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AmphaSoft User Guide

1. Buffer Results Flush Read Figure 6 AmphaSoft GUI with data plot 3 3 Work space and work list When starting a measurement the software prompts you with a pop up window Fig 7 for creating a workspace to save the data You can change the name in this window and confirm it with Enter When selecting an already existing name the system asks you for overwriting or changing Measurement data should be saved locally at the hard disk of the laptop or PC Don save the acquired date during measurement at a server or external drive connected to the laptop this will provide error messages and data loss You can also create a new workspace or load an existing workspace before starting your measurements with the menu bar Fig 7 right To select a generated workspace push the button marked with and pick your workspace in the list The system asks you for saving when loading an existing workspace you want to further analyze then click No D 1 D ip oct p sa sereme Sense sS G ferment it Figure 7 Creating or loading a workspace When you have created a workspace you can generate a work list by selecting at the bottom of settings Fig 8 A new window appears to plan and document your experiments Fig 8 right All your settings names and comments will be summarized in the report for documentation AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e
2. Comment Open File Location Change Icon Advanced LC oe JL ee Jl Ae J Figure 42 AmphaSoft gt Properties Target line with Service Mode Start now AmphaSoft in Online modus Go to Menu gt Help gt About and press the Update button as shown in the figure 43 About AmphaSoft AmphaSoft SW Version 1 1 0 FW Version 083 2 hasvs ore n NOTICE READ THIS END USER LICENSE AGREEMENT CAREFULLY BEFORE INSTALLING ACCESSING COPYING Op USING THE LICENSED PRODUCT ACCOMPANYING THIS AGREEMENT CLICKING ON THE I ACCEPT BUTTON BELOW OR IN ANY OTHER WAY INSTALLING ACCESSING COPYING OR USING THE LICENSED PRODUCT CREATES A LEGALLY IENFORCEABLE CONTRACT AND CONSTITUTES ACCEPTANCE OF ALL TERMS AND CONDITIONS OF THIS AGREEMENT Amphasys AG Technopark Lucerne CH 6039 Root D4 Switzerland 41 41 541 91 20 info amphasys com Figure 43 Start Update of Ampha Z30 firmware On the file section dialog specify either to show bin or bit files using the File name drop down menu and switch to the directory that contains the AmphaZ30 firmware file you downloaded from Amphasys Fig 44 Select the firmware file and press the Open button AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 31 AmphaSoft SW User Guide Q Select FW Look in B Desktop sl euer Ed3 les a Libra
3. InstallShield 3 Start AmphaSoft installation AmphaSoft 1 2 2 msi with double click and confirm any administrator or security rights When the installation is finished the AmphaSoft shortcut is created at the desktop Fig 26 A p FA ATTDHSSOIT Figure 26 AmphaSoft icon on the desktop 4 Start AmphaSoft by clicking the right mouse button and selecting gt Run as administrator Fig 27 When the software starts a picture is visible during the process Fig 28 left With advanced process a window pops up for selection of user modus use gt Offline and Continue Fig 28 right AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 24 Troubleshoot compatibility Open file location Wy Run as administrator Scan with Microsoft Security Essentials HP File Sanitizer Shred Pin to Taskbar Pin to Start Menu Restore previous versions Send to Cut Copy Create shortcut Delete Rename Properties Figure 27 Start AmphaSoft as administrator Continue Cancel Starting Figure 28 Starting AmphaSoft left and selection of Offline mode right AmphaSoft SW User Guide 5 When the AmphaSoft main GUI is open select in the menu gt Tools gt Chip gram your Ampha Z30 fluidic setup appropriate to your chip sizes Fig 29 Check for the AmphaSoft software version in Menu gt Help g
4. The expert window Fig 12 comprises 5 function modules Signal Actions Control Gain and Log Signal 1 5 IS ampi V 1 5 o i 2 0 2 6 8 10 12 time s 40 Control Gain Actions Level 0 1 Modulation lt gt 3 0 72Vpp CS Up 30 6 Amplifier lt gt e Viviy El El e Demodulation gt 2 4 Load EI Rinse Chip Detection d Pulse test Chip Test Trig e l 25 11 14 11 17 39 357 Info Demod scale 2 x4 25 11 14 11 17 39 341 Info Vrms measurement inactive 25 11 14 11 17 39 341 Info DC test signal disabled 25 11 14 11 17 39 341 Info AC filter on Figure 12 Expert Window details e Signal Display of data during the measurement or in test modus see Actions You can zoom in and out the amplitude at the y axis of the plot by clicking at the upper or lower number Fig 12 Changes Zoom of the x axis influence the measurement and recording of data When using this function change it back afterwards By zooming the x axis Fig 13 y you AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 12 AmphaSoft SW User Guide aise LOntrot Level 0 1 Pum y P 206 Vivixy Load Rinse 25 11 14 11 27 49 977 Info Measurement sens electrode on 25 11 14 11 27 49 977 Info Measure
5. AmphaSoft SOFTWARE USER GUIDE RELEASE 1 2 4 Workspace View Tools Help R CK m Settings Sample UI 1 Name 1 buffer id AF1 ei 0 9 0 8 Chip id 000000 kd 07 0 6 Freq 0 5 MHz 1 os 0 5 2 12 04 3 4 0 3 02 Stop Conditions SS ul min sec H Cells 0 ot 0 2 0 3 0 4 0 5 0 6 Results Concentration celis mi Flush y ee liste zu ud Ampitude 0 5MHz AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com AmphaSoft SW User Guide Content Bled ee ele BE 3 ACN r iaeMBone T 3 3 Software structure and parts 3 o H E 4 22 Mea IERI EE mmm 4 3 3 Work space and work Wer 8 S VOIDS RETE 11 33 FIC UU CUE REOR 11 36 do quee RU 12 3 Plot with zoom functions and population gang 18 CE e EM een E 20 3 9 Use of FCS files in EA G Joole nennen nnne nnns nnn nnn 21 4 Technical support updates and warranty n nsennoeneennoeneennsrrnerrnrrrenrsrerrrennennrrnnennrrnrererrenee 22 4 1 Software License and dongle cccccccccsececeeceeeeeeceeceeeeeeeseeesseeeeseeeessaeesseeeeseneesseeeesees 22 4 2 Software Installation and Update 00nnannnannnannnennnnnnnnnnnnnennnrnnnrennrrnrrrnrrererrrnrrrerrrenerene 22 a 2T Amps
6. For technical or software support contact Amphasys AG Technopark Lucerne Platz 4 CH 6039 Root D4 Switzerland Phone 41 41 541 91 20 Email info amphasys com AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 33 AmphaSoft SW User Guide Index AmphaSoft Installation 23 AmphaSoft Main GUI 4 ACION ne eae 5 7 ES OL OMIA et 7 Fess 7 SE 4 7 EE 4 AmphaSoft Update 22 BONO IO oou eee anne ee Rene non Rone none noe eat UU LM oe Ter 6 9 WD AG WEEN 6 9 CAD OUA eeri 17 Chip ECSU m 17 Cont check box eese nnns 10 Data file structure esee 11 Done check box eene 10 Seine ele e EE 12 A ege 13 Ampltter esses 15 CIOS LECT e MM 16 CAD Eege 16 Be Lait de ee ee 13 Nat Ile 14 DEMOOUIALON BEE 15 Cr M 15 Ec 13 BE AO RR 14 Boo X 16 Modulation eee 15 xul MO 13 mila E 14 RG i el EE 12 Signal overflow 15 Hair mA 13 uoc c 11 21 gl ETC E rena nautae sinas 30 ER CARPERE 7 10 Wu UNI 6 9 Biet 3 22 Notes for measurement 10 Offline MOGUS eseou 3 Online Ale e UC 3 Parameters f
7. Printer Printer Copy 1 Default Programs 4 Unspecified 1 PowerPoint 2013 Help and Support ep o Create shortcut AY Troubleshoot Unknown Device Properties Sg Unknown Device Category Unknown 1 Status Needs troubleshooting Shut down mm Figure 32 left Start menu open Devices and Printers right Control Panel Devices and Printers Unspecified Device gt Properties 8 The window for Unknown Device Properties appears select the tab gt Hardware and click on gt Properties right at the bottom Fig 33 m P b Control Panel Hardware and Sound Devices and Printers nte Adda device Add a printer Troubleshoot Bluetooth Mouse DISK CD M557 Hardware lii Unknown Device Brother Device Functions Microsoft XPS PaperPortImage PDF Complete Send To Document Writer Printer OneNote 2013 Name fa Unknown device Printer 4 Unspecified Device Function Summary Manufacturer Unknown Location Location D Port_ 0002 Hub_ 0002 Device status The drivers for this device are not installed Code 28 Figure 33 Unknown Device gt Properties Tab gt Hardware 9 A second window pops up select in the tab gt General Change settings left at the bottom Fig 34 left The window changes possibly you have to confirm administrator rights select then gt Update Driver Fig 34 right AMPHASYS AG TECHNOPARK Lucerne Pl
8. Trig displays the signals of a possible measurement based on the given parameters It allows you to test for the real signal and to verify changes of the parameters and settings before starting the real measurement To stop it press Bl Stop B ends the selected action Do not press this button during active measurement only stop your measurement in the main window Control 1 manipulation of the recording and control of pump and valves Level This is the trigger value set to record the signals When using in Actions All or Trig it is visible in the Signal area as a red dotted line above and below x axis The level should be adjusted to a value about double the signal noise and lower than the signals you want to record pos neg drop down function Selects the direction of the signal curve from the signal for recording It can be verified by using the Actions function The normal signal with X positive or Y negative is seen in Fig 12 Pump Here you can manually control the pump with check uncheck a given value between 0 and 400rpm and the direction of rotation The rotation is counter clockwise from sample tube to the chip with unchecked valves normally used and clockwise from water bottle to the sample tube or chip You have to use it with the right valve positions Be carefully with the direction and speed it is dependent from the fluidic set and chip and can mix your sample or pollute your water To adjust the speed of the pump
9. 2MHz XR 54 21k Sta ul min sec Cells Figure 16 Calibration left normal chip right calibration with warning due to unusual values AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 16 AmphaSoft SW User Guide Chip Test detailed explanations Process Two calibration resistors in the electronics of the device are first measured They are connected to the electric circuit paths of the two measurement electrode pairs and their measurement checks the integrity of the electric line to the chip Then the impedance of the liquid between the two electrode pairs is measured first between the sensor electrode pair which is also called measurement electrode pair and is situated closer to the chip inlet and then between the reference electrode pair These measurements can be compared to ideal measurements of a good chip with PBS buffer as liquid Large inconsistencies result in a warning Calibration generates for every frequency four outputs 1 Reference Angle of the device with PR PM P the reference angle of the instrument 2 Liquid impedance R P the average impedances of the two electrode pairs 3 Reference electrode values RR PR the reference electrode pair impedances 4 Measurement electrode values RM PM the measurement electrode pair impedances These outputs are important to check chip quality and chip comparabil
10. Copy 1 4 Unspecified 1 Figure 39 left window Update Driver Software 1 Ampha Z30 close the window right Control Panel window device identification Ampha Z30 15 Start AmphaSoft in Online modus Fig 40 left and select in the menu gt Help gt About a Online SW Version 1 1 0 FW Version 082 0 Offline NOTICE READ THIS END USER LICENSE AGREEMENT Continue CAREFULLY BEFORE INSTALLING ACCESSING COPY Lee OR USING THE LICENSED PRODUCT ACCOMPANYING NW A AGREEMENT CLICKING ON THE I ACCEPT BUTTON B OR IN ANY OTHER WAY INSTALLING ACCESSING CC 4 n p Amphasys AG Technopark Lucerne CH 6039 Root D4 Switzerland 41 41 541 91 20 Figure 40 left 1 AmphaSoft Online modus right menu gt Help gt About recognition of instrument by showing a firmware version 4 3 Firmware Update You will be informed per Email if there is any firmware update of the Ampha Z30 instrument necessary You can check the firmware version in the AmphaSoft software in the menu with gt Help gt About Fig 41 The information you get in a pop up window Wonspace Wew Toots Help SCI D Usor Guide Amphaz 30 User Matua settings a rine a e s z on l AmphaSoft SW Version D FW Version 083 2 Amphasys AG End User License Agreement NOTICE READ THIS END USER LICENSE AGREEMENT CAREFULLY BEFORE INSTALLING ACCESSING COPYING OR USING THE LICENSED PRODUCT ACCOMPANYING THIS AGREEMEN
11. Figure 19 Creating reports 00 00 39 The report files are saved automatically in the loaded workspace When the htm report is generated an additional folder is saved in the workspace with single pictures of all your plots in png format Fig 20 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 20 AmphaSoft SW User Guide Date modified Type 8 7 2014 7 50 AN ile folder Share with 7 Slide show Burn New folder A 8 7 2014 7 39 AM File folder Name Date 8 7 20147 45AM_ File folder m AmphasysLogo 3 24 2014 11 01 PM 8 7 2014 7 50 AN File folder MS 1 1 8 7 2014 7 56 AM s S_1 2 L wp72 report res 8 7 2014 7 56 AM MS 2 1 wp72 report csv 8 7 2014 7 56 AN CSV File 2 KB 8i S22 wp72 report 8 7 2014 7 56 AV HTML Document 6 KB MS 3 1 Me S 3 2 Figure 20 Files and folders generated from reporting function 3 9 Use of FCS files in FACS tools From each measurement a file in FCS3 format is generated and saved in a special folder in the workspace Fig 20 The files include all information to load and to analyze with a typical Flow Cytometry Data Analysis Software such as FlowJo version 7 6 5 ideal higher versions still with bugs Flowing Software Infinicyte etc When loading your data in the tools use all parameters in linear scale The parameters you can select for plotting etc are e for each meas
12. Update Driver Software Unknown Device 11 Select with bottom gt Browse the folder C AmphaSoft AmphaZ30Driver Fig 36 ate Dri er Software Unknown Device Browse for driver software on your computer Search for driver software in this location C AmphaSoft AmphaZ30Driver V Include subfolders 4 jV Computer 4 amp Local Disk C 4h AmphaSoft d AmphaZ30Driver gt Let me pick from a list of device drivers on my computer di images This list will show installed driver software compatible with the device and all driver Jj Usb software in the same category as the device b AN Brother Folder AmphaZ30Driver Figure 36 Update Driver Software Unknown Device Browse for Folder C AmphaSoft AmphaZ30Driver AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 28 AmphaSoft SW User Guide 12 The selection should look like Fig 37 also check gt Include subfolders and go to gt Next Browse for driver software on your computer Search for driver software in this location Include subfolders gt Let me pick from a list of device drivers on my computer This list will show installed driver software compatible with the device and all driver software in the same category as the device EL Figure 37 Update Driver Software Unknown Device driver location 13 A Windows Security message asks fo
13. OIL AOC E 22 4 2 2 AmphaSoft Installatnon 23 Ao cd i i E 30 Zo Bn e VV CUNY E 33 4 5 Extended Warrantv 33 AO SUDO ERR mE 33 lj qe T 34 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com AmphaSoft SW User Guide 1 Introduction This User Guide provides detailed information about the software used with Ampha Z30 impedance flow cytometer for controlling the instrument data acquisition and data analysis which runs on a Windows based laptop PC Fig 1 Data should be saved locally at the hard disk at the laptop PC The use of the software AmphaSoft needs a license which will be provided with a USB dongle This stick has always to be plugged in the PC or laptop during measurement or analysis For more details see section 2 For matters regarding the Ampha Z30 instrument accessories chips etc please refer to the Ampha Z3 User Manual or go to http www amphasys com community Ampha Z39 Figure 1 The Ampha Z30 Impedance Flow Cytometer System 2 License and Dongle The license for using AmphaSoft will be provided with an USB dongle which always has to be plugged in the PC or laptop during online and offline use USB dongles are available with full license For trial or test licenses contact Amphasys The dongle contains the license file which should never be d
14. P 64 5 sel M ee ea m E Reference Electrode Values Chip Bk ADDE E E z po 3 ap F 0 50MHz RR 9089 08kOhm PR 10 9 ec ZP Linh Le a F 200MHz RR 5521 1b5kOhm PR 44 1 S Id AC14010 id AC140108 F 6 00MHz R 55 68kOhm P 44 F 6 00MHz RR 2536 32kOhm PR 62 8 F 12 00MHz R 56 86kOhm P 4 8 5 F 12 00MHz RR 1355 39kOhm PR 64 7 Reference Electrode Values R Measurement Electrode Values F 0 550MHz RR 53 10kOhm PR 2 10 F 0 50MHz RM 11627 07kOhm PM 25 2 F 200MHz RR 5650kOhm PR A D 0 2 0 4 F 200MHz RM 6390 93kOhm PM 57 3 Freq 0 5 MHz 1 x m Freq 0 5 MHz 1 F 6 00MHz RM 2684 56kOhm PM App r L Gab F 12 00MHz RR 59 84kOhm PR 3 1 Control Gain d F 12 00MHz RM 1457 65kOhm PM 64 5 1 0 5 Level 0 1 v Modulation 2 Measurement Electrode Values P 2 2 o I N F 0 50MHz RM 47 79kOhm PM 7 8 FUP sog Amplifier 3 6 F 2 00MHz RM 50 48kOhm PM 4 9 Fe H 4 1 2 F 6 00MHz RM 52 73kOhm PM 5 2 wa DI EI 9 Demodutaion f F 12 00MHz RM 5431kOhm PM 6 6 ep n B sensor liquid contact 1 l Load Rinse L oK Log FH f electrodes out of specs system still may work Fa quer iiri E i rri rni ferons c JA verify liquid conductivity check wetness of the sensor contacts check Info Liquid Impedance F 12MHz XR 45975 sensor state Stop Conditions Infa _Liauid_ Imnedance F
15. any time by clicking on Stop Rinsing It is recommended however to perform at least one complete rinsing cycle before starting with the first measurement e Experimental setup Before starting a measurement details for the sample used buffer and chip can be entered in the upper area 1 o Sample Id Name Depending on the number of samples you plan to analyze see section 3 3 AmphaSoft automatically enumerates your Sample Id from S 1 to S n with n standing for the last sample of the series You can change this Id according to your own discretion if required o Buffer Id Select from the pull down menu Fig 5A one of the proposed buffer types you plan to use for the measurement If it is not listed add it to your list or replace a present one as needed by selecting o Chip Id Likewise select or add the correct Chip Id in format X00000 Fig 5B Using not this format will give you an error Fig 5C The Chip ld is visible on the chip Entering the right Chip Id is a must and selects the necessary parameters for the fluidic processes The software detects the chip type and gives an error message for AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 5 AmphaSoft SW User Guide Q Amphasoft a discrepancy Selecting the Chip Id will help tracking the usage of the chip for the various measurements and providing comparability
16. for optimal measurement check the signals with the Trig function in Actions For ideal speed most signals are in the range of 1 to 4 x10 seconds Fig 12 2 in the given time frame Avoid speeds below 1 x10 seconds which could cause false counts Optimal speed for each tubing set and AmphaChip are in the AmphaZ30 user manual in section 3 1 4 table 3 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland phone 41 41 541 91 20 info amphasys com e www amphasys com 13 AmphasSoft SW User Guide Valve The valves are in standard position unchecked Be carefully when changing this function X Y Selects the real X or imaginary Y part of the signal for recording It can be verified by using the Actions function The normal signal with X positive or Y negative is shown in Fig 12 Load This function affects the measurement function in the main window With the unchecked box it is possible to measure a sample which is already loaded in the instrument by a previous measurement or a manually loaded sample Rinse This action influences the initial Rinsing process when starting the instrument and software The first Rinsing has to be performed before measurement Only uncheck this box if you have to restart the software for special reasons and if you have a sample already loaded More details about the fluidic and the manual use of the pump and valves are explained in the Ampha Z30 user manual Def
17. phone 41 41 541 91 20 info amphasys com www amphasys com Mana wms Figure 8 Creating a work list In the work list the default parameters for one measurement are displayed To add more samples select for adding each time one sample or with a number and to add more samples With you can delete samples again if they were not done Enter your Sample Names edit the Buffer ld Chip Id frequencies parameters and stop criteria if necessary Fig 9 Change parameters Level Mod Amp Dem Pump only if you Know your values otherwise use expert window section 3 6 and confirm The pump speed for measurement is set with the Chip Id any changes can influence your results All adds and changed values beside chip parameters can also be adapted and changed in the normal AmphaSoft window in section lt settings gt and will be saved AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com AmphaSoft SW User Guide Amp Dem Pump 2 MH MHz DA bal Dat e DR bpm ul min sec Cells Flash Cant Freq 1 Freq 2 Freg 3 Freg 4 Mean Frog Level Mod tampnis ld Samplo Mame Buffer Id Chip kd Hiz MH 5 1 xyz AFI Anaon KE 2 i2 os 0 1 3 H 1 aon E fap AO given number of measurements P Freq Freq 3 Eng d X Era LEw Med Dem Fu Sample id Sample Mame Buffer Id Chip id MHz MHz MHz ec iE a nau D ul mie sec Celi
18. G TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 10 AmphasSoft SW User Guide fDoneodstands for a finished measurement It can by not selected by the user It is automatically checked after a finished measurement Samples with a checked Done box cannot be deleted To switch between samples during measurement or analysis use the button at the lower left corner in main AmphaSoft GUI 3 4 Template For routine or repeating measurements using the same Chip lds settings and names we recommend to save your generated or measured work list as a template Also generated gates are saved with this template and applied to your new measurements Use in the menu bar Workspace Save as template and give a new name Load this template for your next measurements and save it if necessary again with a new name and you can directly go on with your measurement without additional changes Generate also such templates if you want to change any extra settings or parameters 3 5 File structure The creation of a workspace and measurement of samples generates the file structure shown in Fig 10 The data of each measured sample is stored in an own folder that includes 5 files The files amph clr meas and isf are specific for AmphaSoft The file csv can be loaded as text file in Excel like programs for special analysis The folder FCS includes all generate
19. T CLICKING ON THE ACCEPT BUTTON BELOW OR IN ANY OTHER WAY INSTALLING ACCESSING COPYING OR USING THE LICENSED PRODUCT CREATES A LEGALLY ENFORCEABLE CONTRACT AND CONSTITUTES ACCEPTANCE OF ALL TERMS AND CONDITIONS OF THIS AGREEMENT tepee Zebcteg NI Sat Mei rene saart rang Amphasys AG Technopark Lucerne CH 6039 Root D4 Switzerland 41 41 541 91 20 info amphasys com Figure 41 Check for the firmware version of Ampha Z30 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 30 AmphaSoft SW User Guide To update the firmware start the laptop and switch on the Ampha Z30 You have to start AmphaSoft in service mode To do this click with right mouse button on the AmphaSoft logo on your desktop Select gt Properties and gt Shortcut Fig 42 Add in the field gt Target at the end G S the complete line should be C AmphaSoft CellRecorderStarter exe C AmphaSoft CellRecorder exe AmphaSoft G S Confirm with gt Apply and gt OK could be that there pops up a window with administrator rights 1 confirm that too e AmphaSoft Properties Security Details _ i __ Previous Versions _ SSt ee ee General Shortcut Compatibility AmphaSoft Target type Application Target location Ampha Soft Start in C AmphaSoft Shortcut key None Run Normal window
20. aain m Figure 14 Expert Window specials e Gain The three sliders Modulation Amplifier and Demodulation allow the change of the instrument sensitivity Default settings are 3 6 2 x4 Changes should be always controlled with the Actions function Modulation influences the electric power applied on the chip Only change the Modulation with expertise for example for increasing the sensitivity while using low conductivity buffers or analyzing small cells It should not be marked red as shown in Fig 14 which means that the applied power is too high In case of signal overflow using default values the chip might be blocked gt check flow rate or out of specifications gt perform chip test Please note that higher voltages can accelerate the degradation of the chip electrodes and reduce the life time of the chip Amplifier modifies and enhances the signal to noise ratio It can be reduced when signals are too high and cut or when it is marked red signal overflow To strengthen the signal you can increase the value but it should not turn red Possibly you have to adjust the Level Demodulation changes the signal and noise value in the same range by enhancing or decreasing Reduce it when it changes to red or the signals look strange Control and adjust the Level if necessary Pulse test offers the possibility to test the instrument electronics and chip connection It can be used with a normal chip and running c
21. ata obtained from a former experiment Fig 2C Measurement data should be saved locally at the hard disk of the laptop or PC Don save the acquired date during measurement at a server or external drive connected to the laptop this will provide error messages and data loss A BIET Mode Online C Offline AESi Cancel Figure 2 A Instrument B Software shortcut icon on desktop C Selection of online or offline user ken Measurement The AmphaSoft Main GUI is divided in three sections Fig 4 1 2 3 Settings Measurement parameters are selected here Information about sample buffer and chip are entered as well as the measurement frequencies and the measurement stop criteria All these settings are stored for each measurement and supplied in the report files Results This section provides on the one hand a graphical output of the measured cells in terms of a scatter plot in which the signal Amplitude y axis is plotted against the signal Phase x axis The dots are shown as a density analysis in which highest cell density has red the lowest dark blue color On the other hand also cell concentration cell counts and the rejection rate of the analyzed cells are shown Actions This section supplies the commands for the measurement and rinsing procedures and indicates the status of these actions with the status bar below Before starting any process or measurement you have to select the t
22. atz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 2 AmphaSoft SW User Guide Device type Other devices Manufacturer Unknown Location Location 0 Port amp 0002 Hub 0002 Device type Other devices Manufacturer Unknown Location Location 0 Port_ 0002 Hub_ 0002 Device status Derce aint The drivers for this device are not installed Code 28 The drivers for this device are not installed Code 28 There is no driver selected for the device information set or There is no driver selected forthe device information set or Be element To find a driver for this device click Update Driver To find a driver for this device click Update Driver Update Driver Figure 34 left second window of Unknown Device Properties use tab gt General gt Change settings right 1 Unknown Device Properties select gt Update Drivere 10 In a further window Update Driver Software Unknown Device you can select the driver software with gt Browse my computer for driver software Fig 35 How do you want to search for driver software zb Search automatically for updated driver software Windows will search your computer and the Internet for the latest driver software for your device unless you ve disabled this feature in your device installation settings Browse my computer for driver software Locate and install driver software manually s ES Figure 35
23. ault settings and parameters are Level 0 1 X positive or Y negative Pump unchecked counter clockwise Pump speed dependent from selected tubing set and chip type set S 20rpm A chip set M 40rom B chip 80rpm C chip set L 60rpm D chip 300rpm E chip Valves unchecked Load and Rinse function checked Signal plot i Amplitude at 10 V and time frame 15x10 s Gain 3 Modulation 6 Amplifier 2 Demodulation These settings are loaded by starting AmphaSoft Any changes are saved to the user account on the laptop and are loaded with the AmphaSoft to any user To prevent any trouble work with default and special application templates AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland phone 41 41 541 91 20 info amphasys com e www amphasys com 14 AmphaSoft SW User Guide CG j Q AmphaSoft Expert Window o gt el se Signal 1 I ampl V 2 0 2 6 8 10 12 time s a Control Gain Actions Level 0 1 7 Modulation gt 4 1 4Vpp E PumP olg Amplifier ds Vivixy e Demodulation H gt 2 x4 oad El Rinse Chip Detection Pulse test Y t p 25 11 14 11 30 07 790 Warning ADC overflow Please reduce modulation gain z 25 11 14 11 30 06 868 Warning ADC overflow Please reduce modulation gain 25 11 14 11 30 06 368 Warning ADC overflow Please reduce modulation gain 25 11 14 11 30 05 940 Warnina ADC overflow Please reduce modulation
24. d fcs files in FCS3 format which can be loaded and analyzed with a suitable software tool section 3 9 When reports were generated Section 3 8 the documentation files csv to open in Excel or similar tools and or html to open in a browser and an additional folder is added with pictures of all plots in png format Sic 5 Users Amphasys E AmphaSoft gt wp72 gt ze Nindo Amot Intel mig eril Pro Pro SWT Users A Arm kt A 4 D k KL J k k KL 1 D k i k Li Figure 10 File and folder structure AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 11 Te AmphaSoft SW User Guide 3 6 Expert window The Expert Window permits application development with the opportunity to change and optimize parameters It is selected with gt Tools gt Expert Window opening an extra window which is always on top Fig 11 You can minimize or close it any time you have done your changes or tests Changes in the settings or parameters are saved to the user account on the laptop and are loaded with the AmphaSoft to any user To prevent any trouble work with default and special application templates Bam Workspace View Tools Help Dewls SJ Expert Window Settin gs Tubing Instrument Cleaning Sample id _1 Name id A00000 Frea 0 5 MHz 11 Figure 11 Starting Expert Window
25. d values pops up Fig 16 If the test impedances are far away from normal e g defect chip air or water in the fluidics an additional warning window pops up in the end of the test Fig 16 right Chip Detection This function is normally active Only uncheck if you know what you do e g for special cleaning processes otherwise you flood the instrument and cause electronic damage e log displays all actions and steps running at the AmphaZ30 instrument during measurement It is also saved as a text file in the AmphaSoft folder Workspace View Tools Help Workspace View Tools Heln El Calibration Results See Settings Settings Reference Angle of Device Sample Sample F 0 50MHz PR 27 8 PM 28 0 P 27 9 id P s id s F 200MHz PR 8 0 PM 88 P 84 x 4 l AmphasSoft Expert Window F 600MHz PR 04 PM 18 P 0 7 LX N Name E Calibration Results 0000 al die F 1200MHz PR 7 PM 8T P 5 7 x Reference Angle of Device U F 0 50MHz PR 27 8 PM 28 0 P 27 9 GE kee F 200MHz PR 8 0 PM Z P 84 od E ids DE Se aoe pam F 2 00MHz R 5967 82kOhm P 50 2 id PCB F 6 00MHz PR 0 4 PM 1 8 P 0 7 5 PCB F 6 00MHz R 2611 98kOhm P 64 7 F 1200MHz PR 7 7 PM 87 P 57 F 1200MHz R 1405 41kOhm
26. eleted and the dongle should not be used for any other purpose 3 Software structure and parts AmphaSoft Software runs in Offline or Online mode with the option to perform routine experiments in User level with default values or in Expert Window section 2 5 for application development and the opportunity to change parameters Offline Online Data Analysis Data Acquisition Planning of Experiments Planning of Experiments Reporting Data Analysis Reporting AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com AmphaSoft SW User Guide Many functions or buttons are briefly described with a mouse over or hotspot when moving with the mouse cursor to the respective position Further Information about the Ampha Z30 instrument is supplied in the separate Ampha Z30 User Manual 3 1 3 2 Start Instrument Check liquid bottles The front bottle contains normally deionized water the bottle at the side is for waste liquid and should be preferably empty at the beginning of a test series Turn on the instrument the switch is located right at the backside Turn on the laptop PC Make sure that the USB cable is correctly connected to your device Start the software rAmphaSoft by double clicking at the icon on the desktop Fig 2B Select your user mode rOnlinedif you plan a measurement or foffline if you want to load and analyze d
27. er Guide Boos M B Se Due to electronic settings in the E o J instrument the displayed data look like t To shift the X axis you have to select gt View gt Phase Correction and adjust the value in the field P H Works View Tools Help When you want to adjust the Zoom or Phase correction to all samples you have to be in the corresponding active mode and by clicking with the right mouse button on the plot you can select gt Apply to all an rauen Start Raung AmphasSoft gives you the opportunity to select for subpopulations with percentage numbers in a line or polygon gating function Both gating function cannot by applied to a sample Gates and subpopulations are listed and displayed in the reports The gating function is selected with gt View gt Line or gt Polygon gating Fig 18 The gate can be removed or applied to all samples in the corresponding active mode with the right mouse button Figure 18 Line and Polygon gating 8 A en s Line gating p E The line gating is a partition in 4 quadrants By adjusting the cross to the top or bottom of the plot you can use it as bifurcated gate in a histogram AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 19 AmphasSoft SW User Guide B Polygon gating ee a The polygon gate allows a selection of a Se subpopulation with ow
28. ity A good quality chip should lie between the following optimal values at 0 5 MHz measured with 1xPBS or 0 9 NaCl solution see table Chips with bigger channels gt 50um have normally smaller values than these with small channels 7 um Measurements with low conductivity buffers show higher values Impedance R at 0 5MHz 1x PBS Impedance R at 0 5MHz 0 5x PBS AmphaCalib Asymmetry between electrodes all chip types Table Classification for x with various Sizes E d at 0 5 MHz with 1xPBS and 0 5xPBS AmphaCalib AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland phone 41 41 541 91 20 e info amphasys com e www amphasys com 17 AmphaSoft SW User Guide An acceptable chip can still be used as far as its measurement results are not analyzed together with results from other chips In general chips with similar liquid impedances give similar cell impedances Therefore results from chips with similar liquid impedances might be interchangeable A broken chip should not be used any more as it will not separate cell types nicely or is leaky in the instrument When you have high values check first if there is flow through your chip if there is the liquid you want to measure in the channel and no water or no air bubbles One cause of high impedances what makes chips get into the acceptable or broken category is the formation of a biofilm on the electrodes This can be removed by 10 min incubati
29. l be stopped as soon as one of the entered criteria is achieved AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com AmphaSoft SW User Guide Make sure that your sample volume is in agreement with your settings Living these fields blank requires your manual intervention to stop the measurement Use only realistic values e Flush check box 1 fFlushostands for a short rinsing cycle of the whole tubing system including the chip using system liquid usually deionized water It is part of a measurement process and cannot be performed separately It starts right after the measurement has been stopped manually or automatically and prepares the instrument for the next measurement It removes from in the lines most cells of the just terminated measurement but is not a thorough cleaning After this short flush cycle the device is ready for loading the next sample Check this box if you plan serial analyses for which extensive rinsing of the system in between the different samples is not required Note the Flush cycle pumps water into the sample tube Thus if you check this box be aware that the rest of your sample will be diluted If you still need the residual sample do not check this box or exchange sample tube immediately after the measurement was stopped e By selecting at the bottom of 1 you open a work list to plan and document yo
30. ment ref electrode on 25 11 14 11 27 49 977 25 11 14 1127 4 change the time frame in which the signals are collected during measurement The default time frame is 15 x 10 s Fig 12 When changing the x axis to a shorter time frame Fig 13 left i time frame 5 x 10 s signals from slow running particles are not counted or rejected due to imperfect signals When zooming out the x axis Fig 13 right time frame 30 x 10 s signal processing becomes worse and inaccurate 1 5 3 time s lime s Modulation 3 0 72Vpp Modulation 3 0 72Vpp Amplifier 6 Amplifier 6 Demodulation 2 x4 Demodulation 2 x4 Chip Detection v Chip Detection v Pulse test v g 25 11 14 11 27 49 977 Info Measurement sens electrode 25 11 14 11 27 49 977 Info Measurement ref electrode on Info Demodulator input 6 gt gain transimpedance gain 4 743 Ohm 25 11 14 11 27 49 977 Info Demodulator input 6 gt gain transimpedance gain 4 743 Ohm Modulator nutnut 3 gt voltane 0 7234 Von Aodulator eutnut 3 gt valtana N 7234 Vnn Figure 13 Expert Window change of time frame in signal plot Actions Test modus of measurement with Stop Bi All and Trig function X part is blue Y part is green All shows the continuous output of the instrument in the Signal area like an oscilloscope before data processing with selection When you want to check a sample for signals noise or any other tests activate this box To stop it press B
31. n naming To select a gate start at the upper left side and circle the population with left mouse button clicks To close the gate make a GES double mouse click Now you can name o the gate WC 0 _ With the right mouse button gates can be sc removed or applied to all samples Polygon gates can be moved when you want to adjust it to other measurements signals new workspaces etc Activate the polygon gating function in the menu bar and use gt Ctrl together with A nq Ka rLofor moving up left down or right 3 8 Report AmphasSoft contains a reporting function for detailed documentation of your measurements including plots and all gating information After finishing your analysis the report can be generated in the menu with gt Workspace gt Report Fig 19 You have the option to generate a csv report delimiter separated values or a htm report The csv file can be open in Excel like tools and it contains all information settings and gating details but no plots The htm files can be started in a browser installed on your PC and contains all information and details including the plots Workspace View Tools H Settings Name Sample eport t E 140708uc report res Id Name A FCS Strg Q 1 kSsa e kS2 Buffer Si KAS LSA Buffer AC140108 Freq 0 5 MHz 4 minsec 1 bi 350 Accepted Rejected Del Start Rinsing Ready NEN IAN UE
32. of the measurements Workspace View Tools Help ug d AS nho Settings Sample Id Name Buffer D00000 Freq 0 5 MHz 1 1 2 3 4 Stop Conditions ul min sec Cells Results Accepted Concentration cells ml Rejected X ejecte Start Measurement Figure 4 AmphaSoft GUI A B C Bufera e BI X chip id ei x Q Chip Id Form eji ams Sw 1 Wi 1 A00000 2 ap m Chip Id must begin with a capital letter al ms A E followed by 5 digits e g 12345 Came o cancel Ok Figure 5 Pull down menu for Buffer Id A for Chip Id B selection and format of Chip Id C O Selection of measurement frequencies Choose the desired measurement frequencies in the middle part 1 you can select 1 2 or 4 frequencies For the first frequency a low value 0 5 MHz 0 5 to 2 MHz is recommended to have optimal signals for recording the measurement For higher frequencies you have to check the signal with the Expert window 3 6 The other frequencies you can apply in the range of 0 1 MHz up to 30 MHz Note For using 4 frequencies apply first the fourth and then the third frequency e Inthe lower part of section 1 you can select among three different stop criteria for your measurement volume time or number of measured cells If you enter two or all three criteria the measurement wil
33. on number Fig 46 right Previous Versions Compatibility AmphaSoft Target type Application Target location Ampha Soft xe C AmphaSoft CellRecorder exe Ampha Sofi Start in C Ampha Soft About AmphaSoft Shortcut key None Run Normal window Comment SW Version 1 2 2 ion 082 0 Open File Location Change Icon Advanced FW Version License ID BACDEFGHIJKLMNOPQ s AG End User License Agreement INOTICE READ THIS END USER LICENSE AGREEMI CAREFULLY BEFORE INSTALLING ACCESSING C IOR USING THE LICENSED PRODUCT ACCOMPANY 4 Amphasys AG Technopark Lucerne CH 6039 Root D4 Switzerland 41 41 541 91 20 Figure 46 left AmphaSoft properties change of Shortcut Target to normal user modus right Check Ampha Z30 firmware version AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 32 AmphaSoft SW User Guide 4 4 Limited Warranty Limited warranty period is one 1 year from first operation of the Product by Buyer It covers software and firmware and defects in material and workmanship under normal use and does not apply to ordinary wear and tear For detailed information refer to the warranty information at www amphasys com 4 5 Extended Warranty Extended warranties are available Visit www amphasys com for information 4 6 Support
34. on with AmphaClean solution followed by rinsing with water 3 Plot with zoom functions and population gating After each measurement a density plot of the collected data is generated with adjunct histograms for x axis at the top and for y axis at the right side Fig 17 The progress of the data calculation after measurement to display the final plot is shown in the lower bar It can take some seconds when a high number of events were collected The zoom and gating functions of the plot can be used with Online and Offline AmphaSoft To optimal visualize the data you can change the display with the corresponding icon in the toolbar or in the menu gt View gt Zoom and using right mouse button to select a frame or by adjusting the values in the right upper corner details in Fig 17 To change between View mode you have to deselect the active mode by clicking on it in the menu bar or escape Figure 17 Density plot and adjunct histograms with zoom functions A To adjust X Phase and Y Amplitude use in the toolbar the icon or in the menu gt Zoom mode or type in numbers in corresponding field for the axis in the right Upper corner Separate the numbers with Se a space tutter By using lt R gt or Athe selection is reset Accepted Ready AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 18 AmphaSoft SW Us
35. onductive buffer with low to normal speed in the fluidics Checking the box and monitoring the signal in Actions All shows an electronic pulse generated on the chip Fig 15 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 19 AmphaSoft SW User Guide E 1 0 5 REES LEE a L a a EE EE Ee D T em ti epe ep ger aepo qo atqne T s er prp e s 0 5 uk 15 2L 0 0 2 0 4 0 6 0 8 1 12 1 4 e time s um Control Gain Actions 4 Level 0 1 Modulation gt 3 0 72Vpp a Sep Ampie Se Vivixy PIP o Demodulation af gt 1 x2 ead i Rinse Chip Detection V Pulse test V Chip Test Trig L p LG 25 11 14 11 56 39 692 Info Measurement sens electrode on 25 11 14 11 56 39 689 Info Measurement ref electrode on 25 11 14 11 56 39 687 Info Demodulator input 6 gt gain transimpedance gain 4 743 Ohm be 25 11 14 11 56 39 624A Info Modulator outout 3 gt valtane 0 7234 Vnn Figure 15 Pulse function Chip Test This function determines the impedance of the liquid that runs through the chip and the instrument With normal chips the pump has to be active with low or measurement speed 1 to 100rpm At the end of the test process takes about 20 seconds a window with the measure
36. or measurement 9 10 Planing and adding experiments 7 zi 7 18 Cris 18 EME GAUN E 19 POIVGON GAUN ersen 19 tee P 18 PUMP SDCCG ME 9 PRG ON GE 8 11 20 zi 20 Hmm 20 FUINSING PIOCESS 5 SANE acteurs ernie nnana 5 Sample Name ccscccceeeeceeeeceeeeeeeseeeeseseneesees 9 Stop criteria for measurement 7 9 i i 5 11 TUDING SO aso RE 5 JE Ile UE 3 22 Work list EE 7 9 NV OIMS DACG e 8 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland phone 41 41 541 91 20 info amphasys com e www amphasys com 34
37. or rights or account at the laptop you like to install the software Create a folder on your desktop or somewhere else Download the files from the Amphasys website and save them in the folder Fig 24 Ca Al gt AmphaSoft Installation hae Organize v jl Install Y Share with v New folder H 9 gd reegt Name 3 Date modified Type Size EE Desktop m3 AMD64 all rootsupd 2f61a03102b7aededed0362ab4fd023d2016f5f0 18 08 2014 09 51 Application 448 KB Ip Downloads 9 AmphaSoft 1 1 0 28 07 2014 14 32 Windows Installer Package 25 572 KB Z OneDrive _ AmphaZ30 Firmware 0820 140624 0904 bit 18 08 2014 09 53 BIT File 1 452 KB i Recent Places E McCRInstaller 18 08 2014 09 51 Application 177310 KB i Libraries Ee Documents a Music i Pictures Figure 24 Download folder for AmphaSoft software installation 1 Start installation with file AMD64 all rootsupd by double click and confirm it if any windows for installation or security rights appear 2 Continue installation with MCRInstaller by double click and confirming The installation process needs some time and requests for your input Fig 25A C AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 23 AmphaSoft SW User Guide Le Tel s v 4 Search AmphaSoft Instaliation Dp b AmphaSoft Installation Nd ee Or Open Share
38. r installation check gt Always trust Software from Mlriamec Motion AGoand select Install Fig 38 Would you like to install this device software Name Amphasys Universal Serial Bus controller IT Publisher Triamec Motion AG j aN trust software from Triamec Motion AG Le You should only install driver software from publishers you trust How can I decide which device software is safe to install Device Function Summary Manufacturer Unknown Location Location D Port 0002 Hub 30002 Device status The drivers for this device are not installed Code 28 o J ewe J Apply Figure 38 Windows Security message and installation of driver 14 After finishing installation process close the window Update Driver Software Ampha Z30 Fig 39 left Check if in the Control Panel window the Unspecified device is identified as Ampha Z30 Fig 39 right AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland phone 41 41 541 91 20 info amphasys com e www amphasys com 29 Add a device Add a pnnter Bluetooth Mouse DISK CD M557 Windows has successfully updated your driver software 4 Printers and Faxes 9 Windows has finished installing the driver software for this device 7 7 7 7 7 7 Brother PC FAX ax HP ePrint Microsoft XPS PaperPort Image PDF Complete Send To MFC J6910DW MFC 36910DW v22 Document Writer Pnnter OneNote 2013 Printer Printer
39. ries Homegroup Recent Places System Folder Ki System Folder sochosp KR Computer Desktop System Folder PS System Folder Be E e Network E zoe MM zx KE System Folder B PERS raries J 1 41 MB i A eS Computer a Network Gage AmphaZ30 Firmware 0820 140624 0904 bit D Files of type AmphaSoft FW C bit M Cancel Figure 44 File of Ampha Z30 firmware Do not turn off the instrument until the software prompts you that the firmware update process has completed successfully Fig 45 SS SSS About AmphaSoft Le m ies AmphaSoft Lei SI Jee OR USING THE LICENSED PRODUCT ACCOMPANYING THIS AGREEMENT CLICKING ON THE Il ACCEPT BUTTON BELOW OR IN ANY OTHER WAY INSTALLING ACCESSING COPYING OR USING THE LICENSED PRODUCT CREATES A LEGALLY ENFORCEABLE CONTRACT AND CONSTITUTES ACCEPTANCE OF ALL TERMS AND CONDITIONS OF THIS AGREEMENT 7 Firmware update successtul Powercycle instrument Amphasys AG Technopark Lucerne CH 6039 Root D4 Switzerland 41 41 541 91 20 info amphasys com Figure 45 Update of Ampha Z30 firmware Now close the AmphaSoft program and hit the On Off switch at the back of the instrument Wait a few seconds and switch the instrument back on Remove the G S switch from the AmphaSoft shortcut properties window Fig 46 left and start the AmphaSoft program by double clicking on the shortcut Go to Help gt About You should now see the new firmware versi
40. s Flush Coni m xy 00000 2 d sa ve A00000 a tg SA xyz ADDOD0 5 5 ET Kl EN 5_6 xyz Ann 2 3 2 1 d 1 1 S3 xy 1 2 x IEDDDD3 i s 3 3 H 1 1 1 1 1 Freq 1 Freq 2 Freq 3 Freq 4 Maas Freg Level Mod Amp Dem Pump Sample id Sample Name Buffer id Chip ld MHz MHz MHz MHz 1 4 WM pD 5 e m a ipm ul mimsec Cells Flush Con hate Done SI ayz AF jA00000 5 E 6 12 0 5 ER 3 6 i 300 E 2 2 ayz ADOO00 m as 2 6 12 os Di 3 1 300 iF 5 3 xyz AF ADODO 45 2 6 12 0 5 8 1 i 6 1 300 y SA ayz ru t 0 5 2 e 12 Da 0 1 3 B 1 300 B 53 xyz AF1 AfD x 0 5 8 12 Da KR 3 1 300 F 5 6 xyz AFA ADODOD 5 2 5 12 KE Di 3 5 1 300 7 i NS Figure 9 Complete a work list add samples add Sample Name change Buffer Id or Chip Id format X00000 change frequencies add stop criteria un check Flush Cont make notes change parameters Level Mod Amp Dem Pump Caution e Flush check box explained in section 3 2 e Cont check box Cont ostands for automatic measurement continuation It can be selected in a work list for routine measurements After a sample is measured plotted and flushed the measurement process moves to the next sample according to the settings in the work list and the loading step is directly started The system asks you for the new sample to continue e Done check box AMPHASYS A
41. t About You will get a pop up Window with details to the software Fig 30 Close the program without saving the workspace Amplitude 2MHz M d 2 Phase 2 0 MHz 50 Concentration cells ml 7342420 WEM sU 3X 4 J J s ooo ee EE 00 00 15 Figure 29 AmphaSoft main GUI selection of Fluidic Setup and close program AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 25 AmphaSoft SW User Guide won TM Workspace View Tools Help oda A LD Settings AmphaSoft SW Version H FW Version 082 0 License 1D BACDEFGHIJKLMNOPQ Acmohasys AG End User License Aessen TE NOTICE READ THIS END USER LICENSE AGREEM CAREFULLY BEFORE INSTALLING ACCESSING Cx OR USING THE LICENSED PROOUCT ACCOMPANY Amphasys AG Technopark Leceme CH 5039 Root D4 Swtzertand 41 41 541 91 20 Results Concentration celis mi Stan Measurement Figure 30 Check AmphaSoft software version 6 Connect the instrument with the laptop with the USB Mini cable and switch on the instrument 1 a pop up window at the desktop right at the bottom appears click on it Fig 31 left You will get the information in an extra window that the device driver was not installed Fig 31 right Close the window ini a ed Device driver software was not successfully installed Please cons
42. the dongle and start the software again I E in USB license to proceed C Starting Start Error Figure 22 Error messages without USB dongle for AmphaSoft license ID 4 2 Software Installation and Update 4 2 1 AmphaSoft Update You will be informed per Email if there is any software update of AmphaSoft available To check your AmphaSoft software version start the software and check in the menu with gt Help gt About Fig 23 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 22 haSoft SW User Guide Figure 23 AmphaSoft software version During Installation AmphaSoft should be closed and not running in Online or Offline mode For installation you should have administrator rights for your laptop or PC For updating download the new software version from http Wwww amphasys com community to the laptop Start the AmphaSoft x y z msi file follow the instructions and the new software will be installed at the laptop After finishing you can use AmphaSoft with the normal icon and check the software version Fig 23 4 2 2 AmphaSoft Installation When setting up a new laptop for measurement with Ampha Z30 the following installation process has to be completed AmphaSoft software and associated drivers and installation tools can be downloaded with a user account from http www amphasys com community You should have administrat
43. ubing set you will measure This can be chosen with gt Menu gt Tools gt Program Fig 3 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com AmphaSoft SW User Guide d p Workspace View Tools Help OHsea ixac Expert Window Tubing d Large Settings Instrumen t Cleanin g Medium Sample Small Id 1 Name 1 Cells Flush Eh ird eG O dde Figure 3 AmphaSoft Main GUI 1 Tubing set selection Starting AmphaSoft will load default settings user level which can usually be applied for routine measurements demanding only few changes To keep these default values save the initial workspace as a template see section 3 4 and load this template for routine measurements without any extra changes in the parameters After adding Chip Ids and any new parameters you will use for various measurement days we recommend to save the workspace as a separate template section 3 4 and to load this template for your new measurements The following explanations of AmphaSoft GUI using 1 2 3 are referred to Figure 4 e Before starting the system for measurement execute the rinsing process by clicking on You will be prompted to check the statRinsing presence of a chip of an empty tube on the sample holder and the liquid level of water and waste bottles After starting this process you can stop it
44. ult with your device manufacturer for assistance getting this device installed USB 2 0 Hub off Ready to use Unidentified Device 3 No driver found t Installing device driver software X What can I do if my device did not install properly Click here for status 10 18 Close Figure 31 left AmphaSoft properties change of Shortcut Target right 1 Check AmphaSoft software version a Y vo Be fg RE Boda RA 7 Open in the gt Start menu of your laptop gt Devices and Printers Fig 32 left The Control Panel window opens and shows to your laptop connected devices The bottom line gt Unspecified displays an Unknown Device Open with right mouse button gt Properties Fig 32 right AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 26 AmphaSoft SW User Guide ke a Control Panel Hardware and Sound Devices and Printers Search Devices and Printers Add a device Add a printer Troubleshoot Documents P 2 Pictures C e Paint Bluetooth Mouse DISK CD Music MSS7 l Getting Started Recent Items a Printers and Faxes 9 Connect to a Projector Computer BE ho eee ai ai m Sticky Notes MFC J6910DW MFC J6910DW v22 Document Writer Printer OneNote 2013 R Snipping Tool Devices and Printers Brother Brother Brother PC FAX HP ePrint Microsoft XPS PaperPortImage PDF Complete Send To
45. ur experiments Details are explained in 3 3 Work space e With lt gt you select between samples during measurement and analysis 1 e The bottom line informs you about actions to do or during the run 1 e Results and plot are displayed in 2 Actions are displayed in 3 e When you want to measure push the button Start Measurement you will be ask to save the data to a workspace more details in 3 3 and to provide a chip and a sample always continue with a mouse click or Enter e The continuation of the process is shown with the long bar in 3 and in the bottom line of 1 e During measurement there is first the sample with high pump speed loaded than the pump speed is adjusted and cells are detected e The button is changed to Stop measurement and you can stop the measurement any time or it will be stopped by your given criteria e In 2 the plot of measured cells will be updated regularly At the bottom there are the number of accepted cells the percentage of rejected cells and the cell concentration per ml displayed and regularly updated until stop e After stop of measurement the data are calculated and the plot is updated to a density plot with red yellow dots showing high number and blue dots for low number of events Fig 6 AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland phone 41 41 541 91 20 info amphasys com e www amphasys com AmphaSoft SW User Guide Sample
46. ured frequency X and Y as real and imaginary part of the signal Rand P as amplitude and phase of the signal calculated values as opacity from X Y Rand P e transit time time which a particle needs between both measurement electrodes e time total measurement time AMPHASYS AG TECHNOPARK Lucerne Platz 4 CH 6039 Root D4 Switzerland e phone 41 41 541 91 20 info amphasys com www amphasys com 21 AmphaSoft SW User Guide 4 Technical support updates and warranty 4 1 Software License and dongle With the purchase of the Ampha Z30 instrument you get an USB dongle with a license file for using Amphasoft The license ID you can check with the AmphaSoft software in the menu with gt Help gt About Fig 21 A dongle is a small security device that plugs into your computer It works like a mobile serial number for AmphaSoft which allow running AmphaSoft on several computers but just one at a time About AmphaSoft jm AmphaSoft SW Version 1 2 2 FW Version 082 0 License ID BACDEFGHIJKLMNOPQ NOTICE READ THIS END USER LICENSE AGREEMI H CAREFULLY BEFORE INSTALLING ACCESSING C OR USING THE LICENSED PRODUCT ACCOMPANY 4 a aed oot ST aT m Em Amphasys AG Technopark Lucerne CH 6039 Root D4 Switzerland 41 41 541 91 20 Figure 21 AmphaSoft software version license ID If the USB dongle is not plugged in the computer you will get an error message Fig 22 Press gt Esc plug in
47. with v New folder WY Favorites Ner RE Desktop m3 AMD64 all rootsupd_2f61a0a102b7 ae Jg Downloads PI AmphaSoft 1 1 0 Z OneDrive _ AmphaZ30_Firmware_0820_140624 09 Recent Places E McRinstaller Date modified requirements Ss DN e M TLABIDI Compiler Runtime 7 14 requires that the following requirements be installed on your computer prior to installing this application Click OK to begin installing these i53 Libraries 3 Documents m Music i Pictures B Videos Status Requirement Pending VCREDIST X85 Ba Computer amp Local Disk C cx HP TOOLS D ex Removable Disk E cx HP RECOVERY F Network Compiler Runtime The InstallShield R Wizard will install MATLAB R Compiler Runtime 7 14 on your computer To continue dick Next MATLAB and Simulink are registered trademarks of The MathWorks Inc Please see www mathworks com trademarks for a list of other trademarks owned by The MathWorks Inc Other product or brand names are trademarks or registered tradmarks of their respective owners WARNING This program is protected by copyright law and international treaties Copyright 1984 2010 The MathWorks Inc The MathWorks Figure 25 Process during MCRinstaller setting up Click Next to install to this folder or click Change to install to a different folder CA Install MATLAB R Compiler Runtime 7 14 to C Program Files x86 MATLAB MATLAB Compiler Runtime

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