INFINICYT , 1.1 VERSION
Contents
1. 4 User can configure the mathematical transformation of the parameter SSC to improve the separation of populations cece cece cc cece eee eee e cece eae neeee scenes New options regarding printed report ccc cece cece eee e eee e eee e ee eee eee e ees 5 New options regarding Merger Module 5 1 5 2 New information and commands in the merger screen ccc cece seen eee e ee eens News regarding merger process controls ccccccc cece eee e eee e cece cece ee eeneenees 6 New opion tecardine FNGCEXDO nianon ileal aia oe aa es 10 11 13 14 14 15 15 17 17 19 1 The FL based merger option has been deleted In the version 1 1 there is a unique merger option since it is possible to obtain the information that in the previous version was obtained with the FL based merger option from the new Parameter Band Dot Plot diagram 1 Select the population of interest in the file resulting from the merger o a x w I o g I o 1 2 TCRgd FITC FL1 H Log 2 Open a new diagram type Parameter Band Dot Plot selecting as number of dimensions the number of parameters that you want to visualize from the merged file NEW DIAGRAM Histogra Qo aA Dotols BoE U ES Band DotP lot APS Graph Density 3D DatPlot MultiDimensional C Humber of Dimensions 3 By right clicking in the middle of the n2. the user can modify the compensation values manually so that the changes will be applied to the file In addition the user can save the values of the new compensation and apply them to other files These options are available in the Current Compensation Window of the Profile Configuration menu amp Profile Configuration a Preferences Analysis Graphs H Controls H Statistics H Stored Compensations i Gates 9 ammm s R Visibilty Configure APS Parameters H Properties Compensation data in Column Row A tose 549 pa 35 31 O S 0 78 aa i 00 I 00 E 34 ei O07 0 01 0 21 Cues a i 12 0 00 0 38 Mra 09 6 74 FA 789 131 0 29 ae 0 00 4 4 xrIno7 monorp FITC A CD22 PE A CD23 PerCP Cy5 5 A4 CD19 PE Cy7 amp 4PC A CD20 APC Cy7 A Pacific Blue A AmCyan A Selecting Stored Compensations option of the Profile Configuration menu the user can check delete and change the current file compensation for one of the compensation already stored Profile Configuration X Preferences H Analysis H Graphs H Controls H Statistics Stored Compensations Gates mars File Current Compensation Negative Visibility Configure APS Parameters Properties Stored Compensations 1 Draw consecutive gates to identify a population A B C D E F G H PerCP Cy5 5 4 PE Cy 4 APC
3. independently This option is available from the Configure APS Parameter window of the Profile Configuration menu and it is especially useful in merged files where we will select only the common parameters for the APS calculation W Profile Configuration gt Preferences Configure APS Pa Analysis Graphs Contrals Statistics F5C A Stored Compensations Gates Sunan Current Compensation oo Negative Visibility Configure APS Parameters v PercP cy Properties C PE Cy A rameters Select all parameters LINEAR Exp 55C Low LJ FCS file FITC 4 CD22 LOGICAL PE A CD23 LOGICAL s o ACD1S LOGICAL LOGICAL APC Cy 4 Pacific Blue 4 Ameyan 4 The aim of APS graph is for the user to be able to distinguish in these representations as many different populations as possible especially useful when working with a high number of parameters for a further characterization of these populations in the conventional 2D graphics New in this version whenever you gate and classify a population the APS graph will calculate again the view taking into account selected populations This way it 1s possible to distinguish even more subpopulations among already classified events GATE APS Joe granulocytes APS VIEW 1 GATE APS Jog EATA 4 N eosinophils EOM ait os ages n 2 neutrophils APS
4. FILE 5 b In order to avoid errors in analysis the option of correcting the parameters when a difference of no coherence between populations Population Control is detected has been deleted _18 6 New options regarding File Export 6 1 The available export file formats have been reduced to Standard FCS or TXT When FCS format is chosen the data will be saved in the form of an FCS file readable by any cytometric analysis software but not by common worksheet or database programs When it is chosen TXT format the values will be saved in ASCII format as a data base readable by worksheets or word processing programs 6 2 Some options on the advanced functions of the File export to avoid extra memory requirements resulting from unnecessary information have been added The user can export only the parameters which are necessary for the analysis Initially all the parameters are selected for export If the user wants to eliminate some of these parameters unclick the parameter to eliminate it from the export Events if the user is trying to export a merged file Infinicyt will give the option to either export events proportional to each file maintaining the file number separation or mixed in a file without separation between original files T FILE EXPORT Byteord 4 3 2 1 Datatype Restore Values Pn format Sudca Parameter PnA FS C Height FSC H iSSC Height SSC H a os ICD25 PE FL2 H I
5. matrix with the information for all the parameters and tubes Profile Configuration C Preferences Analysis Graphs Controls Statistics Stored Compensations Gates C J FCS file Curent Compensation Negstive Visibility Configure APS Parameters Properties Parameters Fie SSCA CD22FITCA CD13 PerCP Cy5 5 4 PEGA CD20 4PC A F5CA FMC7 FITC A CD24 PEA CD34 APCA CDIO3FITCA CO25 PE A CDIIC APC A amp Profile Configuration C Preferences Parameters Info oa Analysis Graphs Controls Statistics Stored Compensations Gates Parameters a Current Compensation c Negative Visibility u Configure 4P5 Parameters i Properties Pacific Blue 4 Am yan lt mT _ 3 4 New options make the Manual Analysis of the data file easier 4 1 The way in which the user can save and find in his computer a document already analyzed cyt has been improved Infinicyt software allows saving the files analyzed containing the information of classification statistics graphics report profile etc and store them in the hard disk Select Save Analyzed File option of the File menu when you want to store the analysis of a file The software will ask you the directory where you want to save it and the file will be saved as cyt ANALYSIS Normal peripheral blood sample Cctrl O Open Next File Ctrl N ulation Open Open Previous File Ctrl P Expo
6. 4 APC Cy 7 4 Pacific Blue 4 AmCyan 4 When the user selects a compensation already stored and clicks OK then the selected compensation will be applied to the file but whenever a new file is opened it will be always opened with its internal compensation 3 3 New option for creating consecutive gates to identify a population and save the sequence to apply it to similar files 10 amp E amp 200 400 700 1000 200 400 700 1000 z z z o 5 d al 5 r D A o a f a Wi W hr E E g E 5 f e 0 br4 we v rA o a rr Rott 0 1 FL2 H CD34 PE Loy FL1 H CD45 FITC Log FSC H FSC Height Lin 2 Go to Selection menu of the Analysis Window and select Save Gate As Help Windows File Statistics Paama Profile Undo Draw Region Ctrit 2 Redo Reset Analysis Chri R Delete All Selections Ckrl 0 Select Ei 60000 5116 853 853 54884 947 APE Pe 0 ooo 000 Edit Gates a00 0 50 0 55 ae 27771 4620 OST bo 2655 ARE A BAY 24008 40358 AA 0 ooo 000 27137 352z APTN i 955 159 3 716 353 BMZ k Save Gates 45 k K s x k k s Gate Name CD34 CELLS 4 When you want to apply the same gating sequence to a new file go to the Selection menu of the Analysis Window and click Load Gates Choose the appropriate gate from the list and then go to the Analy
7. CD19 PERCPCY5 5 FL3 H ICD11C APC FL4 H FMC FITC FL1 H CD24 PE FL2 H CD34 APC FL4 H CD43 FITC FL1 H ICD79B PE FL2 H Saas ES 3 53 KS KS KS 6 ES ES KS KS Events 250000 Proportional to each File Proportional to each File Mixed in a file 6 3 From the Main Screen the File Export option will consider the export of a whole file If the user is interested on exporting only one or more populations then the Export option must be opened from the File menu of the Analysis Window If the box relating to Only visible populations is enabled only the visible populations in the Analysis Window will be considered for exporting If it is not all the events contained in the file will be exported FILE EXPORT Basic Advanced Only visible populations Simulated Time Export File Format Standard FCS TXT 19
8. CP Cy5 5 A CD19 H Properties PE Cy 4 APC A CD20 APC Cy A Pacific Blue 4 AmCyan A FSC A Apply to all If we increased the negative visibility percentage applied for a certain parameter the negative part of the scale of this parameter will be less represented although it continues being visible CD19 PerCP Cy5 5 4 Parameter Yo FSC A 0 01 os CD56 PE Cy7 A 2 92 E a CD3 APC A 4 2 CD20 APC Cy7 A 3 p i CD45 Pacific Blue A 2 9 g z T gt 1E2 1E3 CD3 APC A LOGICAL Parameter oF FSC A b di a S5C A 0 q1 CD7 FITC A Ddi i CD4 PE A nder br CD19 PerCP Cy5 5 A iolss F m CD56 PE Cy7 A 2 92 CD3 APC A C 3 99 a CD20 4PC Cy7 A B 6 a CD45 Pacific Blue A 2 09 ha rr 7 10000 1000 CDJZAPC A LOGICAL e 3 5 New option to check all the parameters measured of a data file Properties It is possible to go to the information about the parameters measured on a given data file by clicking Properties of the Analysis Window File menu by clicking the Properties option of the Profile Configuration menu or with Crtl I This is especially useful when working with files acquired with more than 3 4 fluorescences or with merged data files because in this window we will always have available the
9. T TUBE 04 Configure Info Parameter 1 j2 3 4 H Add Select Files C Fsc a Cassa COJCD22 FITC A FITC A C cb23 PE 4 PE A mori C CD19 PercP Cy5 5 4 PerCP Cy5 5 4 CO JCD20 APC A APC A C CD103 FITC A FITC A C CD25 PE 4 PE A C C10 APC A APC A LO FMc FITE A FITC A C CD24 PE A PE A LG icb34 APC A APC A Save As Main Meng serra C KAPPA PEA PE A E Version 1 1 of Infinicyt includes the following available commands in the Merger Screen Configure File merge options allow manipulating the controls that will warn you in case your common parameters are not similar enough to perform a merge process Select files this command opens the directory where the user can select the files to be merged Once the data files are selected in the upper part of the dialogue box appears the list of the files to merge and in the lower part of the dialogue box appears the matrix parameters file o You can select and move the files on this list with the A and V cursors in order to obtain the best sequence to represent the tubes in the graphics o Inthe matrix the user can add or not each parameter in order to avoid parameters that are not interesting because there is no marker on it With this we will optimize the memory of the new data file reducing the number of parameters to be merged Save as when the selection of the data files is done and the parameters to merge a
10. To improve the benefits that this software offers Infinicyt 1 1 has included the following INFINICYT 1 1 VERSION 1 The FL based merger option has been deleted now there is only a type of merger 2 New options and types of Graphs have been created to show information 3 Itis possible to create and save Analysis Profiles for successive USES 0000 e ee 3 1 a2 3 3 3 4 a Statistics 3 1 1 Itis possible to use formulas to calculate results 0 eee 3 1 2 Absolute counts from the population of reference selected by the user Current compensation can be changed manually and saved if the origin FCS file contains this information internally cc ccc cece cece e cece eee e cece eeeeaneeenes New option of creating consecutive gates to identify a population and save the SEQUENCE LO apply IL 10 Siar TICS 22 oscuro hati aaseusddetiae ths eaeeoamnataenGer sae naaeaeanaads It is allowed to configure negative visibility of the parameters with Logical scale New option to check all the parameters measured of a data file Properties 4 New options to facilitate the Manual Analysis of the data file 4 1 4 2 4 3 4 4 It has been improved the way in which the user can save and find in his computer a document already analyzed CYL ss29s vessacrmsanerses oe eras ETO TRTE EE Option to choose the columns to visualize in the Analysis Window
11. VIEW 1 3 Itis possible to create and save Analysis Profiles for further uses In order to facilitate and standardize the analysis of similar samples version 1 1 of Infinicyt allows to configure to save and to load for further uses different analysis profiles ANALYSIS Increased CD4 cells PB sample File Statistics Selection Maian Windows He Configure Chril 6 Pop Load Profile Save Profile Partial BOOOO 100 004 100 004 0 0 00 0 00 0 0 002 0 00 0 0 002 0 00 we Gate 0 events Total 0 00 Visibility 0 00 Profile Configuration A profile As all the information about the software configuration It is possible to create different profiles P Graphs for different diseases so the user can always work with Lf Controls sas l Statistics the same graphs population tree controls statistics eee compensations gates report etc with the same type of di Gates Fes file study d Current Compensation It is recommended to see detailed information about pn Negative A Configure AFPS Parameters Analysis Profiles in chapter 5 1 4 of the User s Manual Properties 3 1 Statistics 3 1 1 Itis possible to use results calculation formula The result of the formula can be located in the populations tree if the created formula relates different previously established populations for example CD4 CD8 ratio or in the columns relating to the statistics in the Analysis W
12. ault Exp S5C Low Configurable Exp 55C Low Configurable Exp 550 Low Law 228 4 220 Helght Cag 35e a ma mae T pore Li Iria Wie E uea k 479 4 4 New options regarding the Printed Report 4 4 1 Itis allowed to copy cut and paste on Printed Report components of the software and external images or texts 4 4 2 Itis possible to choose the size of paper from the Properties option of the report File menu 4 4 3 Undo Cntrl Z Redo Cntrl Y options have been added from the Report Edit menu 4 4 4 Data option has been added as component of the printed report Add Component Choose a Component 4 4 5 Different options have been added to configure the populations the parameters and the statistics to be shown on the population component of the report In the version _15 1 1 of Infinicyt it is possible to select more statistic data choose different files and show comments if this option is selected in the columns menu Population AlE lect only populations with events O O O O O O O aa Rl C Columms SD Cy Median Mode New function a Display Options Indent Show Background 16 5 New options regarding the Merger Module 5 1 New information and commands in the Merger Screen FILE MERGE x 4 C001 PATIENT TUBE 01 2 C002 PATIENT X TUBE 02 j3 C003 PATIENT TUBE 03 j4 C004 PATIEN
13. ents Add Function Name CD4 CD8 Ratio CD4 T lymphocytes Partial 1 Comments Add Function Type Number Data Subfunction Population Data Partial Parameter Data Population CD8 T lymphocytes Ry C Other File Selected Comments Once the formula is finished click OK and the new function will not be a new column with values for each population but a unique value shown as a new population of the Population Tree ANALYSIS Normal peripheral blood sample File Statistics Selection Profile Windows Help Population Partial Visibility Comments Of aK 60000 6529 10 88 10 88 0 00 53471 89 12 89 12 100 00 0 0 00 0 00 0 00 1838 3 06 3 44 3 44 31582 52 64 59 06 59 06 3074 5 12 5 75 5 75 16977 28 30 31 75 31 75 Bove 0 0 00 0 00 0 00 irr 12438 20 73 73 26 23 26 f CD4 CD8 Ratio 0 60 aa Ea ea Other T lymphocytes 584 0 97 4 70 1 09 CD4 T lymphocytes 4432 7 39 35 63 8 29 o i 7422 12 37 59 67 13 88 SB B Lymphocytes 2586 431 15 23 484 Other B Lymphocytes 0 0 00 0 00 0 00 a kappa B lymphocytes 989 1 65 38 24 1 85 Lambda B lymphocytes 2 66 61 76 299 ok az nso sesx M Gate 0 events Total 0 00 Visibility 0 00 K K K K K K K K 3 1 1 b In order to show the result of the formula in the columns relating to the statistics in the Analysis Window go to Pr
14. ew Parameter Band Dot Plot go to Graph Configuration menu and select Axes Configuration Through this command the user may select the parameters to visualize on each band of the new diagram Seles Axes Configuration x IP10 PE ACTIVADO FL2 H Log Band 1 IL 2 PE ACTIVADO FL2 H LOGARITHMIC Band IL 4 PE ACTIVADO FLA H LOGARITHMIC Band 3 IL 10 PE ACTIVADO FL2H LOGARITHMIC Band 4 IL 13 PE ACTIVADO FL2H LOGARITHMIC Band 5 TNFa PE ACTIVADO FL2H LOGARITHMIC Band 6 TNFb PE ACTIVADO FL2 H Band 7 glIFN PE ACTIVADO FL2 H LOGARITHMIC Band 8 eee LOGARITHMIC PUAA RT E H AAE oo glFN PE ACTIVADO FL2 H Log THES PE ACTIVADO FL2 H Log ees AT NF a PE ACTIVADO FL2 H Log eo il 13 PE ACTIVADO FL2 H Log IL 10 PE ACTIVADO FL2 H Log IL 4 PE ACTIVADO FL2 H Log IL 10 PE ACTIVMADO FL2 H IL 2 PE ACTIVADO FL2 H Log eee Parameter Band Dot Plot shows the legend of the analyzed parameters Each band represents the real expression of the parameter selected in the original file See detailed information about the different options related to the file merger process in chapter 4 2 of the User s Manual 2 New options and types of Graphs have been created to show information See detailed information about types of graphic representations in chapter 5 3 of the User s Manual In the version 1 1 of Infinicyt Parameter Band Histogram and Parameter Ba
15. indow 3 1 1 a In order to place the result of the formula as a new population in the Population Tree right click on the first label of Events of the Population Tree and select Add Function ANALYSIS Normal peripheral blood sample SEE File Statistics Selection Profile Windows Help Population Events Total Partial Visibility Comments lt Of aK e0000 Dther Events 6529 10 88 10 88 0 00 SE Leucocytes 53471 8912 8912 100 00 te Other Leucocytes 0 0 00 0 00 0 00 1938 306 34 3 4 31582 5264 59 06 59 06 3074 512 575 575 16977 28 30 31 75 31 75 D 0 00 0 00 0 00 ere s s ymphocyte vA 264 T lymphocyte Add Population 3 73 26 23 26 Othe Ad enone 470 1 09 a PLAET eon 9 35 63 8 29 RURA 7 5967 13 88 AAE renove Population N 15 23 4 84 is Remove All Populations 9 0 00 0 00 kappa B lymphocyt 989 T 65 38 24 1 85 Lambda B lymphoc 1597 2 66 61 76 2 99 1953 32 11 50 3 65 Gate 0 events Total 0 00 Visibility 0 00 A new window will open where the user can write the name of the function desired and choose the formula Add Function Name CD4 CD8Ratio a CD4 T lymphocytes Partial Type Number Data Population Data Subfunction Population D ta Partial Parameter Data Population cos T lymphocytes vi Other File Selected v Comm
16. nd Dot Plot have been included as new type of graphics that allow to select as many different bands as parameters desired to be represented in the same graphic The number of bands can be as many as the number of parameters in the data file For example in a merged file we could obtain a Parameter Band Histogram with n bands one for each parameter PB HG AAxx KAPPA PE A Log LAMBDA FITC A Log CD24 PE A Log CD5 APC A Log FMC7 FITC A Log CD19 PerCP Cy5 5 A Log CD20 APC A Log CD22 FITC A Log C A Exp SsSc Low FSC A Lin The following Parameter Band Dot Plot shows on red the phenotypic expression of the plasma cells population previously selected max FL4 H CD45 APC Log a Axes Configuration FL3 H CD19 PERCPCY5 5 Log Band 1 FSC H FSC Height I LINEAR a Band 2 SSC H S5C Height Band 3 FL1 H CD38 FITC Y LOGARITHMIC M Band 4 FL2 H CD56 PE Y LOGARITHMIC ael Band 5 FL3 H CD19 PERCPCY5 5 LOGARITHMIC meeen eam eea emam DA FL1 H CD38 FITC Log Band 6 FL4 H CD45 APC iLocaRITHMic M JExp SSC Low v FL2 H CD56 PE Log SSC H SSC Height Exp SSC Lo FSC H FSC Height Lin In some assays it will be very useful to visualize the data with Density Representations since they can help to better distinguish the different cellular populations Version 1 1 of Infinicyt allows visualizing the data as dot plots or density views in different types of gra
17. oels 3 26 MET 12 82 28 95 S K K K K K Gate 0 events Total 0 00 Visibility 0 00 3 1 2 Absolute counts from the population of reference selected by the user To calculate absolute counts there is no need to create the formula The user can choose which population will be taken as reference for the calculation in the Absolute Counts option of the Statistics Window of the Profile Configuration menu Once the population which will be taken as reference for the calculations and the number of event per microlitre of this population is selected the software will automatically show a new column in the Analysis Window with the absolute counts events ul of each population j Pretererces nd mnp Cobo Profile Configuration o Stored Comperasiona 6 Gam JAS Ie Qeni Conpirnshon NepiveV nitty Confess AFS Penne e Propetes Decimal separator Character separator in CSV files Abckie Cores File Statistics Selection Profile Windows Help I Events ma Other Events Monocytes OE EI EI amp oO Gate O events Total 0 00 Visibility 0 00 3 2 Current compensation can be changed manually and saved if the origin FCS file contains this information internally The new cytometric file acquisition programs can save as internal data the compensation of fluorescences used In those cases with the version 1 1 of Infinicyt
18. ofile Configuration menu click Statistics and activate the New Function option A new window will open where the user can write the name of the function desired and choose the formula Profile Configuration lin Preferences Show Statistics i En Mean sD Cy Median Mode New function Controls New function p gt 9 i mm Stored Compensations CWame cosco Raq C c i Gates FCS file J 0 00 H Current Compensation Negative Visibility Configure APS Parameters Value v i Properties 7 Type Number Data Population Data Subfunction Parameter Data Summation Format Decimal separator g Character separator in CSV files Define decimal places 2 Absolute Counts Events pl of the reference population beads Profile Configuration Preferences Show Statistics at Mean v SD Cy Median Mode New function Controls New function 8 Statistics atistic Name CD4 CD8 ratio Stored Compensations i Gates FCS file Current Compensation Negative Visibility Configure APS Parameters Properties Format Decimal separator bs i Character separator in CS files pa Define decimal places 28 Absolute Counts Events pl of the reference population beads Profile Configuration Show Statistics ar Mean sD cy Median Mode New function Cont
19. phs To change to a density view right click on the diagram and select Density Graph Configuration b Density t ow Contour Undo Redo Visibility Window Manipulation ssign Population Population be Configuration b b Percentage of Events Displayed P Invert Axes Graph Info 1 Countour Density considers all the events visualized to perform a density display Fa aaz we Jog Fap Ppa Zon JOa 408 a i 7 4160 a ee 184 SSC HiSSC Haight Exp S5 Low SSC H SS Height Exp 33C Low ae ey ee ee i TEA 402 102 iCal 1 ET VEE VER 14 a AT OR N aes EEA La 2 Population density considers the population already classified to maintain the colours for the density representation The density will be represented degrading the choosen colour ws Sx wn a 2e8 306 406 Gee SOG 10 zee 300 408 GeO S06 PE tee SSC H SSC Height Exp SSC Low SSC H SSC Height Exp SSC Low 181 162 163 ita 162 Tka FLINEEES PEC LeS a l PARES FUSE 08 3 Configuration allows the user to choose the scale of density to show in the graphic Density Configuration Density increment Using the new Automatic Population Separator APS graph in the 1 1 version of Infinicyt the user can choose the parameters to use in order to obtain the best separation of populations The software will take into account both the parameters selected and the rest of events
20. pi a a Population WIS i O 5 Events a Other Events d d L f O d z d T pmphocytes d nE B Lymphocytes d Gate 0 events Total 0 00 Visibility 0 00 Events agog 3117 altata 1654 1552 30075 ar43 17859 1 13044 AEZ 2182 Seles Total Show Events Column Show Total Column 5 one Show Partial Column 94 814 2 rEZ 2 odes ot cies 50 124 SERTE 1ra 10 10 od fb 31 404 0 002 0 012 el fs Taltz 4 ade 14 74 3 644 12 224 Show Visible Column Show Comments Column Show Statistics _14 4 3 The user can configure the mathematical transformation of the parameter SSC to improve the separation of populations In Graph Configuration option the possibility of selecting the values of the mathematical transformation of the SSC parameter to apply 7 Graph Configuration b Axes Configuration in the data file in order to improve the ee x Compensation separation of populations as been added Black Undo Redo oe White Visibility bie se teaenad an naarse The Configurable EXP SSC Low option Window Manipulation W rnin gives the possibility to expand the SSC Percentage of Events Displayed amp show Legend Invert Axes w Show Scale parameter with the values desired by the user When clicking on it the SSC plot will appear with two scrolls in the lower part of the graph to regulate the part of the scale to amplify Graph Info Def
21. re the correct ones the user can save this information The software will ask you for a new name and directory for the new FCS file containing all the parameters merged To go back to the main screen of the program click the Main Menu command 5 2 New features regarding merger process controls See chapter 4 2 4 of the User s Manual to obtain detailed information about Coherence Control to determine if the user is performing the merge process with the optimal files analyzing in different aspects the similarity of expression in the common parameters The version 1 1 of Infinicyt includes the following new features about the merger process controls a The coherence information window about the differences relative to the global coherence control Merge Control has been simplified When the program detects significant 17s differences between the distributions of the events of the different files in the common parameters space it shows a warning message The warning will indicate the percentage of difference value 0 means that there are no differences between the files and value 100 indicates that the distribution of events relative to common parameters is completely different and will also list by significance the parameters and files which make the differences Coherence information Maximum difference between files 26 difference between some files Files and parameters with significant differences FILE 6
22. rols New function Name CD4 CD8 ratio Stored Compensations i Gates FCS file Current Compensation Negative Visibility Configure APS Parameters Properties kd kd Type Number Data Population Data Subfunction Parameter Data Population 2 poe Select Population CD8 T mph v C Other File Selected v m L 4 Population Data Partial Format Decimal separator To Character separator in CSV na Define decimal places 2 Absolute Counts Events pl of the reference population beads Once the formula is finished click OK and the new function will be a new statistic column with values for each population and each parameter ANALYSIS Normal peripheral blood sample Fie Statistics Selection Profile Windows Help Population Partial Mean i 10 88 10 88 659 50 1447 93 Se Leuc ocytes 89 12 89 12 53229 1010 19 Other Leucocytes 0 00 0 00 0 00 0 00 3 06 3 44 131 25 617 14 52 64 59 06 26 95 172 47 5 12 5 75 272 82 831 14 O K K a K K 28 30 31 75 1562 78 1194 11 0 00 0 00 0 00 0 00 ocytes 20 73 2127 68 865 95 Other T lymphocytes 0 97 70 3926 38 1336 96 CD4T lymphocytes 7 39 2122 35 769 34 K 12 37 5967 1989 34 699 17 cytes 431 15 23 16 31 113 44 cytes 0 00 0 00 0 00 0 00 kappa B lymphocytes 1 65 38 24 14 70 138 40 Lambda B lymphocytes 2 66 61 76 17 32 94 73 e NK
23. rt Report Properties Ctrl I Exit Iv K im T lymphocytes Other T lymphocytes CD4 T lymphocytes K K K Si B Lymphocytes Other B Lymphocytes kappa B lymphocytes Lambda B lymphocytes K K K 60000 5707 54293 0 1536 32205 3465 17087 0 12600 687 4262 7651 2563 0 1023 1540 1924 9 51 90 49 0 00 2 56 53 67 5 78 28 48 0 00 21 00 1 14 7 10 12 75 4 27 0 00 1 71 2 57 3 21 9 51 90 49 0 00 2 83 59 32 6 38 31 47 0 00 73 74 5 45 33 83 60 72 15 00 0 00 39 91 60 09 11 26 Gate 0 events Total 0 00 Visibility 4 0 00 The new feature in version 1 1 of Infinicyt is that the analyzed file will be saved in the folder of the hard disk that the user selects with the extension cyt The original data file will remain the same in the corresponding folder because the cyt file does not affect the original one Whenever the user wants to open an already analyzed file then the cyt file must be opened from the option Open Analyzed File of the File menu 4 2 Option to choose the columns to visualize in the Analysis Window By default all columns of the Analysis Window except the statistics column will be visible Nevertheless at any time right clicking on upper columns this information can be hidden or shown ANALYSIS Normal peripheral blood sample File Statistics Selection Profile Windows Help ot
24. sis Window to classify the population in the Population Tree 5 The pre defined gates are part of the current profile configuration therefore clicking the option Gates of the Profile Configuration menu or clicking the option Edit Gates of the Selection menu the software will open a window with options to delete them or change the color of the events and the contour 3 4 Itis allowed to configure negative visibility of the parameters with Logical scale Negative visibility represents the percentage of negative data range in the Logical scale which is shown in the graph It corresponds to the part of negative axes relative to the positive axes Thus whenever the FCS file has been obtained from an instrument with I electronic compensation negative values that are usually close to the axes may be moved to improve the visualization of the positive events Go to Profile Configuration menu and click Negative Visibility to change the percentage shown for each parameter Depending on this percentage the negative part of the scale of the parameter will be more or less represented Profile Configuration Preferences Negative Visibility Percentage Eann Analysis H Graphs i9 Controls C Automatic 1 10000 negativity Statistics Parameter H Stored Compensations H Gates FCS file pocA Current Compensation FITC A CD22 Negative Visibility PE A CD23 roo Configure APS Parameters Per
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