows you to set the reference value for the percentage calculation This can be either a distinguished peak defined by the Set to 100 command of the table contextual menu see above the value for the total profile or the Sum The later two are set via the following dialog box Percent Reference Pools2 adt Ts x Set 100 percent to Total profile C Peak summation OK Cancel Figure 2 39 Percent Reference Dialog Box The Change Profile Names command allows to re name the densitometer windows and resulting profiles according to your preferences The names will appear in all result tables and image menus Profile Name Fetraz adf Set name of 2 profile to Cancel Figure 2 40 Profile Name Dialog Box Using 1D TLC 4 raytest Quantity Calibration Window To open the Quantity Calibration window Choose Quantity Calibration from the View menu or
2. Password Conca Figure 1 2 AIDA Registration Login Dialog Box To activate the user registration in AIDA the first time you must enter a password for the system administrator account To enter the password for the system administrator account 1 Choose Create User Registration from the Options menu The AIDA Registration User Management Setup dialog box appears 2 Type a password for the system administrator account in the Password text field of the Aida Registration User Management Setup dialog box and confirm the new password by reentering it in the Confirm Password field Introduction 7 AIDA Registration User Management Setup X This dialog will activate the user management of this program From now on the program will require a user login an each start Please enter an administrator password for the user management Be sure nat to Forget this password Password E m Canfirm password enne Cancel Figure 1 3 Entering Administrator Password 3 Click OK to apply the changes Now the user registration 1s activated and only registered operators have access to the program When the user registration is activated and the operator has administrator privileges the User Registration command is selectable from the Options menu On choosing this option the AIDA Registration User Management dialog box appears allowing you to add new users or to modify or delete existing user accounts AIDA Regi
3. Select Profiles e TN Peak y Peak Typ 5 std 3 2 5 std 3 2 5 std 3 2 nsi Figure 2 26 Changing Column Width Using 1D TLC 33 Contextual Menu of the Profiles Window Graphics Pane Right clicking on the Profiles window s graphics pane opens a contextual menu providing you with commands for deleting creating peaks deleting profiles baselines as well as several display settings see below i a Delete Peaks Create Peaks Delete Profiles Delete Baselines Display k Multiple Data Smoothing Tile Scales wv 3d rid os Ter prolate Profile Overlays wv Extremal INE MEN Ins Change Profile Mames wv Show Baseline Hide Baseline Subtract Baseline Figure 2 27 Contextual Menu of the Profiles Window s Graphics Pane From the submenu of the Display command you can specify the appropriate display mode Multiple Tile and 3d explained earlier as well as how the data should be displayed You can choose between the Discrete display of data points pixels in the profile the display of Extremal data representing the maximum value for all points between the determined data points or the display of Interpolated curves between data points Further you can specify to show or hide the baseline using the Show Baseline or Hide Baseline command respectively In addition you can choose to subtract the baseline from the data by selecting Subtract Baseline 34 AI
4. Protocol Print Print Preview Audit Trail Format Portrait C Landscape Frint W Configuration complete lists C positive lists W Data Trailing Figure 2 84 Protocol Print Audit Trail Pane As for the Region Report you can select the paper orientation Portrait or Landscape by clicking the appropriate radio button in the Format pane The options in the Print pane enable or disable the printing of the Data Trail settings Configuration and or the recorded data from the Audit Trail window Data Trailing In addition if the Configuration option is checked you can choose to print the Complete Lists of settings or just the Positive Lists the checked options only 74 AIDA ID TLC User 5 Manual Print In AIDA there are two ways for printing your image and evaluation data e Choose Print from the File menu or Use the Protocol Print pane on the Publishing Layout toolbox The Print command of the File menu allows you to print your data on the fly without permanently storing the layout printing settings whereas the Protocol Print pane provides you with a complete set of printing features which includes saving the setting with the data file for reasons of reproducibility Choosing the Print from the File menu opens the Print dialog box allowing you to configure the printout to contain only the overlays results and graphs that are currently needed Click the checkboxes for the various it
5. 1 On the File menu click the Import menu item and then choose Templates from the submenu 2 In the Import dialog box that appears select the template to be imported and click Open xj Type OTF File Size 40 0 KB all File name pools2 att Files of type ADA Template Format L ancel Figure 2 95 Import Template Dialog Box The imported profiles can be arranged to fit the image by moving the whole array using the AIDA standard overlay editing functions Using ID TLC 81 Importing Quantity Calibration 1 To import the calibration curve from another image e g taken at the same time with the same BAS screen or from a second gel handled at the same time and captured under identical conditions use the Import command of the File menu The Quantity Calibration Import dialog box appears allowing you to specify the calibration file cal Quantity Calibration Import Ag test adf Type CAL File Size 255 Byte Ju File name Jag test cal Files of type ADA Calibration L ancel Figure 2 96 Quantity Calibration Import Dialog Box 82 AIDA ID TLC User s Manual Performing 1D TLC Evaluations 3 Setting the TLC Plate and the Origin Front Line The first thing to do when doing a 1D TLC evaluation is to set the TLC plate with the origin and front lines Consider the following image 5 mu Ges sin 5 E E p JFramnt Li JB HOM ot ps UG Originsine Figure 3 1 T
6. 98 AIDA ID TLC User s Manual Configuring the Evaluation Output Following the calibration process outlined above the entire image is calibrated and the results will be shown in the table pane of the Profiles window One or more profiles can be displayed at the same time as outlined in the Profiles Window part of this manual It is important to use the Retention Factor Normalization function to get peaks of identical size at comparable positions Wl Profiles Petra2 adf Intensity PSL Peak Type Rel Pos Integral Bkg Integral Bkg AF Value piel PSL PSL PSL 1 Profile 1 E 2 1302 15803 1280 14528 oor 3 tea trea 1083 16808 014 4 2240 15630 335 15755 019 5 2508 s 345 596 022 hee Figure 3 25 Retention FactorNormalization Example Performing 1D TLC Evaluations 99 100 AIDA ID TLC User s Manual
7. Clicking the Options button changes the content of the toolbox revealing options for the lane as well as the peak determination A Lane amp Peak Determination Lane Determination Options intensity summation Show profile after setting lane while dragging Peak Determination Options wam overlap cancel Figure 2 53 Lane amp Peak Determination Options Panel A profile can represent the Intensity Summation the Averaged Intensity or the Optical Density Select the appropriate option from the pull down menu of the Lane Determination Options section For applications with radioactive TLC usually Intensity Summation is used If the After Setting Lane option is checked the Profiles window opens directly after the densitometer window has been drawn If the While Dragging option is checked the profile of the currently dragged densitometer window is continuously updated in the miniaturized profile window of the toolbox The Peak Determination Options pull down menu is used to define AIDA s action if an overlap of peak regions is detected Peak Determination Options warm overlap hs allow overlap avoid overlap v warn overlap Figure 2 54 Peak Determination Options Pull Down Menu 52 AIDA ID TLC User s Manual Allow Overlap Avoid Overlap and Warn Overlap can be selected Avoid Overlap will disable the overlapping of regions by either keeping peaks separate or by unifying two strongly overlapp
8. Double click the miniaturized calibration curve display on the Quantity Calibration toolbox 4 Bl x Linear Regression y 13 893x 14 80 Correlation Coefficient r 0 9993 Intercept on x Axis 1 062 Integral Bkg QL ce c 0 10 20 30 40 50 60 70 60 390 100 120 14 Quantity ng Profile Peak Integral Bkg QL Quantity Std ng Quantity Recalc nq 1 Standa g 1402 9 100 0 99 66 11 Profili 1 494 6 34 44 Figure 2 41 Quantity Calibration Window Example The Quantity Calibration window is bipartite the upper pane displays the curve of the calibration function the underlying function the correlation coefficient and the x axis intercept The lower one contains a table listing the standards their given and recalculated standard values so you can easily evaluate their differences This can be useful to estimate the error for a certain amount calculated for an unknown band The graphical representation can be adapted using the contextual menu attached For a more detailed view of the plot you can activate the AIDA standard zoom functions described above The table can be exported as a text file or copied into the clipboard Additional columns can be displayed Just right click on the table heading and choose the Select Columns command from the contextual menu that appears The Columns Selection dialog box opens allowing you to select the desired columns from the list 42 AIDA ID TLC User s
9. p 0 D p 2 C A AIDA ID Thin Layer Chromatography User s Manual Part Number 2004 04 1 1 2004raytest IsotopenmeBgerate GmbH Benzstr 4 D 75334 Straubenhardt Germany All rights reserved This product or document is protected by copyright and distributed under licenses restricting its use copying distribution and decompilation No part of this product or document may be reproduced in any form by any means without prior written authorization of ray test and its licensors if any RESTRICTED RIGHTS LEGEND Use duplication or disclosure by the government is subject to restrictions as set forth in subparagraph c 1 11 ofthe Rights in Technical Data and Computer Software clause at DFARS 252 227 7013 and FAR 52 227 19 The product described in this manual may be protected by one or more U S patents foreign patents or pending applications TRADEMARKS AIDA AIDA Image Analyzer AIDA Annotation AIDA AIDA Array Metrix AIDA Array Compare AIDA Multi Labeling AIDA 1D Evaluation AIDA ID Densitometry AIDA 2D Densitometry AIDA Easy Array AIDA Whole Body Autoradiography AIDA 1D Thin Layer Chromatography and AIDA 2D Thin Layer Chromatography are trademarks or registered trademarks of ray test inthe United States and may be protected as trademark in other countries All other product service or company names mentioned herein are claimed as trademarks and trade names by their respective companie
10. this button Background Determination Bka from Regions T Please set bkg regions to define the background E Figure 2 59 Bakground Determination Section Choosing Bkg from Baseline from the Background Determination pull down menu changes the content of the Background Determination section to reveal the baseline determination controls Background Determination Bka from Baseline T Level smoothed pixel e Elia Figure 2 60 Baseline Determination Controls 4 Clicking this button activates the Edit Baseline function This button removes the baseline s of the currently selected profile s With this button the baseline can be determined automatically Use the Smoothed parameter to omit single pixel noise The Level of sensitivity can be set to various values reaching from 0 insensitive to 10 extremely sensitive aim Using 1D TLC 55 Peak Determination Section This part of the toolbox is used for the determination of peaks Feak Determination position by center d sensitivity FWHM pixel Figure 2 61 Peak Determination Section 3 A Use the pull down menu to mark the position of the band by Position by Maximum or Position by Center position by center EU position by maximum wv position by center Figure 2 62 Peak Determination Pull Down Menu This button is used for manual peak assignment Click this button to delete all currently selected pea
11. 1 On the AIDA Registration User Management dialog box click Modify to open the Modify User dialog box User name Full name Permissions Password Confirm password Comment Cancel Help Figure 1 6 Modifying Users 2 Edit the database entries you want to change 3 Click OK to confirm the changes If the operator has user permission the Change Password option instead of the User Registration option is selectable from the Options menu To change your password 1 Choose Change Password from the Options menu The Change AIDA Registration Password dialog box appears Change AIDA Registration Passwo rd X You can change the password of user doe Old password Mew password Confirm password 3 Cancel Figure 1 7 Changing Password 2 Type your current password in the Old password text box 3 Type your new password in the New password text field and confirm it by reentering it in the Confirm password field 4 Click OK to apply the changes 10 AIDA ID TLC User s Manual At the end of the session you can choose to log on as a different user instead of exiting the program Figure 1 8 Exit Aida Dialog Box Introduction 11 12 AIDA ID TLC User s Manual Using 1D TLC 2 Data Windows in 1D TLC In addition to the AIDA standard data windows Image and Histogram the following data windows are available in the 1D TLC mode Profile
12. Box Lanes Tab This opens the Columns Selection dialog box which provides you with a list of possible labels e g lanes can be labeled by Number Profile name or not at all Columns Selection I x Show Columns Hide Columns B Bkg GL Integral GL Recale Quantity Integral Bkg GL Integral 95 Type Nama Integral Bkg 95 Raeralc Quantity 95 uU Hidelegend Hide all Figure 2 7 Columns Selection Dialog Box Using 1D TLC 17 8 qe amp raytest After you have determined the type of labeling you can set the font font size and font style bold italic roman typeface on white background as well as the rotation 0 45 90 for the labels The detected lanes can be surrounded by a frame Border Lines and tracked by a Middle Line Similar to the lanes assigned peaks can be labeled using the Configure button of the Label pane on the Bands tab Image Overlays Petra2 adf E E X Lares Bands Label OK Ell Configure Font Arial Size po prti print Bi 7 B l I Mark by W Position Border Cancel Figure 2 8 Image Overlays Dialog Box Bands Tab As for the lanes use the Font pane of the dialog box to set the font options font font size font style Specify the position of label relative to the band marks left center right using the controls next to the font style selection buttons To insert marks for the peaks on the image
13. Finally you can assign bands within profiles that were not automatically detected 1 Activate the Assign function E 2 Click directly on the middle of the band Performing 1D TLC Evaluations 93 If the automatic lane and peak detection tool does not lead to satisfactory results determine the peaks using the controls of the Peak Determination panel Feak Determination position by center HS T sensitivity FWHM pixel Figure 3 18 Peak Determination Panel Ella 1 Set the appropriate option for marking the position of the band using the Peak Determination pull down menu position by center E position by maximum wv position by center Figure 3 19 Peak Determination Pull Down Menu 2 Activate the manual peak assignment tool 3 Click on the start of the peak in the image or in the profile window hold the mouse button and drag the second mark to the end of the peak You can change the peak region by dragging on one of the marks 4 When the peak region is properly defined click the right mouse button or press the Enter key to confirm To delete currently selected peaks e Click this button To search peaks automatically within the active profile s 1 Set the Sensitivity and FWHM parameters to the appropriate values All these parameters should be tested out for a particular image type and can then be used for all further images of this type The selected parameters can be saved under specifi
14. TLC plate Figure 3 4 Editing TLC Overlay 3 Press the Return key to confirm the changes Adjusting Origin Lines 1 Double click the origin line overlay Figure 3 5 Editing Origin Line 1 Performing 1D TLC Evaluations 85 Figure 3 6 Editing Origin Line 2 In most cases a horizontal line will be sufficient If the origin lines do not run parallel 1 Double click the origin line overlay again to enter additional vertexes 2 Choose the appropriate Line Model from the respective pull down menu to fit the overlay with the origin lines Line Model FTILIDIITETTIDITITITIIIIITIEIIIIIIIIE S Circular Parabolic Figure 3 7 Line Model Pull Down Menu Note The Circular and Parabolic line models are not available for line overlays with more than three vertexes 3 Repeat the action for adjusting the front line overlay Figure 3 6 Adjusting non parallel Origin Line You can use the various combo boxes to type in directly the position and dimensions of your plate and origin front line if you know them H If you want to change the run directions after the overlays have been set you have to delete the already set overlays first by clicking the Delete Overlay button and then redefine them 86 AIDA ID TLC User s Manual Now you have completed the setting of the TLC plate and the origin front line overlays You may proceed now with the usual 1D evaluation Select the Lane amp Peak Determination
15. To Edges expands the densitometer window beyond its narrow sides over the entire length or width of the image If no overlay is selected only the basic editing functions of the standard contextual menu described in the AIDA Image Analyzer Basic Concepts and Features User s Manual will be accessible Image Overlays You can specify the label type and font you want to apply to the overlays lanes and bands using the Image Overlays command of the Options menu Options View Window Hi Image Attributes Image Overlays Preferences Automatic Export Configuration Path Image Search Path Aubosaving Overlay Colors User Registration Figure 2 5 Options Menu Opening the Image Overlays Dialog Box After choosing Image Overlays from the Options menu the Image Overlays dialog box opens allowing you to specify the labeling and marking of lanes and bands separately using the Lanes and Bands tab respectively In addition you can specify marking and labeling for RF Fronts 16 AIDA ID TLC User s Manual 2 For Lanes depending on the already performed evaluations various labels can be selected by clicking the Configure button in the Label pane of the Lanes tab Image Overlays Petraz adf E AX Lanes Label Ell Configure Cancel Fant Arial Size hoo rtia print BIB STZ w Mark by Middle line v Border lines Figure 2 6 Image Overlays Dialog
16. any of the table rows the row s contextual menu opens allowing you to Select Columns for displaying or Delete Standards from the table Select Columns Delete Standards Figure 2 44 Quantity Calibration Window Row Contextual Menu Columns Selection X Show Columns Hide Columns Profile Peak Rel Deviation Integral Bkg GL Deviation Std Quantity Difference Recale Quantity cancel Figure 2 45 Selecting Columns to be Displayed 44 AIDA ID TLC User s Manual The following columns are available Table 2 3 Columns Profile Peak Name Integral Bkg signal unit Std Quantity quantity unit Recalc Quantity quantity unit Difference unit Rel Error 96 Deviation unit Rel Deviation 96 Columns Available in the Quantity Calibration Window Short Description The index and name of the profile The number of the peak in the profile The name for the peaks as given by the user The background corrected integral of the peak The quantity of the peak as entered for the calculation of the quantity standard curve A calculation of the peak quantity based on the current quantity calibration curve The difference without sign between the entered standard and the recalculated value The relative difference without sign of the entered standard and the recalculated value compared to the standard value 7 Difference Std Quantity The difference with sign betwe
17. list item und click OK Note This evaluation mode is only available if you purchased the AIDA 1D TLC module Note further that there are two possible ID TLC evaluation modes in AIDA 1D TLC one with GLP features indicated by the acronym GLP in parentheses and the other without those functionality The 1D Thin Layer Chromatography GLP evaluation mode is only available after establishing access control by applying the user registration see the Create User Registration section below 6 AIDA ID TLC User s Manual Create User Registration Access control to AIDA might be required when working under Good Laboratory Practice GLP conditions AIDA provides three different types of users a System Administrator Administrators and normal Users The System Administrator 1s a fixed account in AIDA which cannot be changed or deleted When no password is set for the System Administrator the user registration in AIDA is deactivated Administrators have the permission to create modify and delete users or administrators For GLP relevant modifications of the program settings general administrator rights are required in AIDA Users can modify their own password They have access to the normal functionality of AIDA When the user registration is activated you must enter your user name and password before you get access to the program AIDA Registration Login X Please enter user name and password User name Administrata
18. 13 Zooming X Axis 3 Release the mouse button The scaling will change accordingly Using 1D TLC 21 a raytest This procedure is also applicable in 3D display mode LITTTITICEEERESAS o DITTTUTTUTCHNENEENENNSSESS Lid Peak Type Rel Pos Integral Bkg Integra Bkg RF Value Std Qi Peak Type Rel Pos Integral Bkg Integra Bkg RF Value Std Qi pixel PSL PSL PSL nc pixel PSL PSL PSL nc 1 Profile 1 1 Profile 1 Peak Type Rel Pos Integral Bkg Integra Bkg RF Value Std Qi pixel PSL PSL PSL nc z 1 Profile 1 Figure 2 14 Zooming X Axis in 3D Display Mode To zoom out again click the right mouse button when the 4 symbol is displayed The maximal zoom out factor is defined using the Data Scaling command see below The same procedure can be applied to the y axis 22 AIDA 1D TLC User s Manual r seam To zoom into the Profiles window using the Zoom tool 1 Click the Zoom tool on the toolbar TREE Figure 2 15 Selecting Zoom Tool 2 Draw a rectangle around the area of the plot you want to see in more detail e o E L Intensity PSL Intensity PSL 0 100 200 300 400 500 600 700 800 0 100 200 300 400 500 600 700 800 Position pixel Position pixel SelectProfiles wi SelectProfiles K gt pE Peak Type Rel P
19. Box 1D Report Table Export C Comma C Semicolon 78 AIDA ID TLC User s Manual You can specify the Range of the exported rows Header only All Profiles or Selected Profiles only as well as the List Delimiter and Decimal Delimiter in the respective panes of the dialog box Selecting the System Defaults button applies the Windows default settings to List Delimiter and Decimal Delimiter 4 Complete the action by clicking OK The data are saved to a simple ASCII text file Exporting Quantity Calibration To export Quantity Calibration Data 1 On the File menu click the Export menu item and then choose Quantity Calibration from the submenu 2 In the Quantity Calibration Export dialog box that appears type a name for the export file and specify the data format Quantity Calibration Export 4g test adf Save in E Examples t File name Ag test cal Save as type ADA Calibration Cancel Windows Metafile Table A5CII File AIDA Calibration k Figure 2 93 Quantity Calibration Export Dialog Box To export the graph in the Quantity Calibration window e Choose Windows Metafile The graph is saved to a separate file in the Windows Metafile format To export the result table of the Quantity Calibration window 1 Select the Table Ascii File data format 2 Click Save to confirm the export Using ID TLC 79 Qe ee S Y raytest The Quantity Calibration Export Options dialog
20. DA ID TLC User s Manual With the Data Scaling command you can configure the graphical output The Minimum and Maximum value of the x Position and y axis Intensity can be selected together with a logarithmic representation for y axis Both axis can be set to Automatic scaling using the appropriate checkboxes The 3D View Point settings are the numerical representation of the change in 3D views as described above Data Scaling X Position mrm Minimum Maximum T i Automatic Cancel Intensity GL Minimu haxirurm 754 7 UU M Logarithmic v Automatic 3D View Point A Position Y Position oo B E Be X Figure 2 28 Data Scaling Dialog Box The Data Smoothing function is used to smoothen the curve Smooth Width Pools1 adf Ea Set smooth width to mm Cancel Figure 2 29 Smooth Width Dialog Box Using 1D TLC 35 The Grid command opens the Grid dialog box which allows you to lay grids over both axis that are an extension of either the big or the small ticks on the axes Vertical None Big Tics C Small Tics Harizantal Mone C Big Tics C Small Tics Cancel Help Figure 2 30 Grid Dialog Box The Profile Overlays command opens a dialog box that allows you to select various labels for the peaks together with their font font size font style and rotation angle The display of peak Area and or Brackets indicating the bands borders can be sel
21. Expand To Egdes Figure 2 3 Contextual Menu Lane Selected Band Selection If you have selected a single band the contextual menu that opens after right clicking the respective object provides you with the possible commands for this type of selection These include commands for editing and labeling profiles lanes as well as commands for editing peaks bands Note that if you select multiple bands the peak editing command of the contextual menu is turned off K Delete Profile x Duplicate Profile Edit Profile Delete Peaks Edit Peak Image Overlays Change Profile Name Set Profile Width Flip Profile Expand To Egdes Figure 2 4 Contextual Menu Band Selected The Delete Profiles and Delete Peaks commands are only executed for the selected lane s and selected peaks respectively Using 1D TLC 15 With the Duplicate Profile commands you can insert another profile identical to the currently selected one Edit Profile allows you to reshape the currently selected profile The currently selected peak area band can be re defined with the Edit Peak command The Image Overlays command opens the Image Overlays dialog box described below Change Profile Name is used to change the name of the currently selected lane Set Profile Width is used to adapt the width of the densitometer windows drawn around the lanes Flip Profiles changes the densitometer orientation upside down and Expand
22. IDA ID TLC User s Manual To delete a peak 1 Right click on the hatched area indicating the peak s domain 2 Choose Delete Peaks from the contextual menu T e a n E Wl Profiles Pools2 adf B x rog 600 500 400 k 300 o a 200 reate Peaks 100 10 Delete Profiles Intensity GL Delete Baselines Select Profiles 4 Display ar Data Smoothing Scales Grid Profile Overlays 4 Figure 2 20 Selecting peak and Opening Contextual Menu A dialog box opens prompting you to confirm the command Using 1D TLC 21 3 Click OK to delete the selected peak acl Profiles Pools2 adf E B x Delete Pools adf x Delete selected peaks Cancel Intensity GL Figure 2 21 Confirming Deletion of Peak Alternatively you can delete peaks by first selecting the peaks in the Image window and pressing the DEL key on your keyboard 28 AIDA ID TLC User s Manual Toolbar of the Profiles Window Select Profiles Back Button Smooth Width Button Forward Button Subtract Baseline Button Multiple Button Hide Baseline Button Tile Display Button Show Baseline Button 3D Display Button Normalize Button Figure 2 22 Profiles Window Toolbar Using the Select Profiles button you can select any profile by its name With the arrow buttons next to the Select Profiles button you can browse for
23. LC Image Example Here you can see the TLC plate overlay together with the origin and front line 83 The TLC plate overlay is used for the calculation of the Total value shown in the Profiles window If the 1D TLC Calibration toolbox is active letters are displayed referred to by dimension and position combo boxes of the toolbox You may change the location and size of the letters by choosing Image Overlays from the Options menu Here you may also choose if the TLC overlays shall be drawn at all To set the TLC plate and the origin front line follow the steps below 1 Select the 1D TLC Calibration toolbox 2 Select the run direction by choosing the appropriate entry from the Run Direction pull down menu Run Direction Top gt Bottom Right Lett J Left Right bottom Top mnn Top gt Bottom rmm Figure 3 2 Run Direction Pull Down Menu 3 Click the Define Sample button This will create an overlay for the TLC plate containing a second overlay for the origin and front lines By default the position of the TLC plate overlay s edges is congruent with the edges of the image Figure 3 3 Define Sample Button If your TLC plate is not identical to the whole image 1 Double click near an edge of the image This will activate the Editing mode for the TLC plate overlay 84 AIDA ID TLC User s Manual GY y 2 Move rotate and stretch this overlay until it is congruent with the
24. Manual Contextual Menu of the Quantity Calibration Window The contextual menu of the upper pane of the Quantity Calibration window contains two commands Scales and Grid With the Scales command you can configure the graphical output After clicking the Scales command the Data Scaling dialog box opens Here the Minimum and Maximum value of the x Concentration and y axis Intensity can be selected In addition you can turn on logarithmic representation and automatic scaling of either axis separately by clicking the appropriate option Logarithmic and Automatic below the Minimum and Maximum spin boxes Data Scaling i X iauantity hinirurm haxirurm DENN Logarithmic Automatic cancel Integral Bkg GL Minirmurn Logarithmic Automatic Figure 2 42 Data Scaling Dialog Box Quantity Calibration The Grid command opens the Grid dialog box which allows you to lay grids over both axis that are an extension of either the big or the small ticks on the axes SSS 05 Vertical Mone C Big Tics C Small Tics Harizantal Mone C Big Tics C Small Tics Cancel Help Figure 2 43 Grid Dialog Box The table in the lower part of the Quantity Calibration window provides its own contextual menu Using 1D TLC 43 The column headings are equipped with the standard contextual menus described in the Profiles Window section of this manual If you right click on
25. Menu of the Quantity Calibration Window 43 Audit Trail WAnIdOW ski oe eto ae e woah d dex e de dee iced 46 Additional Toolboxes in ID TLC 1 III 47 HO LEC Calibration 1 00 IDO 4 235 9n i i ato ut v RES Pee ee V i 47 R n Direction Panel 2c b rd eec obe ace de Cac Robe SU RO eiut 48 Sample Jetinttion Panel aedes dudum qw ve rar ee eee ws 49 TEC Plate Detinitron Patel 224 cns Rx eSI HE eMe ea 49 Ornon Line Panels 3a iru ERU Uu Ee cv ota UR ice ead 50 DtontLine Panel uas e E RREERRIPIXRREAERRARXXEROUu oe eee ee as tee 50 Lane amp Peak Determination Toolbox 0 0 0 0 0 ccc eee 50 Background Determination Section 0 0 0 eee eens 54 Peak Determination SeCLHOD ssr ea m V mor Fea au Fam x e CER dn 56 Act al Region Sections cse e ce UI E E Ree ea dee RCEEERR RE EY eed 57 Quantity Calibration Toolbox rinser ads Re TR XS RO eR RM S 57 Stalidard Pafie sats debe 4e EXERCERE E308 9 ibo GE dt Ed Senha be battu 58 Calibration Guive Pane 5a quo x teen VE o o AUR SRL RR MURUS aH d 60 iii iv GLP Supporting Tools Toolbox 0 0 eee es 61 yAdmanistrauton Panes o ace jp eben E Oe hand ua eee ean a ee 62 Pata El ratlSewimes Patieisus cad Etha d E eL ERA apetece 63 LanishoEvaluatton Pane o bsc rodar rae Is UE EAR CEA CU ge A 66 Automatic Export Pane 2 oe e o ede C E TRUE REESE 67 Data Ent LTD ae een ah enna eave oa Seok 67 Publishing Layout Toolbox in ID TLC 2 0 nes 69 Print O
26. Pos Select Abs Pos 1 std 1 4 R amp 5td Hide Abs Pos Settings of Abs Pos tsd B RStd Twa lined Header Select Columns Aadi std 1 8 RStd 4 UD tsat f sf std 34 4072 4 Figure 2 34 Contextual Menu of the Profiles Window s Result Table With the first two commands of the contextual menu you can sort the table of the Result Table by any value column in ascending order A to Z or zero to 9 or descending order Z to A or 9 to zero Just choose the appropriate menu item Sort Ascending by Sort Descending by The Select Unselect commands of the contextual menu are used to select deselect a column to export The column heading of selected columns are highlighted To hide a column choose Hide command from its contextual menu You can recover hidden columns using the Select Columns command see below 38 AIDA ID TLC User s Manual To specify the decimal digits for the value output in a column choose Settings of from the contextual menu and enter the value for the decimal digits in the dialog box displayed Column Settings 4bs Pos X Decimal digits i i E OK Cancel Figure 2 35 Setting Output Format for Columns Use the One lined Header Two lined Header command to reduce or expand the column heading by one line the second line indicates the units of measurement To select deselect the columns that should be displayed in the Result Table 1 Choose Select Col
27. af lane 3 changed fram 12 pixel ta 85 pixel Width af lane 4 changed fram 12 pixel ta 85 pixel Width af lane 5 changed fram 12 pixel ta 85 pixel Width of lane 6 changed from 12 pixel ta 85 pixel Width af lane 7 changed from 12 pixel ta 85 pixel Width af lane 8 changed fram 12 pixel ta 85 pixel Reason Figure 2 74 Audit Trail Confirmation Dialog Box The desired modification is blocked until the user confirms the action by entering the intention for the change explicitly in the Audit Trail Confirmation dialog box imi Audit Trail Confirm X Sunday April 18 2004 15 38 34 02 0 Width af lane 1 changed fram 12 pixel ta 85 pixel Width af lane 2 changed fram 12 pixel ta 85 pixel Width af lane 3 changed fram 12 pixel ta 85 pixel Width af lane 4 changed fram 12 pixel ta 85 pixel Width of lane FTSEYRTHRTITER 2 012 E Width of lane Width of lane width of lane Please give a reason For the changes gt Reason Figure 2 75 GLP Error Message Box Using 1D TLC 65 As for changes during evaluation you can also set recording options for changes afterwards Just click the Changes Afterwards button on the Data Trail Settings for pane The Audit Trail Configuration Changes Afterwards dialog box appears allowing you to select the actions to be watched by the GLP system Im Audit Trail Configuration Changes afterwards X Annotation Overlay Create Madify M Template Import M Auto Export Parame
28. al Bka Integral Bkg AF Value pixel PSL PSL PSL 1 Profile 1 1 Bko 384 siD4 aioa 0D0 005 2 1302 15909 1280 14528 Qr 3 1881 17832 1083 16808 014 2 224018680 S35 15755 018 5 O22 7 4 Figure 3 20 Selecting Profiles After all peaks are properly determined the quantity standards can be assigned 1 Set the method for the intensity evaluation in the Lane amp Peak Determination toolbox to Intensity Summation Optical Density should be selected for color stained bands Intensity Summation for radioactive fluorescent stains and chemiluminescence applications and Averaged Intensity can be used for fluorescent stains and chemiluminescence applications intensity summation NM intensity summation averaged intensity optical density Figure 3 21 Setting the Intensity Evaluation Method 96 AIDA ID TLC User s Manual It should be noted that integration over the entire densitometer window should be done in sufficiently large densitometer windows to accommodate the entire band Averaging can be performed best with densitometer windows that cover only the majority of the bands and omit empty space around them 2 Assign bands representing a known quantity as standards for calibration by selecting the Manual Assigment Tool from the Standard controls of the Quantity Calibration toolbox and click on the appropriate band in the imag
29. box appears allowing you to set further option for the export of the result table Quantity Calibration Export Mans i X Range MW Header Cancel i All Rows Selected Rows f AllColumns Selected Columns Decimal Delimiter f Point List Delimiter Tabulator C Comma System Defaults Figure 2 94 Quantity Calibration Export Options Dialog Box C Comma C Semicolon You can specify the Range of the exported rows Header only All Profiles or Selected Profiles only as well as the List Delimiter and Decimal Delimiter in the respective panes of the dialog box Selecting the System Defaults button applies the Windows default settings to List Delimiter and Decimal Delimiter 3 Complete the action by clicking the OK button The data are saved to a simple ASCII text file In addition the calibration data can be exported to an AIDA Calibration file that can be imported into another image Exporting Audit Trail To export Audit Trail data 1 On the File menu click the Export menu item and then choose Audit Trail from the submenu 2 In the Audit Trail Export dialog box that appears enter a name for the export file 3 Complete the export action by clicking the Save button The data from the Audit Trail window are exported to an ASCII file Audit Trail Ascii File txt 80 AIDA ID TLC User s Manual Importing Templates To import previously saved templates to another equivalent image
30. by detecting marking of the lanes and bands Then the quantities are normalized by setting quantity standards as applicable The result is a fully evaluated image where all the information can be collected in a comprehensive printout To comply to GPL standards AIDA 1D TLC provides record keeping features in addition to its 1D TLC features All changes to a previously performed evaluation can be recorded and are saved to a AIDA Lock File alf The system administrator can fully configure the behavior of the GLP supporting tools requiring for example to prompt the user to explicitly account for all modification of existing evaluations Starting 1D TLC To select the 1D TLC mode 1 Click the Evaluation button in the toolbar A dialog box appears offering you a list of available evaluation modes E Change Evaluation Mode E x Available Evaluation Modes 1D Densitometry 1D Densitometr 1D Evaluation 1D Evaluation GLP 1D Multi Labeling 1D Runlength Determination 1D Runlength Determination 10 Thin Laver Chromatography 20 Densitometry 20 Densitometr GLFP 20 Multi Labeling 20 Thin Layer Chromatography GLP Annotation Array Matrix Crystallagraphwy Material Science Cancel Microarray m Lil e Mose Qoob ee we a dera S6 o Fa Figure l 1 Changing Evaluation Mode The list entries reflect the installed AIDA application modules 2 Select the 1D Thin Layer Chromatography
31. c names using the Preferences option of the Options menu 94 AIDA ID TLC User s Manual For FWHM you should choose the approximate full width at half max of the peaks to find 2 Select this button to initiate the automatic search procedure To search for individual bands 1 Choose this button 2 Click on the band Bands that are not found with any set of parameters can be double clicked when this function is activated to obtain the retention factor for them Delete peaks of no interest by opening a rectangle over them in the selection mode and deleting the selected peaks by hitting the Del key on your keyboard or alternatively choosing Delete Peaks from the shortcut menu Delete the selected peaks or profiles Delete Profiles Cancel Check the Profiles window for peaks of interest Performing 1D TLC Evaluations 95 Assigning Quantitative Calibration Standards The profiles determined by the Lane amp Band Evaluation function can be visualized by activating the Profiles command of the View menu Any profile can be selected using the Select Profiles pop up menu or using the arrows under the Profiles window Wl Profiles Petra 2 adf E B x Y 200 7n 7 ul m I E c qi l l 100 Hh Wk n ji Ha Hd E 4 E h on y i NH AR RB i 7 NAL 1 i oi iN MR 0 100 200 300 400 500 FOO s Position pixel Select Profiles E Call Peak Type Rel Pos Integr
32. click the Export menu item and then choose Template from the submenu 2 In the Export Template dialog box that appears type a name for the export file and confirm the action by clicking Save Or 1 Click the Export Template button on the toolbar to open the Export Template dialog box Ep Figure 2 88 Main Toolbar Export Template Button 2 Type a name for the export file in the File Name text box and complete the export action by clicking Save Export Template Pools adf 2 x Save in X 1D Samples amp ee Fae File name pools2 atf Save as type alba Template Format L ancel Figure 2 89 Export Template Dialog Box 76 AIDA ID TLC User s Manual Exporting Profiles To export Profiles 1 On the File menu click the Export menu item and then choose Profiles from the submenu 2 In the Profiles Export dialog box that appears type a name for the export file and specify the data format To export the graph in the Profiles window e Choose Windows Metafile The graph is saved to a separate file in the Windows Metafile format To export the profile data of the Profiles window 1 Select the Profile Ascii File data format 2 Click Save to confirm the export The Profile Export dialog box appears allowing you to set further option for the export of the result table Profile Export Pools2 adf X Range Iv Header All Profiles Cancel C Selected Profiles Decimal Delimi
33. d corrected integral of the peak The percentage of the peak integral in comparison to the Sum or to the value in the row which is Set to 100 94 if defined The percentage of the peak integral reduced by the background intensity in comparison to the Sum or to the value in the row which is Set to 100 if defined signal unit and length unit units of measurement depending on the type of the imaging device used to obtain the image and the options set in the Image Attributes dialog box quantity unit unit of measurement specified by the user through quantity calibration Using 1D TLC 31 Table 2 1 Columns Available in the Result Table of the Profiles Window Column Short Description Std Quantity quantity unit The quantity of the peak as entered for the calculation of the standard curve Recalc Quantity quantity unit A calculation of the peak quantity based on the current calibration curve Recalc Quantity The percentage of the peak quantity in comparison to the Sum of all peak quantities or to the value in the row which is Set to 100 if defined RF Value Retention factor value signal unit and length unit units of measurement depending on the type of the imaging device used to obtain the image and the options set in the Image Attributes dialog box quantity unit unit of measurement specified by the user through quantity calibration The following additional rows can be found or creat
34. e or the peak in the profile Lk Quantity Calibration Standard Test Manual Assignment Tool Figure 3 22 Manual Assigment Tool Quantity Calibration Toolbox 3 Enter the value and the dimension in the Quantity Calibration dialog box that appears Set standard of 11 peaks LOK in 1 profile Profile 1 Cancel Standard Clear Figure 3 23 Quantity Calibration Dialog Box 4 After all standards are entered select the appropriate Calibration Curve type that fits the data and application 5 For most applications with properly assigned background it might be advisable to select the 0 0 Origin option so that the standard curve goes through the origin 6 Check the fit of the standard curve The r value gives a clue about the fit it should be very close to 1 for a good standard curve The calibration curve can be selected on the basis of theoretical considerations or simply to fit the observed situation Performing 1D TLC Evaluations 97 To evaluate the accuracy of the fit of the curve you can check the difference between the quantity standard value and the recalculated standard value 5 x Linear Regression y 1 621 x Correlation Coefficient r 1 0000 100 Integral Bkg a u 0 10 20 30 40 50 60 70 8D 90 100 110 120 Quantity ng Profile Peak a u Quantity Std ng Quantity Recalc ng Relative Error 1 Profile 1 Figure 3 24 Quantity Calibration Curve Example
35. each densitometer curve in a separate graphic Multiple shows all selected curves on top of each other in a single graphic and 3D shows all selected curves in a 3 dimensional fashion within the same graphic 70 AIDA ID TLC User s Manual To turn on printing of the result tables for the quantification and or evaluation of the migration data click to select the Table box Single Tables prints a separate table for each 1D densitometer All in one prints all results combined in a single table To Graphic prints the tables together with its associated densitometer graphic on a single page Print Options for Quantity Calibration The second additional button of the Publishing Layout toolbox is used to configure the layout setting for the Quantity Calibration window Protocol Print Fie e Print Preview Quantity Calibration Format Portrait C Landscape Print W Graphic WW Table Figure 2 683 Print Option for the Quantity Calibration Window As for the Profiles you can select the paper orientation Portrait or Landscape by clicking the appropriate radio button in the Format pane The options in the Print pane enable or disable the printing of the calibration curve Graphic and or the calibration table Table Using ID TLC 71 Print Options for Audit Trail The last button of the Publishing Layout toolbox is used to configure the layout setting for the Audit Trail window in the Protocol Print pane
36. ected To turn off the marking of the peaks on the graphics display click to deselect the Draw Peaks option Profile Overlays Petraz adf P ES x Region Label Ell Configure v v Draw peaks Mark peaks by Iv Area Brackets Figure 2 31 Profile Overlays Dialog Box Label Tab 36 AIDA ID TLC User s Manual oo g j e S AN H The location of the RF fronts can be drawn into the profile by clicking the Draw RF Fronts option on the 1D RF Fronts tab Further you can specify the font size of the labels by entering a value in the Label Size combo box Profile Overlays Petra2 adf r a x ql 40 RF Fronts E M Draw RF Fronts Cancel Label Size E E Pt Came Figure 2 32 Profile Overlays Dialog Box 1D RF Fronts Tab With the Change Profile Names command you can re name the densitometer windows and the resulting profiles according to your preferences The names will appear in all result tables and image menus Profile Name Pools1 adf X Set name of 4 profile to Cancel Figure 2 33 Profile Name Dialog Box Using ID TLC 37 Contextual Menu of the Profiles Window Result Table Each column in the Result Table has a contextual menu attached To open the contextual menu right click on the column heading gray area ET Es A Profile Peak Type Abs Pos mm Std Aunlength bn Recalc Runle Sort ascending by amp bs Pos Sort descending by Abs
37. ed exclusively in the Region Report Table 2 2 Additional Columns in the Region Report Sum Displays the scientifically correct sum for the selected column Ttl Total Sum over the entire profile Remainder Ttl Sum Numbers in italics indicate that the value was interpolated over the end of the defined standard curve and is therefore only representing a tendency not a result When a field is not relevant for a profile a dash is displayed The numbers can be formatted as outlined below You can select the number and type of columns you want to display using the contextual menu of any column in the window see below To change the column order 1 Click the heading of the column to be moved 2 Press the mouse button and drag to the desired position in the table 32 AIDA ID TLC User s Manual EE 3 Release the mouse button Wl Profiles Pools2 adf Wl Profiles Pools2 adf Wl proriles Poolsz adf select Profiles a Select Profiles iki select Profiles JE Frofile Protile E Type Protile Ty Peak E std 3 2 RStd 5 std 3 2 RStd T std 3 Fistd 2 Figure 2 25 Changing Column Order To change the column width 1 Position the mouse pointer over the border of the column whose width you want to change 2 Click and drag the border in the appropriate direction 3 Release the mouse button Wl rrorilesPoolsz ll El ProrilesPoolsz ll WI Profiles Pools2 select Profiles select Profiles
38. el The Front Line panel of the toolbox provides you with options to determine the position and Line Model of the front line Enter the appropriate values for the front Distance A G and Distance B H of the plate as well as the Line Model Polygonial Circular Parabolic Front Line Distance A G isell 759 Distance B H pixel 759 Line Modal Polygorial Figure 2 51 Front Line Panel The units of measurement can be changed via the Image Attributes option of the Options menu Lane amp Peak Determination Toolbox The controls of the Lane amp Peak Determination toolbox allow you to define lanes profiles and peaks on the image for signal intensity evaluation 50 AIDA ID TLC User s Manual To display the Lane amp Peak Determination toolbox e On the main menu click Evaluation and choose Lane amp Peak Determination Alternatively you can click the Lane amp Peak Determination button on the toolbar A Lane amp Peak Determination Options Direction arbitrary Profile Width BB boon org Lane Determination p a Expand to Edges Humber of Profiles A Background Determination Bka from Regions m Please set hkg regions to define the background Peak Determination position hy center m Sensitivity FWHM pixel Actual Region Configure la Ela Peak Height PSL Figure 2 52 Lane amp Peak Determination toolbox Using 1D TLC 51
39. ems to turn them on or off Select the number of copies you want to print E Print 1D Thin Layer Chromatography GLP ag test tif 1 d E X Printer Kyocera Mita F5 1010 KX at Megi OK Edges Left 2 0crn Top 2 0em cm Header Right 2 0cm Bottom 2 0cm v Parameter Parameter and Image Options W Image wholeimade I overlays Profile Curves portrait v grayscale W scales Profile Table automatic size M overexposed Quantity Calibration Iv Audit Trail Profle Options current selection landscape Use Colors Audit s SIUS Copies include positive trail configuration v include complete trail configuration i Figure 2 85 Print Dialog Box Using 1D TLC 73 raytest The margins of the printout can be selected in the Edges part of the dialog box The Header button displays the following dialog box Page Header and Footer x Header r Footnote pm M file path v author did ril pm v page number v last change time Cancel v current date v software version Figure 2 86 Page Header and Footer Dialog Box Here the Header and Footer of the report can be configured to contain various image and evaluation specific data The Parameter data overlays the Image Profile Curves Profile Table and Quantity Calibration information can be selected in the left part of the Print dialog box In the right part o
40. en the entered standard and the recalculated value Recalc Quantity Std Quantity The relative difference with sign of the entered standard and the recalculated value compared to the standard value 7 Deviation Std Quantity signal unit and length unit units of measurement depending on the type of the imaging device used to obtain the image and the options set in the Image Attributes dialog box quantity unit unit of measurement specified by the user through quantity calibration Using 1D TLC 45 Audit Trail Window In the Audit Trail window you can view all GLP relevant changes to the currently active evaluation as defined via the Data Trail Settings for controls of the GLP Supporting Tools toolbox including timestamp user id evaluation status and reasons for evaluation changes indicated by the current user f Audit Trail c Program Files raytest Aida Image Analyzer de m ml x 1 Friday March 2b 2004 12 34 3701 00 System Administrator Administrator al 10 Thin Layer Chromatography GLF evaluation started Figure 2 46 Audit Trail Window To open the Audit Trail window either choose the Audit Trail command from the View menu or alternatively click the View Audit Trail button in the Data Trailing pane of the GLP Supporting Tools toolbox 46 AIDA ID TLC User s Manual Additional Toolboxes in 1D TLC In addition to the standard AIDA toolboxes Display Control and Publishing Layou
41. ermination tools Direction arbitrary T Profile vWidth PTB wien E Lane Determination re 1 4 Expand ta Edges number of Frafiles FI Figure 3 13 Lane Determination Tools To search for lanes only select this button To search for lanes and peaks simultaneously select this button 4 Click to the upper left corner of your lane hold the mouse button and drag a rectangle to the lower right of the lane 5 Release the mouse button 90 AIDA ID TLC User s Manual Z y RENo atit a Higa as B mom mum F r Lip e i i r b Deas hay coe Lex wa TL Ww SU pum ZEE coy rum ae eats SERIEM MTS E E Figure 3 14 Searching for Lanes And Peaks Images where the automatic lane detection does not work can be assigned manually Figure 3 15 Lane Determination Tools 1 Activate the manual lane setting tool g Performing ID TLC Evaluations 91 E 2 Click in the middle of the lane at the start point and drag the densitometer to the end of the lane h PL 1 TEH E L emen i d ot Ma i Figure 3 16 Determining Lanes Manually 92 AIDA 1D TLC User s Manual Using this free densitometer function every double click on a position in the frame will create a new flexible point which can be moved independently This enables the creation of curved densitometer windows Figure 3 17 Creating Curved Densitometer Windows
42. eview option of the Publishing Layout toolbox allows you to preview all pages as configured in the Publishing Layout toolbox before they are actually printed Clicking the Preview button opens the Preview window directly since all layout setting are already made through the Print Protocol pane LE Aida 1D Thin Layer Chromatography Preview Petra2 adf E E 8 x File Edit Evaluation Options View Window Help Dips c Page 1 of 5 Page 2 of 5 1D Thin Layer Chromakgraphy Tuesday Apri 17 2001 12 43 04 PM 1D Thin Layer Chromakgraphy Tuesday April 17 2001 12 4304 PM Turaylez Hon v TLC mager Pe ra2 zd modHed Page tors Trays Manuals v TLC magez Pe ra2 zd modHed Page 2015 Protles daa t hi CALL A CT T d ML LR I LM ENIRE Ke inh ul N TENNE e a Aubor Sys lem Adminis kalor ulher Sys em Adminis kalor Lar Changed Tuesday Aprii 17 2001 12 4103 PM Alda Development Version 3 10 0238 L Changed Tuesday Aprii 17 2001 12 4103 PM Alda Development Version 3 10 0238 Figure 2 87 Preview Example Using 1D TLC 13 Export and Import In AIDA 1D Evaluation various tables and graphs can be exported to be imported in other documents or programs In addition the calibrations and templates can be exported to be applied to other images The various functions can be found in the Export and Import submenus of the File menu Exporting Templates To export your own evaluation templates 1 On the File menu
43. f the dialog box Parameter and Image Options you can select Whole Image or Window Content from the first drop down menu Landscape or Portrait format can be selected from the second drop down menu The printing of Overlays Grayscales Scales and Overexposed can be selected The printout can be automatically fitted to the page or the relative size of the image representation can be selected in 9 o In the Profile Options pane you can specify to print the Current Selection or all profiles as well as the paper orientation Landscape or Portrait for the printout Finally using the Audit Trail Options pane you can specify whether to include information about the Data Trail settings at all and if yes to which extent configuration information should be printed Include Positive Trail Configuration Include Complete Trail Configuration 74 AIDA ID TLC User s Manual as ES Preview As for printing in AIDA there are two ways of previewing pages before they are actually printed Using the Print Preview option of the File menu or Using the Preview button on the Publishing Layout toolbox The Print Preview option of the File menu opens the Preview dialog box which allows you to specify the layout settings for the preview The options presented here are the same as in the Print dialog box and those selectable from the various panes of the Publishing Layout toolbox However they are not stored with the data file The Pr
44. ged in as system Administrator to change the GLP settings If you are logged in as normal user the GLP Supporting Tools toolbox is considerably reduced providing you with the Data Trailing controls only njaj ajale fe GLP GLF Supporting Tools Data Trailing View Audit Trail Stop Evaluation Figure 2 69 GLP Supporting Tools Toolbox User Account Administration Pane In the Administration pane of the GLP Supporting Tools toolbox you can configure all GLP support settings Administration Iv apply GLP support proof PCA check sum Reason List Figure 2 70 Administration Pane Initial State Since this is only possible if no evaluation has been started ensure that no image is loaded into AIDA yet To enable GLP support Click the Apply GLP Support option The Proof PCB Checksum option enables the PCB image authentification feature Note Older PCB images do not incorporate checksums So if you want to apply the GLP Supporting Tools controls to those images make sure that this option is disabled 62 AIDA ID TLC User s Manual oo g j e S AN H The Reason List button opens the Audit Trail Reason List dialog box for predefining evaluation change reasons x Modity Delete Eu Figure 2 71 Audit Trail Reason List Dialog Box Data Trail Settings Pane The behavior of the GLP recording function 1s configured using the Data Trail Settings for controls Data Trail Sett
45. iles SelectProfiles 4 Eee hes ARAA Toolbar Peak Type Rel Pos Integral Bkg Integral Bkg RF value Std pixel PSL PSL PSL 1 Profile 1 1 Bra 34 sme soe Oo 005 2 1302 15909 1280 14528 Qr Result Table NENENECSELCHECENMEC CNET fed Ml TARA NI FAA 1h h h 113 4 k Figure 2 11 Profiles Window Example In the graphics pane of the Profiles window the displayed range of both axis can be zoomed To select a display range on an axis 1 Choose the Select tool from the toolbar Figure 2 12 Select Tool on the Main Toolbar 20 AIDA ID TLC User s Manual 2 Click at the smallest value of interest on the x axis and drag to the biggest value of interest on the same axis with the left mouse button pressed LITTTITOCEENENEEEESSS o LITTMTTCTUNEENENNSSSSM M Lind Intensity PSL Intensity P SL 0 100 200 300 400 500 600 700 S800 Position pixel Position pixel DBE A a DIES ie XT TC E Peak Type Rel Pos Integral Bkg etn ie RF Value Std al Peak Type Rel Pos Integral Bkg Integra Bkg RF Value Std Qi pisel PSL PSL pixel PSL PSL PSL nc 1 Profile 1 1 Profile 1 Man Intensity P SL 300 400 500 Position pixel MEEA A PEEM Peak Type Rel Pos Integral Bkg dis is RF Value Std pixel PSL PSL 1 Profile 1 b Figure 2
46. ing peaks to one peak after the region is confirmed After setting all necessary options click OK to return to the original toolbox content Densitometer windows can be drawn Arbitrary in any angle Horizontal Vertical or in Run Direction Select the appropriate direction from the Direction pull down menu The controls of the Profile Width section are used to automatically or manually determine the lane width Direction arbitrary Profile Width ok n pixel 8e Lane Determination p ae a i Expand to Edges Humber of Profiles Las Figure 2 55 Profile Profile Width Lane Determination Panel Select this button to enable automatic lane width measurement This button is used to determine the lane width manually I Clicking this button activates the manual lane setting tool Pi These two buttons activate the automatic lane detection tool with different options If the left button is selected only lanes will be searched Clicking the right button initiates the combined detection of lanes and peaks E3 Use this button to delete a selected profile If the Expand to Edges option is checked the lanes that are set by using the manual lane setting tool or the automatic lane detection tool will be expanded to the edges of the TLC plate overlay if one is set or to the edges of the image if no TLC plate overlay is set Using ID TLC 53 The miniaturized Profiles window on the far left side of the Lane Dete
47. ings for Evaluation Changes Afterwards Figure 2 72 Data Trail Settings Pane Using 1D TLC 63 1 Click the Evaluation button to set the recording options for changes during evaluation The Audit Trail Configuration Evaluation dialog box appears imi Audit Trail Configuration Evaluation E X Annotation Overlay Create Iw Modify M Delete Measurement M Template Import Auto Export Parameter Lane Create Iw Modify Iv Delete Lane Mame C Lane Number Lane Int Mode Feak Region Create Iw Modit M Delete Peak Name Background f Create Iv Modify M Delete TLe Calibration Sample Create Iv Modify M Delete Quantity Calibration v Std list M Equation Standard Create Iv Modify M Delete cael _ Figure 2 73 Audit Trail Configuration Evaluation 2 Specify which user actions should be recorded This includes for example the modification deletion of annotations changing calibration method and or calibration curve or modifying deleting regions 64 AIDA ID TLC User s Manual If the working operator performs one of the actions checked in the Audit Trail Configuration dialog box a dialog box appears prompting the user to give reasons for the desired change E Audit Trail Confirmation E x Sunday April 18 2004 15 38 34 02 00 Width af lane 1 changed from 12 pixel ta 85 pixel Width of lane 2 changed from 12 pixel to 85 pixel Width
48. ks To search peaks automatically within the active profile s this button should be clicked after choosing the appropriate Sensitivity and FWHM parameters Use this button to search for individual bands by clicking on them 56 AIDA ID TLC User s Manual Actual Region Section The Actual Region section provides information about the selected region Use the Configure button to add or delete categories from the list shown below the Configure button This opens the Column Selection dialog box described in the Profiles Window section of this manual Actual Region Configure Peak 10 Height FSL 166 5 RF value 0 70 integral 5 13 10 Figure 2 63 Actual Region Section Quantity Calibration Toolbox The controls of the Quantity Calibration toolbox allow you to perform quantity calibrations of 1D densitometers by assigning amount standards to the bands lE Quantity Calibration Standard user defined Standard pane Calibration Curve Logarithmic Regression Iv 0 03 origin Calibration Curve pane e Apo ng Corr Coeff rz 0 999882 Warning bkg not determined Figure 2 64 Quantity Calibration Toolbox Overview Using 1D TLC 57 To display the Quantity Calibration toolbox e On the main menu click Evaluation and then click Quantity Calibration Alternatively you can select the Quantity Calibration button on the toolbar Standard Pane L7 Quantity Calibration Standard S
49. ons for specifying the print options for the Profiles Quantity Calibration and Audit Trail Publishing Layout Annotate Z ai ale o Pratacal Print I Page Layout Edges Bottom 2 0m d Left 20m 4 Right 20cm 4 aN Header M File Path W Page Number M Current Date Profiles Options Button Quantity Calibration Options Button b Audit Trail Options Button IS Footnote jv Author M Last Change Time M Software Version Figure 2 81 Publishing Layout Toolbox Overview and Toolbar Using ID TLC 69 Print Options for Profiles Using the Profiles Options button you can configure layout setting for the Profiles window 1 Click the Profiles Options button The appearance of the Protocol Print pane changes Protocol Print Print Preview Profiles Format Portrait C Landscape Frint Selection Only Graphic Single C Multiple C 3p Iw Table C Single Tables Allin One Table to Graphic Figure 2 682 Print Options for the Profiles Window 2 Select the appropriate options In the Format pane select the paper orientation Portrait or Landscape by clicking the appropriate radio button In the Print pane the scope for the print output can be specified Click Selection Only to print only the densitometers that are currently selected Graphic selects the graphical representation of multiple selected profiles Single shows
50. os Integral Bkg Integra Bkg RF Value Std Qi Peak Type Rel Pos Integral Bkg Integral Bkg RF Value Std Qi pixel PSL PSL PSL nc pixel PSL PSL PSL nc 1 Profile 1 Intensity PSL Peak Type Rel Pos Integral Bkg neget 1a RF Value Std ee pixel PSL PSL a NH b Figure 2 16 Zooming into Profiles Window 3 Release the mouse button 4 Right click to zoom out Using 1D TLC 23 This procedure is also applicable in the 3D display mode with the difference that you have to draw the rectangle on the back plane of the cuboid 18 Apsua Position mm 128 Apsua Position mm ned N 18 Apsua Position mm Figure 2 17 Zooming into Profiles Window in 3D Display Mode AIDA ID TLC User 5 Manual 24 To change your viewpoint in 3D display mode Drag the axis origin with the mouse Intensity aL Intensity aL T a ir c D E Intensity aL E 3l 40 Position mm Figure 2 18 Changing 3D Viewpoint Using 1D TLC s 29 To edit peaks pk Double click the peak whose borders or position you want to edit Vertical delimiter bars appear nN Drag the vertical delimiter bar Intensity aL Intensity aL T a ir Cc D E Position mm Figure 2 19 Editing Peaks 3 Right click to confirm the changes 26 A
51. ptions Tor Profiles 2440 28 sa lt ewa gues GSS Ebene Oe eee ee tee See 70 Print Options for Quantity Calibration 0 00 ees 71 Print Options Tor Audit Trall 4 oem C s god A a T2 lor wc CP eT 73 IIEVIGWLS d db iare Eae ilius Cease gae eia ud apa Ea dad aod adis 75 Bxpotrtand PON 2123 idani kan e eae hue onde a E ti et A baee is 76 Exporting Template Sus vea d oc aeu sen EE AR dun E d RR ede ae RES ates 76 Expor PIOMles eod aod menora fece Xov uoce SA Bates was de Roi urs a Exporting 1D Report Tables iu nann ae dul eh we a4 2 OR ADS Vo he ea Xo 78 Exporting Quantity Calibration 0 0 0 0 0 eee nes 79 Expottins Audit ETIIT 2c e zoe doe Pot des DOM he Sdn HOSS ee hh UL des 80 Maporma Ternnplate Suse s coe aci rad vr Rp se bead hat ae eed ee 81 Importing Quantity Calibration 0 0 0 ee es 82 Performing 1D TLC Evaluations 83 Setting the TLC Plate and the Origin Front Line 0 0 0 83 Assigning Lanes and Bands 000 cc eee eee nee 88 Assigning Quantitative Calibration Standards 0 0 cece ees 96 Configuring the Evaluation Output 0 0 0 0 0 99 AIDA 1D TLC User Manual Introduction 1 Overview The AIDA 1D Thin Layer Chromatography module henceforth 1D TLC facilitates the analysis of 1D applications using quantification and retention factor RF value evaluation The first step in the evaluation is to set the TLC plate overlay and the origin and front line followed
52. rmination section displays the currently selected profiles of the densitometer window Lane Determination T Figure 2 56 Lane Determination Section The Number of Profiles is used only for the automatic lane detection tool If Fixed is selected the automatic lane detector assumes that the maximal number of profiles to be found is equal to the value selected from the combo box on the right of the Number of Profiles pull down menu Mumber of Profiles auto _h Background De TEST Bl a fram Benin Figure 2 57 Number of Profiles Pull Down Menu If Auto is selected the number is ignored and the automatic lane detector tries to find out the number of profiles itself Background Determination Section Perform background determination if you want to quantify bands after standardization There are two methods for determining the background the first one Bkg from Regions determines the background from regions set by the user in the Image or Profiles window the second one Bkg from Baseline uses a baseline set by the user in the Profiles window To set the base for the background determination choose one of the options provided by the Background Determination pull down menu Background Determination Bka From Baseline Please v Bkg From Regions Figure 2 58 Background Determination Pull Down Menu 54 AIDA ID TLC User s Manual Selecting Bkg from Regions allows you to set regions as background by clicking
53. s Quantity Calibration Audit Trail You can access these data windows by choosing the appropriate entry Profiles Quantity Calibration and Audit Trail respectively from the View menu M Window Help Image Histogram Profiles Quantity Calibration Audit Trail Figure 2 1 View Menu Image Window in 1D TLC All data are evaluated from the Image window Although it might not always be on top it is always open The title bar of the Image window shows the full file path of the image st Manuals 9 TLC Images Petrazadf_ Figure 2 2 Image Window Example You can copy the content of the Image window to the clipboard using the Copy Content command of the Edit menu See the Export Image section in the AIDA Image Analyzer Basic Concepts and Features User s Manual for further explanation 14 AIDA 1D TLC User s Manual Contextual Menu of the Image Window In 1D TLC the contextual menu of the Image window looks slightly different depending on the type of overlay you have chosen Lane Selection If you have selected one or more lanes the contextual menu that opens after right clicking the respective object provides you with the possible commands for this specific selection These include commands for editing and labeling profiles lanes Duplicate Profile Edit Profile Delete Peaks 4 Edit Peak Image Overlays Change Profile Mame Set Profile Width Flip Profile
54. s THIS PUBLICATION IS PROVIDED AS IS WITHOUT WARRANTY OF ANY KIND EITHER EXPRESS OR IMPLIED INCLUDING BUT NOT LIMITED TO THE IMPLIED WARRANTIES OF MERCHANTABILITY FITNESS FOR A PARTICULAR PURPOSE OR NON INFRINGEMENT THIS PUBLICATION COULD INCLUDE TECHNICAL INACCURACIES OR TYPOGRAPHICAL ERRORS CHANGES ARE PERIODICALLY ADDED TO THE INFORMATION HEREIN THESE CHANGES WILL BE INCORPORATED IN NEW EDITIONS OF THE PUBLICATION raytest MAY MAKEIMPROVEMENTS AND OR CHANGES IN THE PRODUCT S AND OR THE PROGRAMS S DESCRIBED IN THIS PUBLICATION AT ANY TIME Contents 1 Introduction 5 OV eWeek bee oie he E EE E E ep eed Bee EEEE EEA 5 Banlieue dee ebur du deese ha tite redire ta Oe Ge et 6 Create User Resistration cache aud varus tS needed ese Gt doa a E quede 7 2 Using 1D TLC 13 Data Windows AID DE ossiani ac ERE ae Rn a e OE red BS dee OS 13 Imasec Window ti IDIL Cs seht ad er at a eee envie hes penu hee Gee 14 Contextual Menu of the Image Window 000s 5 Mare Overlay sabe bas enun dex Seve dete m detached iere ut etd 16 Probes M INdO Wor ss See oS 28 h45 SRE PRATER SASS ADORE EASE e onn i s 20 Toolbar of the Profiles Window us ves ER CER GS 29 Data Descnpuon 4 3654462 EORR REQUE sone bie eee Gee ds 31 Contextual Menu of the Profiles Window Graphics Pane 34 Contextual Menu of the Profiles Window Result Table 38 Quantity Calibration Window 0 0 cence eens 42 Contextual
55. select the appropriate checkboxes in the Mark By part of the Image Overlays dialog box Position and or Border Besides the labeling for lanes and bands the Image Overlays dialog box allows you to configure the display of RF front marks and labels Just click the second icon on the toolbar to switch to 1D RF Fronts tab T Figure 2 9 Image Overlays Dialog Box Toolbar 18 AIDA ID TLC User s Manual oo P4 AX lt Z y LESS E To turn on the drawing of RF fronts on the image click to select the Draw RF Fronts option You can change the font size for the labels using the Label Size combo box In addition the Location of the Corner s Labels can be selected The Inner option includes the labels within the image whereas Outer puts the labels outside Image Overlays Petra2 adf X ol 1D RF Fronts 3m Iv Draw RF Fronts Label Size m Pt Location af Corners Labels Cancel di pes m k outer Figure 2 10 Image Overlays Dialog Box 1D RF Fronts Tab Using 1D TLC 19 Profiles Window The Profiles window is bipartite the upper pane displays the change of intensity over the length of the densitometer window integrated or averaged as selected over the width of the densitometer window the lower one contains a table listing the standards their given and recalculated values etc WI Profiles Petra2 adf B x Graphics Pane Intensity PSL Position piel Select Prof
56. stration User Management X Add Modify Password Delete ii User informations Full name System Administrator Permission Administrator Comment Fited account for the administration of the user database Figure 1 4 AIDA Registration User Managament Dialog Box 8 AIDA ID TLC User s Manual To add new users 1 On the AIDA Registration User Management dialog box click Add to open the Add User dialog box Add User x User name idoe Full name Jahn Dae Permissions iiser Password Em Confirm password Comment Cancel Help Figure 1 5 Adding Users 2 Type a user name full name and password for the new user and confirm the password by reentering it in the Confirm password field A User name can consist of up to 31 characters and must be unique in the registration database The Full name is used whenever the program has to display the name of the operator e g on print outs It cannot be longer than 63 characters The user s Password must contain at least 4 characters maximum 8 characters 3 Specify the user s permission level Currently two permission levels Administrator and User see above are available 4 Enter an optional comment for the account The Comment for a user account appears only within the user registration database 5 Click OK to apply the changes Introduction 9 To modify an existing user account
57. t the following toolboxes become available after selecting the 1D TLC mode TLC Calibration Lane amp Peak Determination Quantity Calibration e GLP Supporting Tool You can access these additional toolboxes from the Evaluation menu or by clicking the appropriate button on the toolbar cue Evaluation Options wiew Wine Display Control Publishing Layout TLE Calibration Lane amp Peak Determination Quantity Calibration w GLP Supporting Tools Figure 2 47 Mapping between Evaluation Menu Entries and Toolbar Buttons 1D TLC Calibration Toolbox The 1D TLC Calibration toolbox is used to set the TLC plate and the origin front overlays To display the 1D Calibration toolbox On the main menu click Evaluation and choose the 1D TLC Calibration entry Or e Click the 1D TLC Calibration button on the toolbar Using 1D TLC 47 eee ql 10 TLE Calibration Run Direction Right Left Run Direction Panel Sample Definition iu x Sample Definition Panel TLE Plate Definition A x pixel r2 F Y pixel T Dimensions TLC Plate Definition Panel A D pixel r52 A B pixel 5g2 Angle Origin Line Distance A E pixel 52 F Origin Line Panel Distance B F pixel 52 F Line Model Polygonial Front Line Distance A G pixel 700 E Distance B Hie 700 peepee Line Model Polygonial Run Direction Panel From the Run Direction pull do
58. tandard Selection Drop Down Menu ire st AutoAssign Tool Manual Assignment Tool ny 3 Delete Standard Assignment Button List of Standard Values Edit Standard Button Figure 2 65 Standard Pane Overview Manual Assignment Tool This button allows you to assign amount standards manually to the bands by entering appropriate values via a dialog box AutoAssign Tool Use this button to semi automatically apply Standard values to bands one by one using a predefined list The predefined list can be selected from the Standard Selection drop down menu The specified standard values are displayed in the List of Standard Values pane below the drop down menu Delete Standard Assignment Button Using this button you can delete selected standard assignment s 58 AIDA ID TLC User s Manual Edit Standard Button Use this button to enter and edit predefined standards Clicking this button opens the Quantity Standards dialog box DNA ladder DNA ladder Unit Precision Default Add Modify Delete OK Cancel Figure 2 66 Quantity Standards Dialog Box In the Quantity Standards dialog box the Standard name and Values are entered and the dimension of the amounts can be selected The Add Modify and Delete buttons are used to alter the list of standards All standards defined should be saved for future use This is also true for one time standards because evaluation get frequently re done which means that the s
59. tandard values are lost unless entered in the list Using 1D TLC 59 Calibration Curve Pane In the Calibration Curve pane of the Quantity Calibration toolbox the mathematical calculation of the standard curve can be selected and the result is displayed in the graph at the lower end of the toolbox Calibration Curve Logarithmic Regression Iv 0 0 origin Gi ng Corr Coeff f 0 999882 Warning bkg not determined Figure 2 67 Calibration Curve Pane The Calibration Curve drop down menu provides the following functions Linear Interpolation Linear Regression e Logarithmic Regression Log Weighted Regression Linear Interpolation calculates lines to join each data point Linear Regression optimizes a linear graph so that the deviation of the actual points from the curve is minimized Logarithmic Regression is used to determine the standard curve with logarithmic characteristics which is found in many saturating effects e g absorption Log Weighted Regression can be described as linear regression using a double logarithmic scaling The result is a better optimization of the curve fit for the smaller values instead of a better fit for the higher values Linear Regression Click to select the Origin option to force the curve through the origin The correlation coefficient Corr Coeff of the estimation is shown below the curve A value of 1 identifies a perfect correlation a value of 0 indicates f
60. ter Delete Lane M Create IY Modify Iv Delete M Lane Name M Lane Number Iv Lane Int Mode Peak Region Create Madify Delete Create Madify Delete Quantity Calibration Iv Std list W Equation Standard W Create M Peak Name Background W Create Modify Delete ZEN Figure 2 76 Audit Trail Configuration Changes Afterwards Dialog Box Ive Modify Iv Delete Finish Evaluation Pane The checkboxes of the Finish Evaluation by pane allow you to specify the user action which indicates the end of an evaluation Finish Evaluation by v first save v first clase Iv explicit stop Figure 2 77 Finish Evaluation Pane If you click First Save the settings of the Changes Afterwards dialog box become active after saving the evaluation the first time First Close activates the Changes Afterwards settings when you close the Image window during the initial evaluation Explicit Stop allows you to mark the end of the evaluation anytime you want using the Stop Evaluation button on the Data Trailing pane 66 AIDA ID TLC User s Manual Automatic Export Pane Automatic Export Edit Setting Figure 2 78 Automatic Export Pane The Edit Settings button of the Automatic Export pane allows you to select edit and save settings for automatically exporting the Profiles Quantity Calibration Runlength Calibration tables when the evaluation is saved to file Note If you are perfoming evaluations
61. ter f Point List Delimiter Tabulator C Comma Semicolon System Defaults Figure 2 90 Profile Export Dialog Box C Comma You can specify the Range of the exported rows Header only All Profiles or Selected Profiles only as well as the List Delimiter and Decimal Delimiter in the respective panes of the dialog box Selecting the System Defaults button applies the Windows default settings to List Delimiter and Decimal Delimiter 3 Complete the action by clicking Ok The data are saved to a simple ASCII text file Using ID TLC T Exporting 1D Report Table To export the 1D Report Table 1 On the File menu click the Export menu item and then choose 1D Report Table from the submenu 2 In the 1D Report Table Export dialog box that appears type a name for the export file 1D Report Table Export Ponls2 adf xl Save in E Examples amp rer E3 wem Fr Save as type Table ASCII File Cancel Figure 2 91 1D Report Table Export Dialog Box 3 Click Save to confirm the export The Profiles Export Options dialog box appears allowing you to set further option for the export of the result table Profiles Export Options k x Fange Ok M Header Cancel 6 All Rows Selected Rows Cancel AllColumns Selected Columns Decimal Delimiter C Point List Delimiter Tabulator f Comma System Detaults Figure 2 92 Profiles Export Options Dialog
62. ters should be set 1 First set the direction of the new lanes using the Direction pull down menu 2 Then determine the width of the lanes a Select the automatic lane width measurement tool 88 AIDA ID TLC User s Manual E b Click next to the upper left corner of a clearly visible band and drag to a position next to the lower right corner of the band Release the button Figure 3 10 Selecting Lane AIDA determines the width of the band and enters it into the appropriate combo box in the toolbox If no TLC plate overlay is set the automatic lane width measurement tool uses the direction selected via the Direction pull down menu If a TLC overlay is set the tool assumes the lane s direction to be parallel to the TLC overlay If the automatic lane width measurement does not lead to satisfactory results determine the lane width using the manual width measuring tool 1 Click this button then click the left side of a lane and drag to the right side of the lane Performing 1D TLC Evaluations 89 Sa me m om z Figure 3 11 Lane Width Value Indicated by the Lane Width Measurement Tool 2 Release the mouse button and press the Return key or the right mouse button If you know the width of your lanes e g from previous evaluations you should type it directly into the combo box next to the lane width measurement tool buttons Width Figure 3 12 Width Combo Box 3 Activate one of the automatic lane det
63. toolbox to assign lanes and bands and then use the Quantity Calibration toolbox to set quantity standards for normalization Performing 1D TLC Evaluations 87 Assigning Lanes and Bands The first step in the 1D evaluation is the detection marking of the lanes which is followed by the detection marking of the bands normalized by the setting of the quantity standards as applicable The result is a fully evaluated image where all the information can be collected in a comprehensive printout With AIDA 1D TLC it is usually preferable to select all parameters that are necessary for an automatic detection of lanes background and peaks first with one example lane When all parameters are well chosen further lanes can be automatically selected and assigned The detected bands can than be normalized by entering the values for the standard lanes quantification standards To begin with the evaluation of lanes it 1s important to set the Image Attributes in the Options menu first see AIDA Image Analyzer Basic Concepts and Features User s Guide If these settings are not correct further image evaluation will not result in meaningful results Image Attributes Petra pcb E E X Image Interpretatian Cancel Fired interpretation defined by image format Length Unit pixel Figure 3 9 Image Attributes Dialog Box When you use the Lane amp Peak Determination toolbox for an application the first time various parame
64. ull independence of values Good standard curves should have value very near to 1 60 AIDA ID TLC User s Manual Double clicking on the graphics pane opens the Quantity Calibration window see the Quantity Calibration Window section of this manual for details Sometimes a warning will be displayed below the graphics pane stating bkg not dertermined to indicate that no background region has been assigned yet It can also indicate the lack of a local background for some regions GLP Supporting Tools Toolbox The GLP Supporting Tools toolbox is used to specify all GLP relevant settings GLP GLF Supporting Tools Administration Iv apply GLP support proof PCA check sum Reason List Data Trail Settings for Administration Pane Evaluation Changes Afterwards Finish Evaluation by v first save v first clase Iv explicit stop Data Trail Settings Pane Finish Evaluation Pane Automatic Export Edit Setting Data Trailing Automatic Export Pane view orit Trail Data Trailing Pane stop Evaluation Figure 2 68 GLP Supporting Tools Toolbox Overview To display the GLP Supporting Tools toolbox e On the main menu click Evaluation and then click GLP Supporting Tools Or e Select the GLP Supporting Tools button on the toolbar Using 1D TLC 61 Note To configure the GLP support administrator rights are required Therefore make sure that you are log
65. umns A dialog box appears displaying two lists Show Columns Hide Columns 2 Select the columns you want to show or hide and click the appropriate arrow symbol to transfer the items between the two lists 3 To deselect a particular item just click 1t again Columns 5election E x Width mrm Recalc Quantity 95 Integral Bkg t Show Columns Start Pas mm End Pos mm cancel Figure 2 36 Columns Selection Dialog Box Using 1D TLC 39 In addition each row of the table has a contextual menu attached which includes commands for displaying special row types like Sum Total Remainder as well as for changing profile names iol EXITUS 3 i Frofile Peak Type Abs Pos mm Std Hunlenath bp Hecalc Rune 1 std 1 2AT faut aa 23 7 tadi 3 rsd 250 10120 tadi 4 RStd 28a A NENNT E Set to 100 2b 5 0 Bkg Region 8 6108 i4 Select Percent Reference i x 2038 Change Profile Mames 5 m kan mmw mi we Si icd 1 5 7 Select Columns Select Rows 1 1 M DE Figure 2 37 Contextual Menu of a Result Table Row Set to 100 will set the value in the active row to 100 instead of using the Sum value as 100 and recalculate the other values accordingly To select the columns that should be displayed in the Result Table choose the Select Columns command see above 40 AIDA ID TLC User s Manual
66. ward or backwards through the available profiles The three buttons in the middle provides you with different display possibilities for the graphical output Multiple displays multiple curves for selected items in a single two dimensional co ordinate system using different colors for each 3D shows all curves for selected items in a single three dimensional co ordinate system Tile uses a separate 2D co ordinate system for each curve of a selected item The Normalize button is used to normalize all displayed profiles according to the determined migration distance The Show Baseline Hide Baseline Subtract Baseline buttons are used to display the determined baseline hide the baseline and subtract everything below the baseline respectively This function is only available if a baseline has been set in the Band amp Peak Determination toolbox The Smooth Width button opens a dialog box to enter the smooth width for the curve display A higher number will result in averaging over the given range of pixel mm inch where the unit is set using the Image Attributes commands of the Options menu Smooth Width Pools adf ts X Smooth width 0 211 138 mm EI Figure 2 23 Smooth Width Dialog Box The lower part of the Profiles window contains a table henceforth Result Table listing the standards their given and recalculated values etc Using ID TLC 29 You can change the display ratio between the graphics and the table pane b
67. with GLP support disabled you can access the automatic export settings via the Automatic Export command of the Options menu Data Trailing Pane After starting an evaluation the controls of the Data Trailing pane are enabled Data Trailing View Audit Trail Stop Evaluation FCB check sum not proofed Evaluation is in process Finish evaluation by first save close the image explicit stop Figure 2 79 Data Trailing Pane Evaluation in Progress The View Audit Trail button is used to open the Audit Trail window Use the Stop Evaluation button to mark the end of an evaluation anytime you want In the lower part of the Data Trailing pane the status of the evaluation as well as the settings for ending the evaluation depending on the checked Finish Evaluation by options are given In addition if the PCB check sum option is disabled a warning is displayed below the Stop Evaluation button Using 1D TLC 67 After stopping the evaluation the action which marked the evaluation s end is displayed in this example by explicit stop via clicking the Stop Evaluation button Data Trailing View Audit Trail ker Evaluation FCB check sum not proofed Evaluation is finished by explicit stop Figure 2 680 Data Trailing Pane Evaluation Aborted by User 68 AIDA ID TLC User s Manual Publishing Layout Toolbox in 1D TLC In AIDA 1D TLC the standard AIDA Publishing Layout toolbox is expanded by four additional butt
68. wn menu you can select the run directions used to develop the TLC plate from which the image was obtained There are two vertical Top gt Bottom and Bottom gt Top and two horizontal options Right gt Left and Left gt Right available 48 AIDA ID TLC User s Manual Sample Definition Panel Use the two buttons of the Sample Defintion panel to create or delete the TLC plate overlay sample Definition Figure 2 48 Sample Definition Panel TLC Plate Definition Panel The various combo boxes of the TLC Plate Definition panel allow you to set the position of the TLC plate overlay by entering the appropriate values for the Position of Corner A the upper right corner of the plate and the horizontal Distance A B and vertical Dimensions Distance A D of the plate together with its Angle TLE Plate Definition Pasition of corner A X pixel Nj iu8 amp 8 z Y pixel Dimensions Distance A B fixe 968 amp Distance A D pixe 916 amp Angle 1 270 Figure 2 49 TLC Plate Definition Panel Using 1D TLC 49 Origin Line Panel The Origin Line panel of the toolbox provides you with options to determine the position and Line Model of the origin line Enter the appropriate values for the origin Distance A E and Distance B F of the plate as well as the Line Model Polygonial Circular Parabolic Origin Line Line Model Polygorial Figure 2 50 Origin Line Panel Front Line Pan
69. y dragging the delimiter line located on the bottom of the toolbar EA Profile Peak Tine Abs Pos mm Std A Intensity GL 0 0 20 30 40 50 F Position mm Select Profiles EE Figure 2 24 Changing the Display Ratio Between Graphics Pane And Result Table 30 AIDA ID TLC User s Manual Data Description Available Columns in the Result Table Table 2 1 Columns Available in the Result Table of the Profiles Window Column Profile Peak Type Name Rel Pos length unit Abs Pos length unit Start Pos length unit End Pos length unit Width length unit Height signal unit Integral signal unit Bkg signal unit Bkg 76 Integral Bkg signal unit Integral 9 6 Integral Bkg 96 Short Description The index and name of the profile The number of the peak The type of the peaks e g QStd for Quantity Standards The name of the peak as given by the user The position of the peak in the profile The position of the band in the image measured from the edges of the image The start position of the peak in the profile The end position of the peak in the profile The width of the peak The maximum height of the peak The integrated peak intensity The integrated background intensity The percentage of the peak background in comparison to the Sum of all peak backgrounds or to the value in the row which is Set to 100 if defined The backgroun

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