AgileCon Software Manual
Contents
1. Below EQ 4 is Basic parameters Well mapping Quantitative Cut off Ratio Inhibition OC Print options Guiot setings pa cutoff method C Single f Double Ce Calculation Calculate number fa m Reverse Label of all limits Ea1 PC KNO ah bh ch eat EQ a PC b NC c H ita D SE H EQ3 a PC b NC e ah bh ch po2 e04 EQ4 aPC bNC c i e nm EQ4 ne Ratio Inhibition Calculation Method Select a BO as a standard value to calculate the rest of the well plate Bn Basic parameters Wel mapping Quantitative Cutoff Ratio inhibition le Fatio Inhibition C Ratio B50 Definition of BO 1 Ratio Inhibition operating procedure a N b C N d O definition Ratio Bn B0 Inhibition 100 Bn B0 Must have sample on BO position or the AgileCon will show error Please check if BO is a replicate of the sample if there are many replicate of the sample the BO will be the average of the replicates of the sample If BO value is 0 then the AgileCon will show error If ratio is over 200 then the AgileCon will show HI lower then 200 then the AgileCon will show LO Ratio is represent as blue Inhibition represent as red 38 Q C Calculation Method The criteria of the QC calculation can used to determines on the reliability if this experiment 1 SS sl Can use a maximum of 4 equation for calculation all of the calculation2. File Experiment Setup Help op S pr A I ES OFF LINE Basic parameters Well mapping Quantitative Cut off Ratio Inhibition oC Print options Parameters Measurement type Two Points l m Filter wavelength nm Incubation EE Previous Current Use incubator 5 Sr Maint 450 450 Temperature i pete jw Ratio lnhibition Main2 0 o is je vac Iw Reference filter skar S Ref1 0 KH M Use shaker Two pointinterval 70 a ser Ref 2 fo 450 Starting method Ze Immediate C Delay Plate motion amp Continuous Measurement unit C Stepping Unit mgidL 11 e Kinetics read During a kinetics read users can define the kinetic method by select the Kinetic method tap User can define average rate Maximum rate Maximum Abs Total delta Abs Time to max rate and Time to max Abs User can also define the number of times the plate been read and intervals File Experiment Setup Help Para ean Basic parameters Well mapping Quantitative Cut off Ratio Inhibition Q C Print options Parameters Measurement type Kinetics Filter wavelength nm Incubation 7 Quantitati Previous Current M Use incubator S Sea Maint 450 faso xi ope Ratio nhibition 15 e MV Qc Shaker Use shaker Starting method d Kinetic method Average rate fe Immediate 3 Average rate easure numbers freien
3. Select COM port pa 8 78 AgileCon Menu Software Structure Main Window Overview i File Experiment Setup Help A B a aE Basic parameters vell mapping Quantitative Cut off Ratio Inhibition Q C Print options Parameters Measurement type End Point D m Filter wavelength nm Incubation en Previous Current E Use incubator Se Maint 405 v Temperature p Je Ratio Inhibition 15 ebe VM QC Iv Reference filter Shaker Ref 1 490 bd W Use shaker Speed Ge Low 8Hz r Starting method C Mediumt11Hz Immediate C High 14H2 A Menu Time B Tool bar Ge Dela d 0 se ser C Message D Temperature monitor Plate motion E Status monitor Measurement unit C Continuous F Working area Stepping 0 ms ME G Desktop bar 030J04d Section A Menu Section B Tool bar Section C Message Section D Temperature monitor Section E Status monitor Section F Working area Section G Desktop bar ne Section A Menu The File Menu contains function for processing data from the AgileCon G Experiment Setup New Load B4 Save ative Cut off Ratio Inhibition Q C Print options Save As Generate Preview report Export Print report incubation Exit armer h Quantitative SS se incubator M Cutoft Temperature h Ratio nhibition 15 sje v ac R
4. properties of any device Drivers Driver Signing lets you make sure that installed drivers are compatible with Windows Windows Update lets you set up how Windows connects to Windows Update for drivers Hardware Profiles Hardware profiles provide a way for you to set up and store different hardware configurations Hardware Profiles Figure A 1 47 ne Device Manager File Action View Help e EI S E A Sa SL user Computer De Disk drives m S Display adapters 2 DYDICD ROM drives He Floppy disk controllers L Floppy disk drives 41483 IDE ATAJATAPI controllers H Keyboards H t Mice and other pointing devices E 3 Monitors S Network adapters JN Ports COM amp LPT Ei Communications Port COM vee Ei Communications Port COM ECP Printer Port LPT1 a USB Serial Port COM3 1 g s Processors GR Sound video and game controllers b System devices cs Universal Serial Bus controllers Figure A 2 USB Serial Port COM3 Properties General Port Settings Driver Details Bits per second arsil Data bits Parity Stop bits Flow control Advanced Restore Defaults Figure A 3 48 F ig ure A 4 49
5. exp file AgileCon SW will check Save Save experiment file Save As Save experiment file using different file name Close Close experiment file Generate Preview report Generate report after experiment is done must generate report first before saving to Excel 6 Export Print report To export to excel or to Print report using the printer connected to the PC 7 Exit Exit the AgileCon SH dl se PX ne Experiment menu functions There are 2 functions under the experiment menu Protocol and Data Protocol is to setup the parameters of a experiment and data is to show the results of an experiment 1 Protocol To setup the Parameters Well mapping Quantitative Cutoff Ratio Inhibition Q C and print options of a experiment File Experiment Setup SE or ISj m elt ao Basic parameters well mapping Quantitative Cut off Ratio inhibition Q C Print options Parameters Measurement type End Point MM Filter wavelength nm Incubation 09201014 Quantitative main Use incubator Cutoft Temperature A IT Ratio nhibition 15 Gre Reference filter Shaker Use shaker Starting method Ce Immediate gje Delay Plate motion N Measurement unit Ze Continuous Unit None r 2 Data To show the result of an experiment Stepping T File Experiment Setup Help gt S F 8 i el TA l Tees Bi fees
6. 0 231 0 144 0 105 0 139 0 120 0 204 0 242 1 102 1 051 1 123 1 057 1 119 2 090 1 119 1 071 1 085 1 062 1 183 1 262 1 229 1 252 1 332 Conc 120 000 145 000 1000 000 1200 000 Calib curve Fit type cubic polynomial Meas scale linear Conc scale linear 0 0 1575 Formula Conc ABS a0 al C a2 C C a3 C C C al 2 954E 1 al 3 712E 3 a2 3 476E 6 a3 1 137E 9 Layout Shows the well mapping layout of the plate Different types of well uses a different color to represent source data Shows the source data for the quantitative measurement e In end point measurement if there is no reference filter then the main filter M1 data is the source data If there is reference filter then M1 R1 is the source data e In Two points measurement if there is no reference filter the source data will be M1 e In Two points measurement if there is reference filter then the source data will be D1 M1 R1 e During Kinetic measurement user cannot use reference filter the M1 data will be the source data Calibrators Will use CO1 C15 to represent every STD s name and OD value and will show the average measurement and the standard Conc value Calib Curve When using standard curve Curve on plate or stored curve will use curve fitting to create a standard curve Residuals table Will use C01 C15 to show standard OD value C set Average Abs and Concentration and Re
7. 5 1 POSOL STDOL sToo2 sToos STDOG STDOS E SE 1 2 02 72 0 3 2 C4 2 0 5 2 POSOL STDOL STDOOZ STDOS STDOG STDOS a 21 1 01 31 02 31 03 3 c4 3 C 5 3 BLEOL STOOL sTooz STDOS stood STDOS acre Bem MENNENE SR NEGO1 STDOL STDOZ STOO STO OG STDOS Meqative Standard NEGOL STOOL STDO STDOS 3TOO4 STOOS Type Standard Fill direction Replicate direction Mame ISTO Ce Row Ce Row Fill number fi C Column Canc 32 000 al C Column Replicate number fi 2 Well mapping method a Select the type well users would like to define Blank Positive Negative Sample Standard Users can also enter the concentration value for the standard b Determine where on the well the well type needs to be and left click on the position on the well c Right click on the mouse to select the fill option 3 Fill and replicate direction rule Row is to fill or replicate the well vertically column is to fill or replicate the well horizontally 30 PE ne 4 Fill and replicate well rules a Fill number Is to replicate the number of different samples b Replicate number Is to replicate the sample how many times on the well plate Example of filling and replicate the well plate 12 13 2 18 5 Blank POS and NEG can only have 1 set within AgileCon 6 STD can configure 1 15 set a maximum of 96 7 Sample max of 96 8 Types of well a BLK User can
8. STOOL STOO STDOS STDO4 STOOS Z 1 1 E Tea EE GL 3 3 C 4 3 iS 5 3 BLEOL STOOL STOO STDOS STOO4 STOOS M 1 1 LC 1 4 2 4 C 3 4 L 4 4 I 5 4 MEGOL STOOL STDOOZ STOOS STOO4 STOOS fl 1 2 C 1 5 GE 2 5 EG 4 5 is 5 5 MEGOL STOOL STOOs STDOS STOO4 STOOS Type Sample d Mame Gau Fill direction Ce Row C Column Replicate direction ng Rae Fill number fi GC Column Replicate number fi 14 Quantitative The AgileCon allows user to define quantitative analysis to determine the absolute or relative abundance There are 4 different types of curve fitting on the AgileCon Users can use Curve onplate Stored curve Standard line and Concentration factor To define Quantitative analysis 1 Make sure the check mark is clicked on the Quantitative in Basis Parameter 2 Click the Quantitative tap in Basic Parameter 3 Define the desired parameter TV File Experiment Setup Help Odal NE Buantitative settings Curve on plate Data curve fit x scale v scale e Li Sat Cubic polynomial Linear e Linear C Logit Login C Stored curve C Standard line f Concentration factor 15 Cutoffs Cutoffs are used to classify results Users can define 3 different Cutoff methods Single Double Calculation 1 Make sure the check mark is clicked on the Cutoff in Basis Parameter 2 Click the Cutoff tap in Basic Parameter 3 Define the desired parameter TV File Experiment Set
9. Two Points Filter wavelength arr Previous Current Maint 450 Bi M Mainz 450 405 hd Reference filter Refi ken faso gt Ref2 aso Jeng Starting method Incubation Use incubator Temperature Shaker f Use shaker Speed Ce LowiSHz C Medium iHz Highil 4Hz Time i SEL C Immediate Ge Delay m SEL Flate motion C Continuous Measurement unit re Stepping 0 ze bai maid 97 Mo Quantitative W Cutoff h Fatiolnhibition M aC h Quantitative W Cutoff M Fatiolnhibition 15 C DC Two point interval z pe SEC C N reference filter Farameters Weasurement type kinetics Filter wavelength nm Previous Current Maint faso 405 Starting method C Immediate Ce Delay p 4 SEL Plate motion Continuous Ge Stepping Incubation Use incubator Temperature fs He Shaker W Use shaker Speed Ce LowiSHz C Medium 1Hz C Highil 4Hz Time Nr SEL Measurement unit be mod Kinetics Kinetics measuring method can only select main filter and no h Quantitative f Cutoff M Ratiolnhibition W aC Average rate Maximum rate Maximum Abs Total delta Abs Time to max rate Time to max Abs kinetics method Average rate Measure numbers E E Sac Measure interval Filter wavelength Users will need to select the filter wave
10. lt 1 200 S OC3 0500 lt 0000 PC 1000 NC 0 000 lt 1 000 Q Pass condition 10 if oc TRUE then PASS 11 OC DCL AND OCH 12 Result Le OC1 FAIL 14 OC3 PASS 15 oc FAIL 16 Kinetic results When using the kinetic measuring method AccuMate will display the kinetic curve of every reading User can check the reaction rate easily on this screen mo xj Well B6 scale range Sampling numbers 5 1 766 max fo in well BG scale range Sampling numbers 5 1 766 max Exit 45 ne View Report To view report users will have to go into file and click the Generate Preview report tab for the AgileCon to generate report After reports being generated users can use the Export print option to save to Excel or print The report will be saved in Program FilesVACTGenelAgileCon Data under excel file MO os fl fr E C lt Experiment tHe gt Name User Mote Date 06 10 08 lt Protocol parameters gt Experiment file path Measurement type Measurement mode Main 1 filter nm Starting method Need shake Weed incubate Need quantitative Quant method Quant standards number Quant replicates number Curve fit method Cutoff method Upper threshold label Lower threshold label Need ratio inhibition BO well Need quality control OC1 O C3 Pass condition Calibration Ti
11. rate C Delay Maximum Abs Measure interval Total delta Abs Plate motion Time to max rate j Time to max Abs arne Measurement unit C Stepping Unit mg dL To select a measurement type Click the tap under measurement type and select the desired method of End point Two points Kinetics Define Primary and Reference wavelengths lf a Primary wavelength is defined alone the AMPR 750 reads the plate only once ata single wavelength If a Reference wavelength is defined the plate will be read twice and automatically to calculate the delta Abs between the two readings To select the Primary and Reference wavelengths 1 Select the Measurements type of End Point and Two Points 2 Enter the Primary wavelength in Main1 or Main2 and the reference Ref1 or Ref2 Starting method to read plates Users can define the starting method of plates reading Immediately the AMPR 750 starts reading the plate right after executing tap is pressed Users can also define how 12 ne long the plate reading delays To define the starting method 1 Click on the immediate tap or define how many seconds needed to be delayed Plate motion Users can define the plate motion while reading the plate to be Stepping in milliseconds or Continuous Using the built in Incubator The incubator will set the temperature of the plate at ambient temperature Users can enable the Incubator by 1 Click the use incubator tab 2 Enter the desire
12. AGILECON SOFTWARE USER MANUAL Version 1 4 ACT Gene Inc provides this publication as is without warranty of any kind either express or implied including but not limited to the implied warranties of merchantability or fitness for a particular purpose Information in this publication is subject to change without notice and does not represent a commitment on the part of the vender Any errors or omissions which may have occurred in this publication despite the utmost care taken in its production will be corrected as soon as possible but not necessarily immediately upon detection ne Contents System Requirements rrronnrnnnnnrnrnnnrvrnnnrnrnnnrnnnnnennannennansennnnnnnnansennnnsennnnsnnnnssnnnnnee 1 MN 2 PEN 3 AgileCon Menu Software Structure seese RE E 4 Main Window Overview rrrrarenarennnrrnanenanennnrnnanenanennnnnnanenanennnsnnanennnennnnnnnnennn 4 MC CHOMP On UE 5 SMP NE E 8 SEC CG MESTE vr 8 Section D Temperature monitor 8 SUE SN 8 SF NNN 9 SE GDS Barr 10 AEG ON Function wjecratcenicnaincmscnainendieandeendicendeandenaiaeetoanieseisentessdeceinenteaniocteerteandeosdaes 11 Etienne 11 Define Primary and Reference wavelengths nnunnennnnnnnnnennenennennrnrenennnne 12 SlaMING method to fe Be EE 12 Plate motion ee 13 USING EP re 13 VENTE ei En Ee 13 Me EG 13 Define Calculation rorrrrrnnrrrrnnnrrnnnnrvrnnnenrnnnrnrnnn
13. Quantitative Cut off Ratio Inhibition oC Print options Parameters Measurement type Kinetics Filter wavelength nm Incubation ae Ww Quantitative Previous Current MG Use incubator h Cutoff Wain 450 o Temperature i w Fatiollnhibition hs Se w HL Shaker h Use shaker Speed e LowtsHz due C Medium 11H2 Kinetic method Average rate Ge Immediate C Hight Aa REFERER C Delay Time Measure numbers Meine 1 ser Measure interval Ma4mum Abs SS Total delta Abs Plate motion Time to max rate Measurement unit Time to max Abs f Continuous f Stepping nit ih a Kinetic method Uses can select Average rate Maximum rate Maximum OD Total delta OD Time to max slope Time to max OD 29 b Measure number User can enter the measuring numbers of the plate from 3 30 times c Measure interval User can select the measure interval When plate motion is in Continuous from 4 500s in Stepping 5 500s Well Mapping 1 Save and load map layout Users can load their map layout by pressing the folder under map layout or the disk icon to save under map layout Map layout CIS Select type firstthen assign location to fill 10 11 12 Ts Petre re est Ed ap tales Tien SAMOS samog SAMO7 SAMOS SAMOS SAMLO Samii SAM12 se NNN SE 7 samos samoa samio S4mii SAM12 im STRD T7623 S23 2 tees i ties Tr SEN ar 7 samos samoa samio S4mii SAM12 DERE ii 2 1 3 1 caa
14. ameters When starting experiment users must first define the parameters such as wavelength reading method incubator and shaking In addition users can also define the calculation such as Quantitative Cutoff Ration Inhibition and QC 1 Measurement Type Users can define 3 types of measuring types End point two point and Kinetic Basic parameters Well mapping Quantitative Cut off Ratio fInhibition oC Print options Parameters Measurement type Two Points End Point Incubation Use incubator Temperature h Ratio lnhibitian 15 C DC Shaker f Use shaker Two point interval E SEL Speed Ce LovwiSHz C Medium iHz Hight 4H Time fi SEL Measurement unit Unit ei h Quantitative W Cutoff Starting method Ce Immediate C Delay Plate motion Ce Continuous Stepping 6 Ad 8 78 a End Point Parameters Measurement type End Point Filter wavelength nm Previous Current Wain 450 405 W Reference filter Ret 1 0 490 Incubation Use incubator Temperature fs He shaker h Use shaker Speed Ge Latz C Medium Hz C Hight AH Time a SEL Starting method C Immediate fr Delay Dr Sec Plate motion Measurement unit C Continuous re Stepping fo 4 EE Unit None b Two Points Parameters Measurement type
15. are version of the AgileCon In the initial setup the AgileCon will scan all available ports if there is no response from the AgileCon the AgileCon will show off line 19 vg 8 9 Select the desired COM port for the communication between the PC and AMPR 750 Currently the AgileCon supports COM 1 COM16 x Press OK on the AgileCon to start communication between AMPR 750 and AgileCon Press Init on the tool bar of AgileCon The AgileCon will use the desired COM port and send a signal to the AgileCon If the AMPR 750 responds to the LCD the AgileCon will show ON LINE Setting up USB connection Between AMPR 750 and AgileCon 1 2 On AMPR 750 standalone mode go to SETUP3COMPUTER Change the RS232 to USB Power on the AMPR 750 again while pressing option button on the instrument to switch to the PC mode Appendix A for USB setting 20 Main Menu Configuration Graphic User Interface The graphic user interface will have the function menu tool bar message status Temp connection status and working area Protocol and data Experiment Setup New Load Save Save As Generate Preview report Export Print report Exit File Experiment etup vw Protocol Data File Experiment Setup COM port setting Filter tone File menu functions Under the main menu of the AgileCon there are four options Files Experiment Setup and Version 1 Load Load experiment file When loading
16. bet ee er She i a eae L ee N 2 E E Ee Ei KE ER Experiment file path Sample exp easuremen e End point Continuous 030 01d 450 Immediate N N Yes Curve on plate 5 umber 5 Cubic polynomial Double threshold 9 Upper threshold label for gt 2 000 Lower threshold label for lt 1 200 eq Need ratio inhibition Inhibition 10 BO well C10 19 Heed quality control Yes QC1 1 000 PC 1 000 NC 1 500 lt 1 200 21 Qc3 0 500 lt 0 000 PC 1 000 NC 0 000 lt 1 000 Pass condition OC OC1 AND OC3 22 one Setup menu functions The setup menu is used for COM port configuration and filter setting The AgileCon can setup up to 8 filters 1 COM port setting User can change the desire COM port to communicate between AMPR 750 and AgileCon AMPR 750 will automatically detect all available COM port on the PC COM port setting 2 Filter tune The AgileCon has one 8 slot filter wheel for the users to install filters After installing new filters on the AMPR 750 it is important to set the correct filter wavelength on the AgileCon Check the check box on the left to enter desired wavelength for the filter and press the Tune key after the desired wavelength is entered Filter amp lamp tune xX Fiter wavelength settings Fiter MoO Fiter Mod Jags Filter No 5 Filter Mos 5 Fiter Mo 4 1490 Filter Mo 5 oc PE ne Tool Bar Menu C
17. c parameters Well mapping Quantitative Cut off Ratio Inhibition c Print options Frint options Title setting Mame User Mote Sections for printing h Title P OC validation i Parameters Ca Calibration if Data list h Layout WwW Raw abs matrix h Cutoff matrix vw Blanked abs matrix mw Ratioilnhibition matrix 18 ne AgileCon connection setup To control the AMPR 750 through the AgileCon users must set and test the COM port connection on the PG User can connect the AgileCon with AMPR 750 through USB or RS232 interface Start AgileCon under PC mode 1 2 5 Attach the appropriate cable to an available RS232 serial port or USB port Plug the other end of the cable into the RS232 serial port or USB port on the reader Plug all power cords into electrical outlets Power on AMPR 750 if AMPR 750 is in standalone mode power off while pressing Option button to switch to PC mode Configure the communication COM port between AMPR 750 and the PC Setup Connection between AgileCon and the PC 1 2 Ensure AgileCon is connected to the computer Ensure on the AgileCon LCD shows is in PC mode Ensure the COM port setting is Baud rate 57600 Data bits 8 bits Parity check no Stop bits 1 Flow control no Go to start gt program files gt ACTGene gt AgileCon to execute the AgileCon software After executing AMPR 750 the PC will show the startup screen with the softw
18. ce filter Ref 1 490 z Starting method C Immediate Delay p 3 sec Plate motion C Continuous Stepping fo ms Incubation Use incubator Temperature fs Ze Shaker W Use shaker Speed Ge Low 8Hz C Medium 11Hz C High 14Hz Time 1 SEL Measurement unit pre MW Quantitative Jv Cutoff W Ratiollnhibition V QC 030J04d A Menu B Tool bar C Message D Temperature monitor E Status monitor F Working area G Desktop bar PE ne Section G Desktop Bar The desktop bar is to select between the Protocol and the Data this function can also be accessed under experiment Bd POR ee shop Da E Ls Bor gt B lt 10 PE ne AgileCon Function Basic Parameters e Measurement types The AgileCon provides 3 types of measurement types end point two point and Kinetics File Experiment Setup Tel else F L E Seck Basic parameters well mapping Quantitative Cut off Ratio Inhibition O C Print options Parameters Measurement type oO Incubation Ka 7 Quantitati Use incubator e 8 SE M Cutoff Temperature one E Ratio nhibition 15 C Eac Reference filter ES Shaker Use shaker Starting method Ce Immediate e bi C Delay e Two points read During the two points read theAMPR 750 reads at 2 wavelength with 2 reference reads as optional
19. d temperature Using the built in Shaker The built in Shaker in AMPR 750 can let user define 3 settings low 8Hz Medium 11Hz and High 14Hz Users can also define how long the shaker needs to be active by seconds To enable the Shaker 1 Click the Use shaker tab 2 Define the speed Low Medium High 3 Define the time in seconds Measurement unit Users can define the Measurement unit to suit their experiments needs To define the Measurement unit 1 Click on the unit tab and select the desired Measurement unit Define Calculation User can define Quantitative Cutoff Ratio Inhibition and QC calculation method To define calculation method 1 Click on the tab in respect to Quantitative Cutoff Ratio Inhibition and QC calculation 2 The AgileCon lets users define each calculation method criteria i ne Well Mapping Users can define 5 types of different wells Blank Standard Sample Positive and Negative To define Well Mapping 1 Click on the Well mapping tap on the main working acres 2 Users can define Blank Standard Sample Positive and Negative for each well Si File Experiment Setup Help SEIN Eas Basic parameters Well mapping Quantitative Cutoff Ratio i Inhibition oC Print options hap layout f Select type first then assign location to fill F1 1i C 1 1 C 2 1 ES CG 4 1 5 1 POSOL STOOL STOO STDOS STOO4 STOOS Pi C 1 2 T 2 2 C 3 2 LC 4 2 L 5 2 POSOL
20. define BLK for the blank well and are defined as a light green on the well plate b POS Uses can define positive control on the well plate and are defined as a light red on the well plate c NEG Users can define negative control on the well plate and are defined as a light blue on the well plate PE ne d Sample Users can define the sample on the well plate and are defined as an light orange on the well plate e Standard Users can define the standard on the well plate and are defined as a light purple on the well plate Users can fill in the value of 0 001 999999 999 TV File Experiment Setup Help Wap layout f Select first then CC location to fill ee En EM ae SE LEN EEG BLEOL BLEOL BLEO1 BLEO1 BLEDI pe bales e das oP oe PIS POSO1 POSOL POSO1 POSO1 POSOL N 1 1 Ni 2 Ni1i 3 MIA ML 5 NEGO NEGO NEGO NEGO MEGOL ne EE C 3 1 3 el CH En cl E l 3 Se STOO STOOL STOOL STOOL STOOL Fill direction C Row Ze Column Replicate direction ag Fav Fill number fi C Column Replicate number 5 Mame 3 Quantitative measuring method AMPR 750 supports 4 types of Quantitative method Curve on plate Stored curve Standard line and Concentration factor 1 Curve on plate Uses the standard on the well plate for the calibration curve There are 7 types of curve fitting on the AgileCon a Linear regression b Qua
21. dratic polynomial c Cubic polynomial d gt Point to point e Cubic spline IT 2 parameters logit log g 4parameters logistic Basic parameters Well mapping Quantitative Cut off Ratio Inhibition oC Print options Huantitative settings scale Ge Linear C Logit Scale fr Linear C Logit f Curve on plate Data curve fit Cubic polynomial Linear regression C Stored curve iuadratic polynomial Cubic polynomial Point to point Cubic spline Standard line 2 parameters logitlag 4 parameters logistic C Concentration factor 33 Ad 8 78 2 Stored curve Users can load their existing stored curve for quantatitive the stored curve are under AgileCon StdCurve file name is cuv Basic parameters Well mapping Quantitative Cut og Ratios Inhibition oc Print options Uantitative settings C Curve on plate Stored curve C Standard line C Concentration factor 3 Standard line User can use the Abs A Conc B equation and enter the value of A and B to calculate a standard line The Value of Acan be 999999 999 999999 999 The value of B can be 999999 999 999999 999 Basic parameters Well mapping Quantitative Cut off Ratio Inhibition OC Print options Huantitative settings C Curve on plate Stored curve Abs A Conc B A fi B fi C Concentration factor oO PE ne 4 Concentration factor User can enter a factor for calcu
22. eference filter a Shaker Use shaker Starting method e Immediate f Delay Plate motion 8 Measurement unit f Continuous f Stepping Unit mgidL New open new file window Default file name is new exp Load Load an existing results record Save Save the experiment parameters save As Save the results under a new ID Generate Preview report Generate experiment report in data Export Print report Export report to excel file and select to print report Exit Close the AgileCon software PE ne e Experiment Menu Experiment Menu contains function to set the experiment and view the data pr 4 OFF LINE Basic parameters well mapping Quantitative Cut off Ratio Inhibition O C Print options Parameters Measurement hype End Point Fitter wavelength nm Incubation SEN h Quantitative Previous Current Use incubator F Cuon Maint 450 Temperatu ae ain 0 Geer F Ratiofnhibition 115 k WRC Reference filter Shaker Use shaker Starting method Ze Immediate C Delay Plate motion Measurement unit f Continuous Stepping Unit m tt Protocol Set the environment Data View data for the experiment The Protocol and Data function can also function on the right tab of the main menu File Experiment Setup Help FX IQ F adi OFF LINE 5 Basic parameters Well mapping Quantitative Cut off Ratio inhibition O C Print opti
23. ennnnsennnnnennansennansnnnnnsnnnnssennnn 13 My CII a e ere D 14 STE EO EEE NE NN E 15 te EE 16 ae ULL OA 0101 0 0 0 PDS 16 Oo EE 17 0 019 SE E 18 AE ON connection SEP Lean 19 Start AgileCon under PC mode rrrrnnnnnnannnnnnnnnnnnnennnnnennnnnennannennnnsennnnsnnnnneene 19 Setup Connection between AgileCon and ebe 19 Setting up USB connection Between AMPR 750 and AgileCon 06 20 MEN Menu CMD vvs 21 Graphic User Interface oannnannnnannnennnnnnnnennnrnnnnrnrnnnnrnrrnnenrrnnrrnnnrrsnrrrerernnni 21 EET OIG ee 21 EMNENE TN ek 22 DSU Me MU Ua os NN E 23 Tool Bar Menu Configuration ccccccccsecccscecesceteuceceneeseuceseuceseueeseueeseueesaueesenees 24 ne Defining Parameters for Experiment Defining Parameters Well Mappmg Quantitative measuring method Cutoff measuring method Ratio Inhibition Calculation Method Q C Calculation Method Printing opttons ssisrossicsiissrdariiis Interpreting the results Appendix A Setting of USB Serial Port System Requirements 128MB of RAM above for Windows XP Windows Vista Windows 7 Microsoft Office 2003 Microsoft Office 2007 Microsoft Office 2010 20MB of available hard drive for the program files CD ROM drive 16bit color display 800 x 600 Keyboard mouse and RS232 serial port or USB PE ne Software Installation To ins
24. late the concentration The value of F can be 999999 999 999999 999 235 PE ne Cutoff measuring method AMPR 750 supports 3 types of Cutoff measuring methods 1 Single cutoff method User can enter a threshold of 0 0000 4 0000 and define the result is positive or negative Basic parameters Wel mapping Quantitative Lut off Ratio J Inhibition OC zuto setings Cutoff method f Single lf result threshold then Threshald fo Positiver Cutofflabel f eg mme C Double C Calculation 2 Double cutoff method Users can define the high and low threshold The high and low value can be from 0 0000 4 0000 The AgileCon can determine the positive negative results and between Basic parameters Wel mapping Quantitative Cut off Ratio Inhibition O Cutot setings lt Aaa Cutoff method C Single High threshald 2 If result threshold then C Positivt Low threshold 1 2 e Ge Negative Cutoff label Calculation 36 3 Calculation cutoff method Users can create a maximum of 4 formulas as the threshold calculation and make the result as 5 groups Every formula can be calculated from the positive or negative control results with the equation a rb C The value for a b and c can be 1000 000 1000 000 Example With four threshold the threshold higher then EQ1 as default Between EQ1 and EQ2 Between EQ2 and EQ3 is Between EQ3 and EQ4 is
25. length for the desired experiment In addition users can also select a reference wavelength AMPR 750 will automatically calculate the Delta OD as for difference between the main and reference filters Starting method Define when to start the defined experiment parameters a Immediate Start right after pressing the execute tab b Delay Users can define from 0 999s delay to start Plate motion To define how the plate motion when measuring a Continuous When measuring the plate is in continuous motion and not stopping D Stepping User can define the stop time when measuring from 0 999 ms In kinetic mode there is no stepping time Incubation Users can define the incubator temperature by clicking the incubation tab AMPR 750 incubator can save the temperature from 3 50 C oe 6 Shaker The shaker onAMPR 750 can select 3 types of speed and the shaking time can range from 0 999s 7 Measurement unit Users can select 15 types of measurement unit None G dL U L G L ug dL ABS mg dL l OD mABS U mL ug mL mEq L mmol L umol L ng mL When select to None user can enter the desired measurement unit 8 Two point interval Users can select the 2 point interval from 5 999s 9 Kinetic method numbers and interval When user select kinetic measurement user can select the method numbers and interval Di File Experiment Setup Help S ome I Basic parameters Well mapping
26. me 17 06 44 Sample exp End point Continuous 450 Immediate No No Tes Curve on plate 3 3 Cubic polynomial Double threshold for gt 2 000 for lt 1 200 Inhibition Cio Tes 1 000 PC 1 000 HC 1 500 lt 1 200 0 500 lt 0 000 PC 1 000 HC 0 000 1 000 QC OC1 AND QC3 46 Appendix A Setting of USB Serial Port If your Windows XP 98SE PC has USB ports only please install the attached USB to RS232 driver on PC Connecting the USB cable between PC and AMPR 750 then power it up Please set up your Windows XP computer as follow Under Systems Tasks of My Computer screen press View system information In System Properties select icon Hardware and press button Device Manager figure A 1 In Device Manager double click USB Serial Port figure A 2 to enter port setting figure A 3 Please remember the com port number for further use in appendix B Fill in 57600 8 None 1 None in those blanks Press button Advanced to enter advanced COM seiting figure A 4 USB transfer size is to be selected as max number 4096 Latency timer is to be smallest number 1 Upon finishing above procedure please go to appendix B for further operation System Properties HME System Restore Automatic Updates Remote General Computer Name Hardware Advanced Device Manager The Device Manager lists all the hardware devices installed on your computer Use the Device Manager to change the
27. onfiguration There are Init Execute Stop COM port setting Plate In Out Shaker Incubator and post processing tabs on the tool bar menu 1 Init tab After connecting AMPR 750 and AgileCon users must initialize first before any changes can be made l S FI e Exec tab To execute the desired parameters for the experiment OSO GI Stop tab To stop the current action for the AMPR 750 OLAS Baal OF SRS COM port setting To setup the COM port connection between the PC and the AgileCon Plate in out To open close the plate the plate status will show on the status bar Shaker The shaker tap is used to configure the shaker The shaker has 3 speeds low 8Hz Medium 11Hz High 14Hz O gt S 7 ms Incubator To initialize the incubator users can configure the incubator from ambient 3 50 C OP S Post Processing Use the parameters on the main manual and refresh the data Q 8 I D 9 Message Shows the status and messages from AMPR 750 INIT 10 Temperature monitor To monitor the incubator temperature within AMPR 750 Plate state 31 3 i C Connection AI Status monitor To monitor the status of AMPR 750 After initialization there will be two lights showing on the status monitor Red Green represent plate is Out In Blue Grey represent connection status between AMPR 750 and AgileCon 5 Defining Parameters for Experiment Defining Par
28. ons Parameters Measurement type End Point D Filter wavelength nm Incubation Previous Current TC Use incubator s dion 8 1 eso T f de F RanoAnhiben s W OC M Referente filter Shaker Use shaker Starting method G immediate g C Delay Plate moton Sta Measurement urit C Stepping Vent mgl D e Setup Menu The setup menu contains the AgileCon system configuration File Experiment COM port setting E Filter tune Basic parameters Wel l mapping Quantitative Cut off Ratio Inhibition OC Frint options Farameters Measurement type End Point Filter wavelength nm Incubation fas M Quantitative Use incubat Main Use incubator Cutott Temperature EE Ratio lnhibition Bt pas Reference filter EZ Shaker J Use shaker Starting method f Immediate f Delay Plate motion Measurement unit f Continuous C Stepping Unit None COM port setting To set the communication COM port between the AgileCon and PC Filter tune The AgileCon can setup to 8 different filter wavelength Config the filter wavelength ne Section B Tool bar gt S F Be el Post processing Incubator Shaker Plate in out COM port setting Stop Execute Initialize i Initialize Any commands issued from the AgileCon to the AMPR 750 must initialize first Execute To start the defined protocol Stop To stop executing the defined protocol COM port setting To se
29. result are considered to determine pass or fail of the QC calculation method The value of a can be 1000 000 1000 000 The value of b can be 1000 000 1000 000 The value of c can be 1000 000 1000 000 The value of H can be 9999999 999 9999999 999 The value of L can be 9999999 999 9999999 999 To determine pass or fail the queation uses a OR AND XOR to determine Basic parameters Well mapping Quantitative Cut off Ratio t Inhibition Dt Print options BEIS General equation L aPC DNG c H L a h H W aCi Mia si P4 fi hC 15 JIE WM ac 0 sl PC fi SEA 0 D d NIT Pass condition CH QC true then PASS I fil OC Jor IR AND EISE 39 Printing options Users can set Name User and Note to differentiate different experiment reports Users can also check the boxes next to the sections for printing Colum to determine which data users will need to show on the report Pe Dil eS a 9 Basic parameters Well mapping Quantitative Cut off Ratio Inhibition Q C Frint options Print options Title setting Mame Leer Mote x Sections for printing jv Title P Delta ahs matrik M DC validation W Parameters f Calibration jf Data list f Layout Kinetics f Cutoff matrix W Blanked abs matrix h Ratiollnhibition matrix Title The title of the experiment Parameters Shows the protocol parameters of the experiment Layout The well plate layo
30. sidual Ccal Cset oF lo PX ne Curve Viewer User can double click on the curve to enable the curve viewer User can also store the curve in the save curve tab on the bottom The default stored curve are in AgileCon StdCurve Curve Yiewer F log Abs 0 30 2 00 2 00 3 20 log Conc ABS al a1 C a2 C C a3 0 C0 C al 2 954E 1 al 3 712E 3 Save Curve a2 3 476E 6 a3 1 137E 9 Data sheet According to the well plate mapping the AgileCon will display all the result The data sheet will show Name Well ID Replicate numbers Abs SD CV Conc Measuring unit Cuttoff and Inhibition The average of every value will display a avg next to the well ID Results Data Cute Rationnhibiton oc L Knete Y Repon Name Well Replicate avs 8D cv Conc Unt cuion inners P9501 D4 1 0 022 POSsO1 E4 2 0 039 EE NEGO1 G4 1 0 032 NEGO H4 2 0 115 SAMO1 Ad 1 0 036 SAMO1 Di 2 0 112 SAMO C1 3 0 100 SAMO A2 1 0 029 SAMO B2 2 0 071 SAMO C2 3 0 014 SAMOS AS 1 0 006 ae Sie SAMOS B3 2 0 003 SAMOS C3 3 0 010 e lt SAMO4 Ad 1 0 120 Se SS a SAMO4 B4 2 0 048 D SAMO4 CA 3 0 041 SAMOS AS 1 0 280 SAMOS BS 2 0 167 SAMOS C5 3 0 158 ve SAMOG AG 1 0 292 AE SAMOG BE 2 0 222 ER SAMOG CE 3 0 159 sl Cutoff results When clicking
31. t the COM port to communicate between the AMPR 750 and the PC Plate in out To open or close the plate compartment Shake To start the shaking function on the AgileCon Incubator To manually start the incubator when incubator is initialized the temp will show on section D temp monitor Post processing Use the current protocol to re process data results Section C Message During operation the message will show on the right side of the tool bar INIT Message text Section D Temperature monitor When incubator is active the temperature will show on the right side of the tool bar Section E Status monitor After initialize the tool bar will show the plate state and the connection status The plate in out is defined as green red The connection on off is defined as blue grey x Plate state OG Plate state 31L3C Connection e Connection Temperature monitor pa 8 78 Section F Working area The AgileCon allows you to define protocols and acquiring microplate data an exp experiment file will contain 2 information the Protocol and the Data The Protocol is to define parameters the data is to show experiment data B a Basic parameters well mapping Quantitative Cut off Ratio Inhibition OC Print options I Fie Experiment Setup Help A P Parameters Measurement type End Point D Filter wavelength nm Previous Current Maini 405 VW Referen
32. tall AgileCon Software 1 Start Windows 2 Close all unnecessary Windows programs 3 Place CD in the CD ROM drive 4 Copy the software from CD ROM to the Computer 5 Double click setup exe program 6 Click OK on the AgileCon setup menu Ep Welcome to the A eileCon installation program 2 metip cannot install syster files or update shared files if they are in v Before proceeding we recommend that you close any applications you may be TNL Goeres to install 8 Select Program group then click continue the AgileCon software will start installation 9 Click OK when the AgileCon finishes installation AgileCon Setup was completed successtully PE ne To start AgileCon 1 From Start menu gt Programs gt ACTGene gt AgileCon 2 Select the correct COM port for connection then click OK GY File Experment Setup Help Basic parameters Well mapping Quantitative Cut off Ratio Inhibition OC Print options Parameters Measurement type End Point D Previous Current Maint E Reference filter Filter wavelength mm Starting method Ze Immediate C Delay Plate motion Ze Continuous C Stepping i rincubation Use incubator Temperature p Ze Shaker Use shaker Measurement unit Unit None M Quantitative Cutoff M RatiosInhibition r ac COM port setting
33. the cutoff box and finished the configuration AMPR 750 will show the cutoff results according to the well mapping Depends on the conditions there will be max 5 symbols to interoperate the cutoff results C gt C gt C7 gt gt Ratio Inhibition results When clicking the Ratio Inhibition tab and finished configuring The AgileCon will show the sample OD value and standard OD value BO using Inhibition Ratio is represented in Blue Inhibition is represented in Red Over 200 the data will show Hi lower than 200 will show LO 1 2 3 10413 101 50 990 9652 59 92 0000 4007 34 27 0 00 DUU 9 60 10413 10190 99 0 9652 0992 0000 4007 34 27 UU 0 00 9 60 10413 101 50 99 0 9652 09952 0000 4007 34 27 UU 0 00 9 60 10043 9343 9200 44 00 35 39 10043 9343 92500 44 00 35 39 100 00 9343 92500 44 00 35 39 96 353 9543 92 00 44 00 35 39 96 53 9543 92 00 44 00 35 39 44 Q C results After clicking the QC calculation method and finished configuration the AccuMate will show the QC criteria Pass condition Result on the data sheet Results Calibration Ratiornhibition QC F H tic E C D E E H E L M ll Quality controls A Controls 3 control abs conc 4 PC 0 009 3 513 5 NE 0 073 110 217 D criteria 7 QC 1 1000 opt 1 000 NC 1500
34. tion 18 Bo well 19 Need quality control 20 Qc1 21 Qc3 22 Pass condition 25 24 plate layout 25 1 2 26 2i E 29 30 mom o gt Sample exp End point Continuous 450 Immediate No No Yes Curve on plate 5 5 Cubic polynomial Double threshold for gt 2 000 for lt 1 200 Inhibition C10 Yes 1 000 PC 1 000 NC 1 500 lt 1 200 0 500 lt 0 000 PC 1 000 NC 0 000 lt 1 000 QC QC1 AND QC3 3 4 5 6 7 8 9 10 11 12 SAMO1 1 SAM02 1 SAM03 1 SAM04 1 SAM05 1 SAM06 1 SAMO 7 1 SAMO8 1 SAM09 1 SAM10 1 SAM11 1 SAM12 1 SAMO1 2 SAM02 2 SAMO3 2 SAM042 SAM05 2 SAM06 2 SAMO07 2 SAMO8 2 SAMO9 2 SAM10 2 SAM11 2 SAM12 2 SAMO1 3 SAMO02 3 SAMO03 3 SAM04 3 SAMO5 3 SAM06 3 SAMO 7 3 SAMO8 3 SAMO09 3 SAM10 3 SAM11 3 SAM12 3 POS01 1 STDO1 1 STDO2 1 STDO3 1 STDO4 1 STDOS 1 POS01 2 STD01 2 TDO2 2 STDO3 2 STDO42 STDO5 2 4 Calibration When Quantitative is checked in the protocol calibration curve will be displayed according to the setting parameter Results Calibration Da B E D Ratiofinhibition J kK a a E L M N O 25 29 33 37 38 39 EA 41 E 43 KN 45 23 24 26 et 28 30 AL 32 34 35 36 46 ah E ZS A AL DE Calibrators col c02 c03 c04 c05 Name STDO1 TDO2 TDO3 TD04 TDO5 Meas 0 159 0 095 0 118 0 120 0 165
35. up Help OGOL 4 e Basic parameters ell mapping Quantitative Cut off Ratio Inhibition OC Print options Cutoff settings Cutoff method Single If result threshold then Threshold 0 e Positivel f Double f Calculation Ratio Inhibition The AgileCon will take a standard BO and other samples to calculate the Ratio Inhibition factor 1 Make sure the check mark is clicked on the Ratio Inhibition in Basis Parameter 2 Click the Cutoff tap in Ratio Inhibition 3 Define the desired parameter Si File Experiment Setup Help gt S F m le Ratio Inhibition e Ratio BBO C Inhibition 100 B BD Definition of HU s16 Q C The AgileCon can make Q C rules for experiment to determine the results 1 Make sure the check mark is clicked on the QC in Basis Parameter 2 Click the Cutoff tap in QC 3 Define the desired parameter Si File Experiment Setup Help op S ml s ELG General equation L aPC bMC c H L a h C H w OCI NIA si PC 1 MEA 1 5 sii Bes W DCH 0 5 Ek PC 1 MEA fi ait QG Pass condition CH QC true then PASS I l GO AND Qa 17 Print options Users can define the Name User and Printing selections for the experiments on the AgileCon To define print options 1 Click the Print options on the Basic parameters 2 Enter the desired field for print options 5i File Experiment Setup Help gt CESE Basi
36. ut of the experiment Raw abs matrix The OD value from the AMPR 750 Blanked abs matrix The OD value minus the blank Delta abs matrix The delta OD curve Calibration The calibration value Kinetics Kinetics value for the experiment Cutoff matrix Cutoff report 10 Ratio Inhibition matrix Ratio Inhibition report 11 Q C validation QC report 12 Data list Show the data list Matrix show as well mapping configuration 40 PE ne Interpreting the results The AgileCon will generate the results after every experiment in the data tab on the right Users can use to result Calibration Data Cutoff Ratio Inhibition Q C Kinetic to view their experiment results Result Click on the result tab to see the parameter setup plate layout Raw OD and Con Matrix of the experiment If check Quantitative cutoff Ratio Inhibition QC calculation method the criteria of the calculation method will also show on this page Results Calibration Data Cut off Ratio Inhibition Q C Kinetic Report B PS D E F G H I J K L M N Protocol parameters Experiment file path Measurement type Measurement mode Main_1 filter nm Starting method Need shake Need incubate W 100 OM GT 9 DI 1 Need quantitative 10 Quant method hl Quant standards number 12 Quant replicates number 13 curve fit method 14 cutoff method 15 Upper threshold label 16 Lower threshold label 17 Need ratio inhibi
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