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Stimulus Generator Manual STG 4004 and STG 4008

1. AAAARANAR WYSIWYG Stimulus protocol display T CLL a GO o0 Oo o00 Ot 0 001 Oro Oc OO Otro Ot OO Doi Ot 004 OCcO Ot 005 jg Oc Cee OO DO aau ida ija DO Cc COE Coles DO og 0 006 t hi EE zzz A aja 2 amp k e on m ee i oa je os r cs roe r o ree set F Sie T Sea I Smet T Smet T Smet I Sime F eady 3 2 Stimulus Generator 4000 Series Manual Setting Up and Connecting the STG Note You can use a USB hub for connecting the STG to the computer for example if you have no free USB port or if you need to extend the USB cable Do not use a hub in Streaming mode Please note that if you want to use the STG in Streaming mode you cannot connect a second device that sends or receives continuous data streams for example a web cam or USB speakers to the same USB port because the STG needs a broad bandwidth for the data transmission This does not matter if you want to use the Download mode via MC Stimulus II or custom program only Provide a power supply and a computer with USB port in the immediate vicinity of the installation site Make sure the STG is switched off before you connect it to the power supply Place the STG on a stable surface where the air can circulate freely and the STG is not exposed to direct sunlight Connect the external power supply to the STG Connect the external power supply to the power outlet Connect the U
2. STG 4008 Rear Panel 4 0 0 0 0 0 0 0 0 DIG ro uiis SYNC OUT TRIGGER IN jn 0000 OOOO 1e VOC USB Driver Installation The stimulus generator is a plug and play device The driver is automatically installed together with the MC Stimulus Il program The Windows operating system detects a new hardware when the stimulus generator is connected to the computer if the program has not been installed beforehand Simply cancel the driver installation and proceed with the installation of the MC Stimulus Il program Important Please make sure that you have full control over your computer as an administrator Otherwise it is possible that the installed hardware does not work properly If there are any problems and you need to install or update the driver manually please see the MC Stimulus Il help for more information 3 4 Stimulus Generator 4000 Series Manual Installing the Software System requirements Software One of the following Microsoft Windows operating systems is required Windows 7 Vista or Windows XP English and German versions supported Hardware USB port 2 0 High Speed Installing the software Please check the system requirements before you install the software MCS cannot guarantee that the software works properly if these requirements are not fulfilled Important Please make sure that you have full control over your computer as an administrator Otherwise is possible that the i
3. Shift gis Absolute value 1n TOI 2 00 00 00 000 00 00 00 200 Dior an 00 400 00 00 00 600 00 00 00 800 00 00 07 000 00 00 01 200 t hh mm ss zzz 29 Stimulus Generator 4000 Series Manual The absolute values of 200 and 200 are both 200 This absolute value of 200 is modified by the shift factor of 50 resulting in 250 The negative value is now 250 and the positive value is 200 If you had chosen the option Shift instead the computed result would have been 200 50 250 All zero values are not changed Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 vr Puse voie unt time une Fs unt tme unt vote unit me unt Row repeat Group repeat eo o o C m m d 3 frectenguer 0 m a sme fm S jrecangie B recae zjredenuer m DJ bo Ln J md 3 ft is 1 1 Dor ad ad dao Do ad ad 2060 Do an ad 4060 Dr ad ad eo Do adr ad so 00 00 01 000 Bo adr 1 200 t hh mm zs zzz Autocreating entries You can use the Autofill feature to set up advanced time or voltage current increments You define the Start and End value and either the Step Size Start Stop Step Size or the number of Steps Start Stop of Steps Alternatively you can enter the Step Size and the Number of Steps Start Step Size of Steps According to these specifications rows are inserted and the column
4. 77 maximum output current 74 99 measuring output with an oscilloscope 77 OSCITOSCODG aiii ober raso uM ae ud 74 output mode 19 69 74 LesolUtIOPL siete iei Terba repre RE 20 Cursor 45 Cut LOWS aiani a i pd aoi o 54 Shortcut Kay ui mise aaea 54 D Data ASCII file format 36 download sueta edet en e eR 49 SIVO isset ia vedette nM Ad ede Du ere 55 Digital blanking signal 57 LED EE PRE AAEE EE E bm 8 58 OUTPUT nian eee ee 8 57 DO rnaen 8 Display Auto SYNC output 31 STOD DE eese iuuat cua Ear an ues 45 selecting channels 45 Stimulus Generator 4000 Series Manual SEELIITQS ziii iesu ced eames Goiamaales ZOOM EPI Downloading patch oO orit ie eiie erum SDhOPECUT KEY iu oic vs codo cR SUmulus THes ione eto cies accces Driver installation eee eee nnne Duration Edit Electrode CAPACITIVE behavior End Error Messages Extended Multi File Mode F Faraday cage CORDec HTIO sea tacui FERE C Uu DERES 70 File Batcn MOG CG nisse EE Mta r d importing ASCII files Format ASCITO eoin LID Oe XIS ao a G GND outputs aDOLU E usas or e teme Ope elg Eb connecting arseron a Ede 70 Ground Group Group Repeat eese enne
5. 4 frectenguer i0 mv 40 us 400 mv 5b mV s S frectenguer 100 mV 50 ue 40 mw s0 s mW s B recangier 100 mwe us 400 mvjso us mv s i um e p cen ce i em E e T p en ce E i T p cen e p p cn pem Dor ad ad dao Dr an ad a1 t hh mm ss zzz In the same way you can set up a stimulus with voltage current increments Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel amp Channel 7 Channel 8 a a I I KT UE RET E CYOGETH T frectanguer Elf mv 20 s 0 mv s ms 2 frectanguer ERN nv so Qe 0 mv us mv 05 3 rectencuer DM nv 40 s X mv 0 e mv us 4 frectenguer EMM mv eo s aoo mv us mV s S jrecenguer Milf nv 40 s aoo mv so s mv us B_frectenguer MEM mv 40 s 00 mv us mV s 00 00 00 ao 00 00 00 a t hh mm ss zzz Shifting absolute time or voltage values With this option the signs of the selected values are ignored Zeros are never changed that is the option Keep Zero cannot be deselected Under Type of Increment select Absolute Value Enter the desired shift factor Channel 1 Channel 2 Channel 3 Channel 4 ChannelS Peia zug ER TE rz gt unes SERT Ei Increment selected values rectangular 200 mw 20 ms Zr CE A i Tux Cancel a 3 frectanguer 0 ms m M Type of Increment 5 ES a C Count frecur am i Keep Zero scena ae DJ
6. multichannel systems Stimulus Generator Manual STG 4004 and STG 4008 Information in this document is subject to change without notice No part of this document may be reproduced or transmitted without the express written permission of Multi Channel Systems MCS GmbH While every precaution has been taken in the preparation of this document the publisher and the author assume no responsibility for errors or omissions or for damages resulting from the use of information contained in this document or from the use of programs and source code that may accompany it In no event shall the publisher and the author be liable for any loss of profit or any other commercial damage caused or alleged to have been caused directly or indirectly by this document 2015 Multi Channel Systems MCS GmbH All rights reserved Printed 04 06 2013 Multi Channel Systems MCS GmbH AspenhaustraBe 21 72770 Reutlingen Germany Fon 49 71 21 90 925 0 Fax 49 71 21 90 92 5 11 info multichannelsystems com www multichannelsystems com Microsoft and Windows are registered trademarks of Microsoft Corporation Products that are referred to in this document may be either trademarks and or registered trademarks of their respective holders and should be noted as such The publisher and the author make no claim to these trademark 1 1 1 2 1 3 2 1 2 2 2 3 3 1 3 2 3 3 3 4 4 1 4 2 5 1 5 2 5 3 5 4 5 5 5 6 9 5 8 6 1 6
7. 3500 30060 4q Values 3500 enu enu I J I channels of I I format J channel I pulse eae Os eae Os eae Os ee Os I channel I pulse ls Os I channel I pulse Os Us Os Os I channel I pulse Os Os ls Os am Os I I values onoo 4000007 2000007 icad value e agdgagauqr OO000T values OOO00T ZOOO00T values 4000007 4000007 ongoa t ime t imeT t ime t ime 41 Stimulus Generator 4000 Series Manual After loading the file into the MC Stimulus II worksheet you can edit and download the file as usual Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 Sync 1 Pulse vatue Unt time unit vaue uni time Unit vatue un ume unit Row rep Group r s recente so me o ms mv us mv s 2 ramp Jo mv 899 mv 200 ms 71 ss Xo m mo m m a frectanguer m s mv e fm Ss r n0 2000 3000 2000 r 2000 3000 2000 S000 1000 1000 3000 r n 2000 3000 2000 m T a ann 00 00 00 000 00 00 00 500 00 00 01 0000 00 00 01 5900 t hh mm zs zzz L1 uz Ls Lad Format type 6 Note This file format is used by MC Rack ASCII export This format type is available only in the current version of the import filter This is a basic format with two columns The absolute time in seconds and the absolute value of vo
8. Personnel still undergoing training may only work on the device under the supervision of an experienced person Guarantee and Liability The General conditions of sale and delivery of Multi Channel Systems MCS GmbH always apply The operator will receive these no later than on conclusion of the contract Multi Channel Systems MCS GmbH makes no guarantee as to the accuracy of any and all tests and data generated by the use of the device or the software It is up to the user to use good laboratory practice to establish the validity of his findings Guarantee and liability claims in the event of injury or material damage are excluded when they are the result of one of the following Improper use of the device Improper installation commissioning operation or maintenance of the device Operating the device when the safety and protective devices are defective and or inoperable Non observance of the instructions in the manual with regard to transport storage installation commissioning operation or maintenance of the device Unauthorized structural alterations to the device Unauthorized modifications to the system settings Inadequate monitoring of device components subject to wear Improperly executed and unauthorized repairs Unauthorized opening of the device or its components Catastrophic events due to the effect of foreign bodies or acts of God Stimulus Generator 4000 Series Manual Important Safety Advice Warning Make su
9. otherwise they count like 1 high It is recommended to build a connector All channels are assigned to trigger 1 Please make sure that you do not use any other trigger input Each time when trigger 1 is activated the program applies that one of the stimulus pattern which currently is selected by DO to D7 64 Synchronizing Events Assignment of the digital inputs D7 to DO Settings C emm Hewm 1 0 0000000 CTERgRSTG 4DODWI 8164000 Greuphepeshstm ooo mox 4 booooio CHERgRSTG ROUUWK STG4DUD Overviewsim L8 oo001000 CHEirgfiSTG ADODMC STG4DUO Grouphepeshsim 0010000 CBRSTG ODDC STO4DOD GroupRepesbsim ae m Don 00000 2 Birgitvs Tes 3000 WC S TG Overview stm Fa oad Save Show all Patterns Decimal Binary Pattern D7 D6 D5 m3 ok sa Goococcecea Smecoococcecae Seesacoceca cococococotrt coccococceocu Serres e oc oa ocn Seecocounucese Seoococoocounces The user is asked to compose his own stimulation pattern files Please do not create too big stimulus files because all files are completely downloaded and stored in the memory of the STG If the files contain too much data and you apply a lot of files you will get an error message on download The advantage of this mode is to have all stimulus files directly in the memory of the STG and thus there is no time lag when starting a new stimulus pattern on each trigger Tr
10. 0 B frectenguer o ZJrectencuier Dor ad ad dao Bo ad ad 2060 Do ad ad 4060 Dur ad ad eo 00 00 00 200 t hh mm ss zzz 00 00 07 000 Do adr 01 200 Incrementing columns This feature is very convenient for setting up long and complex stimuli without entering each value manually You can enter and edit multiple voltage values or time lengths You can keep all zero values unchanged with the option Keep Zero Apart from an absolute shift or increment factor you can modify values by the percentage as well Shifting time or voltage current values You enter a value which is added arithmetically to all values in the selected column For example if you enter 50 50 is added to all values Click any Value or Time column header to select a column The selected column is highlighted in black 26 2 3 4 Programming Stimulus Protocols Right click and click Increment Selection Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel amp Channel 7 Channel 8 Sy Pulse value Unit time vale Unit time Unit value Unit time Unit Row rep Group r change Time Unit Invert selection 4 EN rectangular rectangular rectangular ni TT unn mu 5 r HE AM EI An Dor ad ad dao 00 00 00 0001 t hh mm ss zzz Under Type of Increment select Shift Enter the desired value Channel channel2 Channels ETETTERETTISTSTS xp
11. 60 Auto Download stimulus intensity level 32 Autocalibration 11 GISQDIG iret devant PME 11 Autocreating SUM aeaa PE impo rta 25 Autofill 25 Autorepeat pulses or complete pulse protocols 22 B Back panel 8 Batch MEN oerion act odes ta ned deas uud 53 io E 67 Batch Mode 67 Biphasic pulse 20 91 Blanking amplifier ei oe etii 57 autocreating blanking pulses 31 SION OT iaee mte 57 BNC connection to oscilloscope 77 COMNEGCIONS isis e ts De eae 8 C Cables ShieldTo sis i bid cns 70 74 twisted Dall sosisini a e 70 Capacitive behavior of electrodes 91 Cascade 53 Change All stimulus intensity level 32 Change Time Unit 25 Channels CONN ECUNI Mene NE 8 displaying c a odes heh bees tua gto se ruind 45 downloading sess 49 editing estt PEE 19 Clear file assignment in Multi File mode 63 Compiler warnings 56 Connecting the STG to an amplifier 8 the STG to an oscilloscope 77 Contact IDTOPFDUSUOF esie secteur eo torent 101 Continuous SUmulatiOh suse one ne i dorin 22 Copy FOWL idis idi eia s dus 54 SHIOFCCUT KO ssim e dass Ma E vet edad 54 Count 25 Creating Autosync pulses eeeossssssss 31 SCION oido donc ls Doa ADR DR 19 Current capacitive behavior of electrodes 91 electrode impedance
12. A logic state of 1 means 3 3 V and a logic state of 0 means 0 V Please note that the digital output is about 15 ps faster than the analog output This small offset is generally sufficient to make sure that the Sync Out signal precedes the stimulation which is important for synchronizing events You can program the output signal of the Sync Out channels in the same way as the analog output signals but due to the nature of a digital signal only values of 0 and 1 are allowed A Sync Out signal is generated each time when the STG is started The output signal can be used for triggering external devices for example the data acquisition with the MC Card of the ME System and the MEA System Please note that if you use it for triggering the MC Card you should use output signals that have a duration of at least 200 ps Otherwise it can happen that the MC Card ignores trigger events All four Sync Out outputs are active when the STG is started by pressing the Start Stop button or by using the software command Start on the STG menu If you are using the Trigger In inputs for starting the STG each of the four Sync Out channels is controlled by a separate Trigger In channel Sync Out 1 is active when Trigger In 1 is triggered Sync Out 2 is active when Trigger In 2 is active and so on The assignment of Sync Out channels to Trigger In inputs can be customized please see Trigger Settings If you use the Repeat feature in the Trigger Settings dialo
13. Independent Channel Download Operating multiple MEA Amplifiers with one STG General Software Features Customizing the Main Window Menu Bar Toolbar Shortcut Keys File Menu Settings Menu Synchronizing Events Digital Output Signals Sync Out Triggering Stimulation Trigger In Trigger Settings Multi File Mode Extended Multi File Mode Batch Mode Analog Output Signals Output Modes Voltage Mode Current Mode Measuring Output Signals with an Oscilloscope Rise Time Comparison of Output Signals in Current Mode Capacitive Behavior of Stimulation Electrodes Troubleshooting About Troubleshooting Technical Support Error Messages Signal terminated when using Trigger In Strong Peak Artifacts Output Signal Does not Match Programmed Signal Signal is not Repeated Pulse Train Fails Internal Calibration Signals Appendix Technical Specifications STG4004 and STG4008 Pin Layout Digital O Contact Information Index 49 49 49 50 51 53 53 53 54 54 55 56 57 57 58 60 63 64 67 69 69 70 74 77 77 85 91 93 93 93 94 96 96 96 97 97 99 99 100 101 103 1 1 Introduction About this Manual This manual comprises all important information about the installation and operation of the stimulus generator STG4000 series and MC Stimulus II software It is assumed that you have already a basic understanding of technical and software terms No special skills are required to read th
14. UoN as desti estado hac os uuo 10 requirements eese eise eene 10 Start channels on trigger 60 files on trigger eeeeeeee 63 shortcut Key eb acta ie 54 7 f COMERS ree ee er ere n eres ei toss Prata 11 Starting poiht of ramp us saei va uon 20 STG COMM CCTING usce Up merde 8 WASHING ctr S 9 PTA IN MM 53 SARUN cna cto erededuissateso me tette adis 11 58 Stimuli ChCATIAG ET E cea 19 Stimulus Files ASCII Import without Header 36 downloading sssssss 49 triggering multiple files 63 Stimulus Intensity Level Ol OSTING sonis shuts pronti tnde md prod 32 Stop Batch Oei sia 67 channels on a trigger 60 files on trigger eeeese 63 SHOVLCUL KEY isis oct up ietn als 54 SIG u 54 StODDIDO STG osre 11 Support 93 Sweep LED Start Stop buttons 11 starting STG on trigger 11 Status JdmpDiz esci eapi nabiera 56 sweep COUNT wn Ane 60 CIO O CHING oie iu eir px Hebe icace 60 triggering sweeps eeeee 58 Switch circuit GIGOTGTTE o op Dice cas 58 Sync Out autocreating Sync Out pulses 31 CONMOCIONS E 8 digital TTL outputs 57 1 14 276 To oie REREN 60 System requirements 10 T Tile 53 Time axis TORO s od NER 45 change time units s
15. be consistent for all lines in a given ASCII file for example value time value time This error message indicates that there is a current voltage value or a time point missing in line 12 The ASCII file contains current voltage values outside the STG s range For example if the STG s output range is 8000 mV all values above 8000 mV are set to 8000 mV 95 12 4 12 5 12 6 Stimulus Generator 4000 Series Manual Signal terminated when using Trigger In If you use the Trigger In to start the STG and the stimulation protocol is terminated right after start check the Trigger Settings If the Use Trigger as Gate function is selected a protocol will only be active as long as the Trigger is HIGH If the Single Trigger function is selected the STG is armed and waits for an external trigger to execute the stimulus protocol one time only Switch to Stop Restart or Ignore Strong Peak Artifacts Strong discharges or other artefacts of the output voltage or current Possible causes One or both current outputs are open that is nothing is connected to them The stimulus generator tries to hold a stable current against the indefinite resistance of the open current output Therefore the stimulus generator will increase the output voltage on the open current outputs to its maximum which will result in a heavy load on the power supply This situation may cause the power supply to become unstable which may lead to artifacts on
16. but channel 3 in file A is still active As channel 3 from file A is assigned to trigger 1 the settings defined for on channel assigned to same trigger would be applicable Trigger 1 is active but channel 3 in file B is still active As channel 3 from file B is assigned to trigger 2 the settings defined for on channel assigned to other trigger would be applicable Extended Multi File Mode The Extended Multi File Mode is a very flexible mode for advanced users It can be used with stimulus generators of the 4000 series with a digital IN OUT connector only that means STG 4004 and STG 4008 It is possible to download up to nine different stimulus patterns in the default setting of the Extended Multi File Mode Selecting the option Show all Patterns the program supports 256 different patterns of stimulation You can download up to 256 stimulus patterns in advance and select among them You can use one of these stimulus patterns at a time of your choice with a digital signal Please read also chapter Multi File Mode This high number of different stimulus patterns are selected via the digital IN OUT connector Please see the pin layout of the 25 pin D Sub connector in the appendix Pin 1 to pin 8 of the connector refer to digital input DO to D7 Pin 9 to pin 16 refer to digital output D8 to D15 but they are not connected Pin 17 to pin 25 are grounded If you do not use all eight digital inputs you have to connect them to ground manually
17. of the programmed stimulation pattern the display will show 1 3 2 3 3 0 4 15 62 Synchronizing Events 10 4 Multi File Mode As an alternative to triggering separate channels in a single stimulus protocol you can also trigger up to four separate stimulus files in the Multi File mode This feature allows you to trigger different stimulus protocols on overlapping channels For example you can control stimulus pattern A by trigger 1 on channels 1 and 2 and stimulus pattern B by trigger 2 on channels 2 and 3 and so on The Multi File mode is indicated in the MC_Stimulus status control bar in the main window If you use the Multi File mode you cannot use the trigger inputs anymore for triggering single channels When you start the download all selected up to four stimulus files are downloaded onto the stimulus generator When Trigger In 1 is active the file assigned to Trigger 1 is started with all downloaded output channels when Trigger In 2 is active the file assigned to Trigger 2 is started and so on You can choose what action should be taken when a trigger event occurs during the time when the assigned file is still playing If the STG is started by the START STOP button on the front panel or the Start command of MC Stimulus II only the first top selected file this is usually the file assigned to trigger 1 in the Trigger Settings dialog box is started For downloading and starting multiple files without triggering please
18. retailer The highly qualified staff will be glad to help you Please inform your local retailer as well if other problems that are not mentioned in this documentation occur even if you have solved the problem on your own This helps other users and it helps MCS to optimize the instrument and the documentation Please pay attention to the safety and service information chapter Important Safety advice in the Manual Help Multi Channel Systems MCS GmbH has put all effort into making the product fully stable and reliable but like all high performance products it has to be handled with care Technical Support Contact your local retailer immediately if the cause of trouble remains unclear Please understand that information on your hardware and software configuration is necessary to analyze and finally solve the problem you encounter If you have any questions or if any problem occurs that is not mentioned in this document please contact your local retailer The highly qualified staff will be glad to help you Please keep information on the following at hand Description of the error the error message text or any other useful information and of the context in which the error occurred Try to remember all steps you had performed immediately before the error occurred The more information on the actual situation you can provide the easier it is to track the problem The serial number of the device You will find it on the device or cli
19. s er s CENE sI Tgger 7 e C G B E a C 3 O uj Fa Cancel In the preceding example trigger input 1 is used to trigger the output channels 1 3 and 6 This means each time a TTL pulse is applied to Trigger In 1 stimulation on channels 1 3 and 6 is started simultaneously The complete protocols of these three channels would be executed three times after the trigger event if continuous mode would be off However as continuous mode is activated trigger repeats are disabled and the protocols will be repeated indefinitely until a manual stop occurs Important Only channels assigned to the same trigger have the same time base Between channels assigned to different triggers time shifts in the order of the STGs time resolution 20 us may occur The Sync Out outputs are assigned to their default trigger inputs that is Sync Out 1 is active when trigger 1 is active and so on You can change this assignment with the option buttons for example assign multiple Sync Outs to the same trigger The Sweep status light LED Start Stop buttons indicates the trigger numbers that have been assigned to active channels Example If channels 1 and 3 are assigned to trigger 1 and trigger 1 is started either by the software controls the Start button on the stimulus generator s front panel or the trigger input 1 the first Sweep status lamp indicates the active state of channel 1 or 3 even if one of the two channels has a short
20. should always take the impedance of the stimulating electrodes into account when planning your stimulus protocol You should also keep in mind that most electrodes or amplifiers support only limited voltages You can increase the compliance voltage to 120 V by using the and outputs The output current even at the maximum output voltage of 150 V should not be dangerous to human health because the maximum output current in current mode is only 1 6 mA See illustration Doubled maximal output voltage A bipolar stimulation is recommended for extracellular recordings from microelectrode arrays MEAs if a very high output signal density in a defined area between two stimulating electrodes is needed The current that is delivered by the first electrode flows directly back to the second electrode so that the surrounding tissue is not affected See illustration Dipolar stimulation Screen shot from a standard oscilloscope triggered on the Sync Out pulse see Measuring output signals with an oscilloscope Green trace current output l red trace inverted current output I pink trace TTL output from the Sync Out 74 Analog Output Signals Current output 200 pA and Sync Out Ch2 71 00 V VW To Opal A Cha tov 1 00 V Ch4 5 00 V 10 20 The time lag between the Sync Out output and the current output is approximately 15 us 2 ps if continuous mode is enabled and 50 ps 2 ps if continuous mode is disabled Setup sugg
21. ss zzz Sere ee EE E a amp v cm C Ch Ch3 M ChA M Chs M Che M kom Mm Z7 uim Ready Define the starting point end point and the length of the ramp The ramp is then build according to these parameters in small single steps of 20 us Sine wave pulse type Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 _ Pulse value Unit time Unit value Unit time Unit value Unit time Un afore o mw y m f mv ms pee e epe enn U1 m v 00000360 00 00 00 360 00 00 00 380 00 00 00 400 00 00 00 420 t hh mm zs zzz TE UE ah amp v cni C Ch2 Ch3 ChA Ch5 che T Ready comi Mum E Define the amplitude and the period of the sine wave The sine wave is then computed according to these parameters with a minimum resolution of 1 mV and 20 us If you enter a negative amplitude the sine wave starts with its negative alternation 21 Stimulus Generator 4000 Series Manual 5 5 Autorepeating Pulses and Protocols J pase paue D omt ane nt oe ne tote ene n mnn ocn A4 rectangular n ogg 0 100 0 200 0 300 0 400 0 500 0 ae 0 700 0 600 0 900 1 000 1 100 1 200 E TE Cl e bo gt Chl Che Cho Ch4 Chs f Che Che Che Sweet Senece Sonca For entering complex stimuli easier and faster you can repeat rows and groups instead of entering the pulses several times into the worksheet You can r
22. the length of a current pulse and a high load resistance Current pulse with programmed 16 pA for 100 us and 20 us interval and 1 MQ load resistance The load resistance is very high Please keep in mind that the load resistance of 1 MQ is parallel to 10 MQ of the probe of the oscilloscope That means the load resistance is reduced to 900 kQ only The pulse is biphasic and 16 pA which is a hundredth of the limit of the maximum capacity of the STG4000 series Green trace current output 1 pink trace TTL trigger output from the Sync Out 5 00 V CRhA 5 00V 10 20 Result of Example 3 The current output signal is delivered strongly modified because of the high load resistance of 1 MO and the duration of the pulse 100 us and the interval Before the pulse is reaching its maximum it has to turn into its invert phase Before the pulse is reaching its minimum it is cut off Please see example 4 If the duration of pulse and interval is much longer 1 ms the pulse shape of the delivered signal is much better 89 Stimulus Generator 4000 Series Manual Example 4 Influence of the length of a current pulse and a high load resistance Current pulse with programmed 50 pA for 1 ms and 1 ms interval and 1 MQ load resistance The load resistance is very high the duration of the pulse and the interval are long Please keep in mind that the load resistance of 1 MQ is parallel to 10 MQ of the probe of the oscilloscope That means the load res
23. time of day scale in the format hh mm ss zzz where h means hour m is minute s is second and z is millisecond Under Settings click X Axis Time Format Select the option to enable the time of day scale Deselect the option to switch to the decimal time scale Display Tools Several tools are provided to optimize the graphical presentation of the stimuli If you later choose to display channels the axes of the new displays are set to the default values The time axis can display only positive values The y axis of the Sync Out channels cannot be adjusted because only values of 0 and 1 are available for these channels 45 3 Stimulus Generator 4000 Series Manual Fal ele te a of Enable Scroll the axes by clicking and dragging with your mouse scrolling Enable Zoom the axes by clicking and dragging with your mouse scaling Zoom out Reduces the time scale Zoom in Magnifies the time scale Reset Resets the axes of all channels to the default values Size to fit Adjusts the ranges to fit the largest displayed stimulus 2 re o o Show A horizontal bar is shown that displays the current voltage and the time P4 uiid measure value of the intersection point Drag the bar with the mouse tool Scrolling the axes You can scroll the stimuli forward and backward along the time axis and up and down along the voltage current axis Click es to enable the scrolling Click the axis you like to scroll The m
24. time length of 20 ps Please note that if you work near these limits that is if you use very small amplitudes or very short pulses the output waveform may not exactly match the programmed waveform Resolution Limits Stimulus Output 1 mV 200 nA Ground All stimulation channels of the STG are optically isolated from each other Each channel has an independent ground GND which is usually connected to the ground of the experimental setup for example the amplifier so that all devices refer to the same ground Please make sure that you always use the appropriate ground for the channel in use 69 11 2 Stimulus Generator 4000 Series Manual Voltage Mode If you want to operate the STG in voltage mode you have to select Voltage in Output Mode of the main window of MC Stimulus Il You will usually use the and GND outputs The output signal is identical with the programmed signal in the normal ranges of the electronics accuracy See illustration Voltage mode Standard Setup The output signals are inverted multiplied by 1 if you use the outputs If you use the voltage between and the output signal amplitude is doubled The represents the ground replaces GND in this case See illustration Voltage mode Doubled output signal amplitude A bipolar stimulation is recommended for extracellular recordings from microelectrode arrays MEAs if a very high output signal density in a defined area between two neighb
25. valid batch file for example the sample file TetanusBatch stb in the MC Stimulus II program folder 3 xl Look in E BatchE ample 4m File name TetanusB atch stb Files of type Batch data files stb Cancel Open as read only y The Batch Status Display opens It lists all stimulus files in the batch and the duration of stimulation Batch Status Display Batch File Duration Status c Program Files Multi Channel Systemet MC Stimulus B atehE ample sB aseline stm 00 01 30 c Program Files Multt Channel Systemet MC Stimulus B atchE sample T etanus stm 01 23 25 c Program Files Multt Channel Systemet MC Stimulus 6 atehE sample T esthesponse stm 00 41 00 On the Batch menu click Start to start the download of the first file in the batch list and the STG if the option Start STG after Batch Download on the Settings menu is selected The progress of the pulse protocol is displayed in the Status column of the Batch Status Display Click Stop on the Batch menu to stop the batch if necessary Close the Batch Status Display with the Close command Warning The STG starts immediately after the download if the option Start STG after Batch Download on the Settings menu has been selected Make sure to deselect this option if you do not want to start the stimulation right away Autorepeat option If you select the Autorepeat Batch Mode option on the Settings menu the batch is resta
26. whole columns and autocreate signals You can also right click on a column and select these commands View and Window menu to customize the main window Settings menu to define general settings of the software Help menu to open the MC Stimulus II Help and to display the About dialog where you can find information about the hardware software and firmware versions You will need this information when you contact the support 53 9 3 9 4 Stimulus Generator 4000 Series Manual Toolbar For your convenience you will find some of the more commonly used commands as a button on the toolbar Deal ee amp l ell m New File Creates a new MC Stimulus II file stm file Open File Opens a previously saved MC Stimulus II file stm file Save File Saves the stm file to the chosen destination Cut Row Deletes selected row s and sends deleted row s to clipboard Copy Row Sends selected row s to clipboard Paste Row Pastes row s from clipboard and overwrites row s starting at the selected row Print Prints active channel of active worksheet Information Opens the About dialog where you can find information about the hardware software and firmware versions Download Downloads stimulation file to the connected STG only available if STG has active computer connection Download and Downloads and starts stimulation file to the connected STG only Start available if STG has active computer connection Start Sta
27. you do not see the calibration signals because MC Rack automatically applies a 3 kHz low pass filter during data acquisition and a 10 kHz low pass filter during filter activities When using MC Stimulus II Version 3 3 and higher it is possible to avoid the internal offset calibration Enable the command Disable Autocalibration in main menu STG and download the command to the STG 97 13 Appendix Operating temperature Storage temperature Dimensions W x D x H Weight Fuse Supply voltage external power supply Number of analog output channels Output voltage Output voltage resolution Rise time 10 90 voltage Time lag between Sync Out and voltage output Output current Output current resolution Output current slope Rise time 10 90 current 0 100 pA Time lag between Sync Out and current output 0 100 pA Resolution Time resolution Output signals Maximum frequency rectangular waveform 13 1 Technical Specifications STG4004 and STG4008 10 C to 50 C 0 C to 70 C 325 x 310x 85 mm 2 8 kg internal 3 15 A slow 100 240 VAC 47 to 63 Hz 4 STG4004 or 8 STG4008 8 V to 8 V max x 20 mA 1 mV 1 7 us delta U 8 V 15 2 us amplitude gt 200 mV Standard 1 6 mA to 1 6 mA max 120 V compliance voltage between I and I Or 16 mA to 16 mA max 120 V compliance voltage between I and l Or 160 pA to 160 pA max 120 V compliance voltage between l
28. 0 O02 6O0000 27 20 0s 400002 O02 Os Os Os Off 2l 7002700002 200 20000 O02 6O0000 24 220 400002 O02 Os Os Os D repeat repeat After loading the file into the MC_Stimulus II worksheet you can edit and download the file as usual Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel amp Sync 1 Pulse value Unit time Unit value Unit time Unit value Unit time Unit Row rep Group r i recangue pon mv 20 ms a0 wwo ms oo mWi60 ms 2 2 frectanguer 0 mv me mv js mvo us D 8 recengder 200 mv29 ms 39 mv 20 ms mv ms 2 a recenm o mV 4 ms mv o s mv bs J Dor oor ao OO Dor or do 1 06 00 00 00 200 00 00 00 300 t hh mm ss zzz 00 00 00 400 39 Stimulus Generator 4000 Series Manual Format type 4 This format type is available only in the current version of the import filter It is the plainest format with only one column for the voltage current values and one column for the time lengths This format is the standard format if you want to import data for example from Excel This format type is generated when exporting MC_Stimulus II files as ASCII value time Format type 5 This format type is available only in the current version of the import filter It allows you to use ramp and sine waveforms as well as rectangular waveforms You specify the Pulse type in the first column and the waveform in the three following columns
29. 00 150 200 Cha J I I I I I iecore ee TEE S ss Tes zT Change All Auto Download Reset All Stimulus Intensity dialog box of the 4 channel version of MC Stimulus II Channel 1 is set to 100 95 that is the stimulus protocol will be downloaded onto the stimulus generator as it was programmed in the worksheet Channel 2 is set to 50 95 that is all stimulus amplitudes programmed on channel 2 are halved Channel 3 is set to 150 96 that is all stimulus amplitudes programmed on channel 3 are one and a half Stimulus amplitudes on channel 4 are doubled 33 6 1 ASCII Import Export Loading Files The ASCII import filter is used to load stimulus protocols from an ASCII file into the stimulus worksheet of MC Stimulus Il You can use this feature for feeding recorded signals for example exported from MC Rack see the MC Rack manual into the stimulus generator You can also export the data from MC Stimulus modify it with your custom text or spreadsheet editor and reimport it into MC Stimulus II The current version of the import filter is version 3 1 2 Channel 1 Last Download Output Mode Download Channel Signal Length 3 000 ms Voltage MiC EN max Amplitude 59 000 rmv Ce min Amplitude 182 000 mi urren g EN EN Channel 1 Channel z Channel 3 Channel 4 Channel 5 Channel amp Channel 7 Channel 8 Pulse value Unit time Unit value Unit time Unit value Unit ti
30. 00 pA amplitude is reached The relationship is not linear as shown by the following curve Amplitude Dependency of Rise Time Rise Time ps c 200 400 BD B00 1000 1200 1400 1600 1800 Amplitude HA Resistance dependency of rise time The higher the load resistance the higher will be the rise time The rise time 10 90 was measured with increasing load resistances and a constant current amplitude of 50 pA For 50 pA pulses the relationship is linear up to the maximum load resistance please see Current Mode as shown by the following curve Not shown are the results of the same measurement under the same conditions except of the amplitude with a constant current amplitude of 100 pA Up to a load resistance of 450 kO the results are congruent with 50 pA amplitude an Load Resistance Dependency of Rise Time Rise Time ps bJ DJ RR UR el CI Cc Cl co CI c CI um um 100 200 300 400 500 BOO zoa a00 900 1000 Load Resistance k l Stimulus output comparison The following figures are examples for demonstrating how the amplitude and resistance will affect the kinetics of a current pulse These images document also that in contrast to the current mode in voltage mode the rise time is constant and not dependent on the amplitude or load resistance 78 Analog Output Signals The graphs were generated by a standard oscilloscope The devices were set up according to the recommendations under Voltage Mode an
31. 1 C Off C Current iv5ive amp iv ziv8iliv5sivsEiv iva8 Stimulus protocol Last download time Mode of the STG Channel selection for next download Comment field parameter on the of stimulus protocol active channel page and STG ID Information on the stimulus protocol stim file Autocalibration During the first ten minutes after starting the STG the device emits every minute high frequency signals small bursts for internal offset calibration of the current outputs in a range of 10 mV After warming up the STG the calibration signals later will be delivered half hourly Note It is recommended to start the STG ten minutes before use to allow warming up High sensitive measuring equipments will monitor these calibration signals as artefacts which might disturb the stimulation when using very low stimulus pulses lt 100 pA in current mode Enable the option Disable Autocalibration in STG main menu to avoid the autocalibration Download the command to the STG 4j File Edit ste Batch Signal View Window Settings Help my Start CEHHER osje Dejk Stap CERTET sra Chanel Download Cep jownload Signal Length Downa and Start CErIH max Amplitude wv Disable Autocalibration min Amplitude Select Device Channel 1 pel 4 Channel Pulse e Unit time 13 4 2 Stimulus Generator 4000 Series Manual Operating Multiple STGs lus IT MC 5tim1
32. 2 6 3 7 1 7 2 7 3 Table of Contents Introduction About this Manual Terms of Use for MC_Stimulus II Limitation of Liability Important Information and Instructions Operator s Obligations Guarantee and Liability Important Safety Advice First Use of the Stimulus Generator Welcome to the STG and MC_Stimulus Setting Up and Connecting the STG Driver Installation Installing the Software Operating the STG Operation Overview Operating Multiple STGs 4 2 1 Operating the USB MEA32 STIM4 System Programming Stimulus Protocols MC Stimulus Il Sample Files for STG 4000 Series Neuroelectrophysiology MEA System 5 2 1 General Demo Files MC Stimulus Il Worksheet Pulse Types Autorepeating Pulses and Protocols Autocreating Entries and Editing Columns Auto Sync for Autocreating Trigger Pulses Adjusting the Stimulus Intensity Level ASCII Import Export Loading Files Exporting Files Supported File Formats Stimulus Display Selecting Channels Display Settings Display Tools N N a RWW Uu O O OU Ul 11 14 15 17 17 17 18 19 20 22 25 31 32 35 35 36 36 45 45 45 45 Stimulus Generator 4000 Series Manual 8 2 9 1 9 2 9 3 9 4 9 5 9 6 10 10 1 10 2 10 3 10 4 10 5 10 6 11 11 1 11 2 11 3 11 4 11 5 11 6 11 7 12 12 1 12 2 12 3 12 4 12 5 12 6 12 7 12 8 13 13 1 13 2 13 3 14 Downloading Stimuli Downloading Stimulus Files 8 1 1 Download Modes 8 1 2
33. Compiler Warnings Download Modes Default Mode When downloading files created in MC Stimulus Il software to a stimulus generator of the STG4000 series two different download modes are available In Default mode for each download all channels are stopped and cleared before the new paradigm is downloaded All available channels are downloaded one after the other regardless of the space each channel needs which means that the available memory is optimally used This mode should always be used when large and complicated stimulation patterns are used for example white noise stimulation 49 8 1 2 Stimulus Generator 4000 Series Manual Independent Channel Download The second option is to use the feature Independent Channel Download which is available in the Settings menu This feature allows downloading a new stimulation paradigm to individual channels only while all other channels continue with their former program without stopping To allow this the memory space of the STG is divided among all available channels For example when using a STG4008 the memory is divided in 16 parts for eight Sync Out channels and eight stimulation channels so the memory space for each channel is more limited than in default mode If a stimulation pattern is too large for the available memory partition no download is possible Note It is recommended to operate the stimulus generator in default mode The Independent Channel Download fe
34. GND ground 100 Appendix 13 3 Contact Information Local retailer Please see the list of official MCS distributors on the MCS web site User forum The Multi Channel Systems User Forum provides the opportunity for you to exchange your experience or thoughts with other users worldwide Mailing list If you have subscribed to the mailing list you will be automatically informed about new software releases upcoming events and other news on the product line You can subscribe to the list on the contact form of the MCS web site www multichannelsystems com 101 14 Index 103 Stimulus Generator 4000 Series Manual 104 A About dialog 53 Adjusting Stimulus Intensity Level 32 Amplifier DIGDKITIO d a zi m di eee 57 COHDOCHnQ oia 70 74 Amplitude adjusting stimulus intensity level 32 effects on rise time 77 Of sine WaVG oio seda 20 Analog CUFTent OUtDU E ir eade rl roa 74 MOT uester adds din 8 voltage output esses 70 Arrange Icons 53 ASCII Import without Header 36 importing ASCII files 35 supported file format 36 Assign channels to Auto Sync 31 files to trigger esses 63 trigger inputs to multiple files 63 trigger to channels 60 trigger to Sync Outs
35. Gro up UnqgroUup iiia reu co soupe ra Run Header OT JASCIUTIQ a iet ro Ehre Help I I outputs 7 810101 PEO EETA EOE O ETT 69 74 measuring with an oscilloscope 77 Icons arrange sois tUi ies 53 Ignore files on trigger spena 63 trigger events if channels still active60 Importing JASCII TIGS s executio eco RI d 35 supported ASCII file format 36 Incrementing values in a column 25 Input Ie a 8 MC CaF medtysten ui ocius i 8 MEATOBO er E at a 8 trigger Multi File mode 63 trigger settings oot odes 60 TTE arora ee enter enters er uU se Me 58 Installing SO TWal iis ci edid assed als 10 STG OVET iie E E ds 9 Inverting DUDES oA 25 SOlOCH ON D 25 K Keep Zero 25 L Length OF DUO i ttu tit eet HEEL 20 Load resistance effects ON rise time 77 Loading ASCITES isi oe eet eee 35 files in Batch mode 67 M MC Card CODI ec TIO o eee ose D REDU Ageia 8 MEA1060 CONNECTING gem EE 8 Measure tool 45 Message boxes warning messages eeeeeenn 56 Monophasic pulse 91 Multi File mode 63 Multiple STGs cont olin sa ene b tege ciun er hus 11 Opera UNG arren 14 N New File 54 O Open PAS GU TOS iis ied E E shat 35 gie 54 files in Multi File mode 63 ShOFECUt Key ioie pRdedi aes 54 SUMAS THes coco seven Du EE be 55 Oscillos
36. LEE ERE RE REUE 45 STO a serenaded eececegeneetee 53 Reset All stimulus intensity level 32 Resolution CUFTeID D ost ice oo deuotione iod eder 20 I pp 20 VOITO OC codeine tees 20 Restart channels on a trigger 60 files on trigger seeeeeeeee 63 Rise Time 77 Rising edge OF PEE SION par aede 58 time offset Auto SYNC 31 Row Repeat 22 Run SCHNITT AAE EATA ER ONUS 11 S Save MEC Pere RTT 54 SNOFLCUE KV dia dp uo ient 54 stimulus 1Mes mtt 55 trigger settings seen 60 Scrolling 45 Select Device 11 da igocte 11 0018 4 0 UPPER NE CPU MP TEC NEOEM 11 Serial port 8 Settings ZAUtO S WC hia ich m Ed 31 MENU PRSE T 53 Multi File mode 63 SVDEIUT Loa dv E uM e t 60 dg ole RH errr er orre erreer en Toe r eee 60 shielding CADIOS anaa aes 70 74 Shifting Vall OS eeoa braun de dub eed bane 25 Show Auto Sync output 31 measure LOO io Re uns 45 Signal o Yoidiors uc 57 Current mode ei a D terius 74 measuring output signals with an OSCIITOSCODG xi swiss vrbes ec Erie desx cube ae 77 MENU seru gu De cu ut a 53 SVDICAU sorta ane debui eu o i 57 voltage mode sss 70 Stimulus Generator 4000 Series Manual Sine wave pulse type eieren 20 Single Trigger 60 Size to fit 45 Software installa
37. Liability Multi Channel Systems MCS GmbH makes no guarantee as to the accuracy of any and all tests and data generated by the use the MC Stimulus II software It is up to the user to use good laboratory practice to establish the validity of his findings To the maximum extent permitted by applicable law in no event shall Multi Channel Systems MCS GmbH or its suppliers be liable for any special incidental indirect or consequential damages whatsever including without limitation injuries damages for data loss loss of business profits business interruption loss of business information or any other pecuniary loss arising out of the use of or inability to use MC Stimulus Il or the provision of or failure to provide Support Services even if Multi Channel Systems MCS GmbH has been advised of the possibility of such damages 2 1 2 2 Important Information and Instructions Operator s Obligations The operator is obliged to allow only persons to work on the device who are familiar with the safety at work and accident prevention regulations and have been instructed how to use the device are professionally qualified or have specialist knowledge and training and have received instruction in the use of the device have read and understood the chapter on safety and the warning instructions in this manual and confirmed this with their signature It must be monitored at regular intervals that the operating personnel are working safely
38. Measuring output signals with an oscilloscope Green trace voltage output U red trace inverted voltage output U pink trace TTL output from the Sync Out Voltage output 1 V and Sync Out Ch2 Soomv SH T0 OMS A CHIP TE s 0mVv Chg 5 00 V 10 20 The time lag between the Sync Out output the trigger and the voltage output is approximately 15 ps 2 ps Setup suggestions for voltage mode The following illustrations show circuit diagrams of suggested setups Please note that you can use unshielded twisted pair cables wires twisted round each other instead of a shielding to prevent the amplifier from picking up noise The experimental ground can be for example the amplifier s ground or a Faraday cage If you use the stimulus generator together with a standard MEA amplifier you can also use the provided 3 polar cable for directly connecting the stimulus generator to the electrode inputs of the MEA amplifier Please note that a shielded cable is generally not necessary if you use a MEA1060 amplifier with blanking circuit For more details on the recommended setup for this amplifier type please refer to the MEA1060 Amplifier with Blanking Circuit User Manual 72 Analog Output Signals Yoltage mode Standard setup Stimulus generator outputs MEA amplifier inputs Shielding Ground Electrode Voltage mode Doubled output signal amplitude Stimulus generator outputs MEA amplifier inputs Shiel
39. SB connector to the USB port of the computer The computer connection is necessary for programming the STG but not for operating it except in Batch mode Connect the and plugs of the required output channels with 2 mm plug cables to the stimulating electrodes Connect the appropriate ground to the ground input of the stimulation setup for example to the ground input of the MEA1060 amplifier Current and voltage driven stimulation is generated from the same outputs Optional Connect the required Trigger In inputs with BNC cables to instruments that produce TTL signals for triggering the STG Optional Connect the required Sync Out outputs with BNC cables to following instruments that you want to be triggered by TTL signals from the STG for example to one of the 16 digital inputs of the MC Card Switch the STG on by pressing the toggle switch on the rear panel Note It is recommended to start the STG ten minutes before use to allow warming up 3 3 First Use of the Stimulus Generator Illustration of the STG 4000 series front and rear panel with connectors STG 4004 Front Panel STG 4004 multichannel 1 gt systems TUO T z0 POWER 75 GND amp GND PROG C SWEEP 5 START STOP STG 4004 Rear Panel cineca SYNC OUT TRIGGER IN 0o 92000 0000 STG 4008 Front Panel STG 4008 multichannel O 1 3 8 systems 0 0 POWER O GNDO GNDO GND PROG 7 0 0 0 SWEEP START STOP
40. TG will light up but the brightness of the LED is not as brilliant as in running state 61 Stimulus Generator 4000 Series Manual Select Single Trigger if you want to start the stimulus protocol once via external trigger trigger event while sweeping on channel assigned to same trigger Is Stop C Restart C Ignore Trigger as Gate Single Trigger Set up your stimuli with the MC Stimulus Il program Download the stimulus protocol from a MC Stimulus II file to the STG Arm the STG either manually by pressing the Start Stop button on the front panel or with software controls To arm the STG via software use the start icon Sweep S a in MC Stimulus Il main menu or click the combined LED Start Stop button EN EE ENS NN for the Sweeps When armed the STG accepts one single trigger on its trigger input The trigger input must be connected to the device which generates the trigger To start the stimulus protocol a second time the STG needs to be armed again manually As long as the STG is armed the LED in MC Stimulus Il appears blue to indicate the armed state The stimulation will automatically be started after applying the single trigger from the external device The LED in will turn to red while the trigger is running and return to grey after the trigger has finished the stimulus protocol The STG needs to be armed again to accept the next trigger F MC Stimulus II MC Stimi Fie Edit STG B
41. The general rules for setting up ramp and sine waveforms fully apply see also Pulse Types The pulse types are defined by the following numbers Rectangular 0 ramp 1 sine 2 You can only use the rectangular type 0 for the Sync Out channels Important When setting up rectangular or sine waveforms in this format type you need only one value column for defining the amplitude but you have to define both value columns even so The first value column is ignored for rectangular and sine pulse types by the MC_Stimulus II program pulse 0 1 2 value value time 40 Example This is an ASCII file viewed in a standard text editor program You can see the tabs spaces and CR LF ASCII Import Export For demonstration purposes a ramp type pulse was programmed on the first channel a sine wave on the second channel a rectangular waveform on the third channel and mixed pulse types on the fourth channel L file exanple datY z Copyright ic 2005 Multi Channel Systems H this is an example file for MC Stimulus ASCII import3T 3 4 5 e 7 S 9 10 JE lz l3 l4 s 16 l7 l3 13 20 el ez 23 ed 2s 26 e es z9 30 3l ci 33 34d 36 36 37 38 39 40 4l dz 43 Hulti Channel Systems HMc stimulus ASCII import ersion 1 1097 this is a coment lined output mode voltaged aq 1d value enu 4000 2000s 1000 eT value SOOO 1 0000s UO 33 value
42. _Rack The STG generates voltage pulses in values of millivolt Please use the Amplification factor of the ASCII Import Dialog to convert uV in mV otherwise the output of the STG is zero Amplification 10000 Cancel The amplification factor is generally applied in files with and without header 43 Stimulus Generator 4000 Series Manual ASCII Import of Files For ASCII import of files please use the command ASCII Import in main menu Settings F MC Stimulus IT MC_ Stimulus Main Menu File stm File Edit STG Batch Signal View Window Settings Help Enable compiler warnings L Ga Ez Y Dl cb E T w Start STS after Batch Download F MC_Stimulus_Main_Menu_File stm Autorepeat Batch Mode L w A Axis Time Format abis h sin ar Auto Sync gnal Length ms iN mas Amplitude 0 000 rv r evice Serup sse 10 GIRO pa SEN reli The dialog ASCII Import Settings will open ASCII Import Settings X Settings For ASCII import Files without header lines Format x From row 3 v Output Mode Voltage C Current Amplification on Settings for ASCII import files without header Select the format type from one to six from the drop down menu Format Select the number of the row from which the file should be imported Row numbers are available from one to ten from the drop down menu From row Select the Output Mode in the check boxes Voltage or Current Settings f
43. ader to select a column The selected column is highlighted in black 2 Right click and click Change Time Unit annel 6 Channel 7 Channel 8 Sync Increment selection Guborill Group Ungroup Set Time Unit Cancel All units in the column are set to the selected time unit annel 6 Cha Inverting pulses in a column 1 Click any Value column header to select a column The selected column is highlighted in black 25 Stimulus Generator 4000 Series Manual Right click and click Invert Selection Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 Sy Puise value unt time Unit value Tunit time unit value unt time Unit Row repeat Group repeat T jredenue poo mv 0 me MENS Concent mien ms SSS 3 rectangular 0 Increment selection Ve mw e a sne 20 Autofil mwe ims 5 rectang o me Bjrecenue eren Unto mus z recangder mw e im Dr ad ad dao Do ad ad 2060 Do an ad 4060 Dr ad ad eo 00 00 00 200 00 00 07 000 Bo adr 1 200 t hh mm zs zzz The polarity of the signals is switched from positive to negative and vice versa Carmeli 5yr J Puse value unit time unit aiue una time Unit vatue Uni time Unit Row repeat Group repeat 3 rectangular 0 NM o0 ow 05 a fsine 20 S frectanguar
44. age mode sss 70 Undo SDOFECUT KEV iioaseeasedpus eoa donatae 54 Update SOTLWOFG ceca dnte undue to dede p aee 10 USB connector 8 USB MEA32 STIM4 System 14 Select Stimulation Channels 14 V View MEN U nerina 53 Voltage maximum output 70 99 measuring output with an oscilloscope 77 output mode 19 69 70 l OSOIU TIO isses cus ood oet etu odi dads 20 W Warning messages CISAD WING erran a ie 49 Window VIG ssooxedeos stet a de dastivu deant 53 WYSIWYG window 45 Z Zooming 45 109 110
45. all channels In current mode Connect the unused current output to ground GND Please see also the recommended setups in the chapter Current Mode Output Signal Does not Match Programmed Signal The amplitude or the duration of the output pulses differs from the programmed signal Possible causes The stimulus protocol works near the resolution limits of the STG that is the pulses are very short or the amplitudes very low smaller than 50 mV or 10 pA Check the output signals with an oscilloscope and modify the pulse protocol so that the amplitude and timing of the pulses will match your requirements You operate the STG in current mode and the electrode resistance of the stimulating electrodes is too high According to Ohm s law the electrode resistance directly affects the kinetics of the current output The rise time increases with an increasing electrode resistance If the electrode resistance is too high for the compliance voltage the output signal will be clipped Try to stimulate in voltage mode Try to use stimulating electrodes with a lower impedance Check the output signals with an oscilloscope and modify the pulse protocol so that the amplitude and timing of the pulses will match your requirements 96 12 7 12 8 Troubleshooting Signal is not Repeated Pulse Train Fails The STG fails to generate a train of pulses When a pulse is programmed in MC_Stimulus and then repeated in the Trigger Settings dia
46. and l Standard 200 nA with 1 6 mA Or 2000 nA with 16 mA Or 20 nA with 160 A 600 pA us R 10 ko 1 4 us R 10 ko 15 2 us QR 10 kQ if continuous mode is enabled 50 2 us R 10 kQ if continuous mode is disabled 14 bit 20 us Freely programmable rectangular ramp sinusoidal 25 kHz Memory capacity for stored 64 MB waveforms Timing on external trigger Minimum time length of trigger input TTL 40 us Time lag between trigger input and Sync Out 70 10 us Maximum trigger input frequency 10 kHz 99 13 2 Stimulus Generator 4000 Series Manual Interface and connectors Sync Out for synchronization with following devices 4 8 digital channels via coax cable Trigger In for synchronization with preceding devices 4 8 digital channels via coax cable Interface connection to computer USB 2 0 High Speed Download rate max 480 Mb per second Software MC Stimulus II Version 3 3 2 and higher Operating system Microsoft Windows 7 and 8 Vista or XP with NTFS English and German versions supported Data import export ASCII file format Note Other configurations of output currents are available on request Please contact MCS Pin Layout Digital I O Pin layout of the DIGITAL IN OUT connector Available with STG4004 and STG4008 25 Pin Female D Sub Connector Pin 1 8 Digital input DO D7 for Extended Multi File Mode Pin 9 16 Digital output D8 D15 not connected Pin 17 25
47. and the open end of the cable from the GND output with the ground cable from the probe This means you measure the potential difference between the U output and ground Please note that you need a resistor in parallel to measure the current output properly because oscilloscopes measure voltages If the input resistance of the oscilloscope is too low the voltage output of the STG will be small as well and you will not see any signals on the oscilloscope MCS recommends a resistor of 10 kQ because the output signal of the current output should then be identical to the output signal of the voltage output You can use the same test protocol regardless of the output mode for the current and voltage outputs and compare the signals As an alternative you can also use a resistor that mimics the electrode resistance for testing your experiments Choose current mode in the main menu of the MC Stimulus Il software What you do is you plug in cables into the I output and into the GND output of the STG You then use alligator crimps for connecting a 10 kQ resistor between the two open ends of the cables You then touch the cable on one side of the resistor with the probe and on the other side with the ground of the probe Alternatively use the provided cable with integrated 10 kQ resistor That is you measure the potential difference over the resistor You can then calculate the current output according to Ohm s law U R I For example if you stimul
48. arres MEAE EEEE 00 xffamnets smer smee syncs Puise watue unif time unit DK IH rectangular 100 E Increment selected values eis 2 rectangular 100 mw E bu qui x Cancel I3 rectangular 100 my HE a rectangular 100 my Us Tupe of Increment I5 rectangular 100 m amp I5 rectangular 100 mv H Cf Count 5 a Keep er C Shift C Absolute value Dor ad ad dao Dr an adr 01 t hh mm ss zzz The value in the first row is incremented by the selected start value of 10 The next value is modified by the double value 20 and so on Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel amp Channel 7 Channel 8 mum 0 umo Sc mmo SS mda Lemos o mS mw 0 5 mw o5 j zi mv s 30 4 rectangular 100 mv EW S jrectenguer 100 mv Sg niu cian nn ZH 2 L ZH 28088 MI An Dor ad ad dao Dr an ad a1 t hh mm ss zzz 28 Programming Stimulus Protocols You can now fill further columns in the same way Thus you can easily set up a stimulus with incremented pulse lengths like this with only a few mouse clicks Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 Pulse value Unit time Unit value Unit time Unit value Unit time Unit Row rep Group r d frectenguer 0 mvjo gs aoo m so e mv w mv 20 js jroo m s e m s 3 recangier i0 mv 3 400 mw e0 e mW
49. atch Signal View Window Settings Help ojaa ee e vis e m STG 20020 Disconnect m Programmed Sweep E S It is possible to stop the stimulation at any time while the STG is armed or is running via Start Stop button on the front panel of the STG or via software Sweep Count The number of sweeps are counted and displayed for each of the triggers at the bottom of the main window V Synet Sweep Count Trigger Number Ta 4062 0060 Mmi Please note that not the number of stimuli but the number of sweeps is counted whereby one sweep is defined as one complete run of the programmed stimulation pattern as displayed in the main window So if five stimuli are programmed and the repeat rate in the Trigger Setting menu Is set to six 30 stimuli will be delivered but the sweep count will be six Number of sweeps 1 100 2 110 3 120 4 130 Sweeps are not counted online the final number is displayed after the STG has been stopped The number of sweeps per channel is not put to zero automatically every time the stimulation is started Instead a reset only happens if a trigger is again used in the new stimulation So if a stimulation file uses trigger one and two with a repeat of three each the display will show 1 3 2 3 3 0 4 0 at the end of the stimulation If afterwards for example a stimulation file is started which uses only trigger four in continuous mode and this file is stopped after 15 repeats
50. ate for your application and stimulating electrode type For using MEA electrodes as stimulating electrodes MCS recommends the use of monopolar voltage pulses though some MEA users feel that they have better results with bipolar voltage pulses depending on the cell culture or tissue used Warning Always obey the recommendations of the stimulating electrode manufacturer Neuroelectrophysiology MEA System LTP Induction stm Stimulus type Pulse train of 100 biphasic pulses for LTP induction Amplitude 1 5 V Duration 100 us per phase Inter Stimulus Interval Frequency 10 ms 100 Hz LTP Testpulse stm Stimulus type Biphasic test pulse Amplitude 1 5 V Duration 100 us per phase Inter Stimulus Interval 60 s PairedPulse stm Stimulus type Paired biphasic test pulse for paired pulse facilitation and depression Amplitude 2 V Duration 100 us per phase Inter Stimulus Interval 20 ms between paired pulses 10 s between pairs lOcurve 500 3000 stm Stimulus type Voltage step series with biphasic pulses for I O analysis Amplitude 500 mV to 3 V increments of 250 mV 3 repeats Duration 100 us per phase Inter Stimulus Interval 10 s 17 5 2 1 Stimulus Generator 4000 Series Manual monophasic_voltage_1V_100ps stm Stimulus type Monophasic test pulse Amplitude 1 V Duration 100 us Inter Stimulus Interval 24 ms biphasic voltage 1V 100ys stm Stimulus type Biphasic test pulse Amplitude 1 V Duration 100 u
51. ate with 100 pA the voltage over the 10 kQ resistor has to be U 10000 Q 0 1 mA 1000 mV 1 V The following illustration shows four possible combinations of connecting the current outputs on the front panel of the stimulus generator to the oscilloscope Again it is important that unused current outputs are connected to ground Measuring current output signals with an oscilloscope to GND to GND Difference Rise Time The rise time of the voltage output is constant and independent from the stimulus protocol The rise time of the current output is no fixed hardware property The kinetics depend on the amplitude of the current pulse and the load resistance that is generally the electrode impedance The resistance affects the kinetics much stronger than the amplitude Generally this does not make much difference for biological applications If you have very time critical applications or if you use very short pulses you should check the output of the STG with an oscilloscope before starting the experiment 77 Stimulus Generator 4000 Series Manual Amplitude dependency of rise time The rise time is independent from the current amplitude in the common range of amplitudes The rise time 10 96 90 96 was measured with increasing current amplitudes 100 pA to 1600 pA and a constant load resistance of 10 kQ The output current is nearly constant up to amplitudes of 1100 pA but increases about 0 3 us until the maximum of 16
52. ature should only be used if one STG is connected to more than one recording system and if those recording systems should be operated independently Independent Channel Download The Independent Channel Download can be activated in the Settings menu As mentioned above it allows downloading of new paradigms to individual channels without stopping all other channels but also leads to limited memory availability F MC Stimulus II MC_5timi1 4 Fie Edit STG Batch Signal View Window Settings Help C zj X Ea e T w Enable compiler warnings W Start STG after Batch Download Channel LastD y Autorepeat Batch Mode Signal Length U ys w 5 Axis Time Format max Amplitude p w Auto Syne min Amplitude p ST Device Setup Ch 1 Ch l2 Ch 3 Ch Sea anne anne anne ann w Independent Channel Download This feature is primarily intended to be used with MEA Systems with multiple amplifiers and only a single STG See chapter Operating multiple MEA amplifiers with one STG If the independent download option is active a click on the Download or Download and Start button will only download a new paradigm to the channels selected in the Download Channel check box 1 and 6 in this example Other channels will not be influenced Download Channel Mirar 5 amp 8 However the individual download option is influenced by the Trigger Settings To be able to download channels independently from
53. box whether you changed them all or not Download Syne wil 2f3f 4 gH sg EN E Download Channel 1mwm2apspa IE EN EN E Only these channels will be downloaded ongoing stimulation on other channels will not be interrupted To start and stop only the channels associated with one amplifier use the Start Stop buttons for individual triggers Each button controls one trigger and hence all assigned channels The orange LED indicates ongoing stimulation In our example the configuration shown below would mean that amplifiers A B and D are receiving a stimulation signal while stimulation for amplifier C is stopped Triggers 5 to 8 have no function in this example You could download a new paradigm for amplifier D by downloading channels 5 and 6 and Sync Out 3 and then start it with the start button of Trigger 3 This would not interfere with the ongoing stimulation to the other three amplifiers but only if the Independent Channel Download function is activated If the STG is in default download mode any download will stop all ongoing stimulation and empty the memory of the STG See chapter Downloading Stimulus Files for more information 52 9 1 9 2 General Software Features General Software Features Customizing the Main Window You can customize the size and position of open file windows and the toolbar in MC Stimulus II You can hide the toolbar and the status bar by deselecting them on the View menu You ca
54. capacity of the STG 4000 series Green trace current output l pink trace TTL trigger output from the Sync Out 10 0V amp Ehd 5 00V i 10 20 Result of Example 1 The current output signal is delivered unmodified without any influence of the length of the pulse with an amplitude of 1600 pA which is at the maximum capacity of the STG 4000 series 86 Analog Output Signals Example 2 Influence of the length of a current pulse 100 ys versus 20 us with an amplitude of 16 uA which is a hundredth of the limit of the maximum capacity of the STG 4000 series constant amplitude and load resistance Example 2a Current pulse with programmed 16 pA for 100 us and 20 us interval and 10 kQ load resistance The pulse is biphasic 16 pA Green trace current output l pink trace TTL trigger output from the Sync Out 200mV amp E hd 5 00 V 10 20 87 Stimulus Generator 4000 Series Manual Example 2b Current pulse with programmed 16 pA for 20 us and 20 us interval and 10 kQ load resistance The pulse is biphasic 16 pA Green trace current output l pink trace TTL trigger output from the Sync Out 200mv Ch4 5 00V i 10 20 Result of Example 2 The current output signal is delivered unmodified without any influence of the length of the pulse and an amplitude of 16 pA which is a hundredth of the limit of the maximum capacity of the STG 4000 series Analog Output Signals Example 3 Influence of
55. ck on the Help menu click About The software SW and hardware HW version you are currently using On the Help menu click About The displayed dialog box shows the version numbers The operating system and service pack number on the connected computer The hardware configuration microprocessor frequency main memory hard disk of the connected computer This information is especially important if you have modified the computer or installed new hard or software recently 93 Stimulus Generator 4000 Series Manual 12 3 Error Messages This chapter explains error messages to you that may occur during normal operation They do not present a reason to worry and can be easily avoided General operation Can nat close C Program Files Multi Channel SystemsiMc Stimulus ITpemoao 1 stm It is a Multi File Mode File is displayed when you try to close a file in Multi File mode You cannot close files if you are in Multi File mode indicated on the MC Stimulus status bar Please leave the Multi File mode by selecting Standard under Mode in the Trigger Settings dialog box before closing files Download These error messages may occur during download on the STG Please note that you can switch off some of these messages if they disturb your experiment See chapter Downloading Stimuli for more details on downloading MCStimulus NT The worksheet is empty No data is present that zd can be downloaded Please create a stimul
56. cope comparing STG current signals 85 measuring current signals 74 measuring STG output with an OSCIIIOSCODG co sosta oe ERI OR adu EU 77 measuring STG signals 70 Output analog output signals 69 autocreating Sync Out pulses 31 doubling amplitude 70 measuring with an oscilloscope 77 DIOU usi at o de eno 19 signal inversion eee 70 74 Sync Out settings ssssss 60 P Panel year panel csdotnete am pen epo ecc iotie us 8 Paste qs sees 54 ShOFPICUE KOy iiber p List ind 54 Period of sine Wave sisarien nenne 20 Port QUOT al tt 8 SOM ae RNC aa 8 USB MTM een RO UNDE TNNT 8 Post Time 31 Pre Time 31 Print Previo W C E 55 printing worksheet 55 SOUD MT ER 23 SNOMLCUL KEV oaia i se sus d 54 Pulse Autocreating Sync Out pulses 31 CUI OtIOD iss o MEE zoe DE RE PERMEERR 20 leng t E tsscecesetesdtoliioen metam unes tacts indes 20 EVD CS itch eis Rosie LA d Lr pude ERU 20 R Ramp pulse tVDE uocat seii CoA I ES 20 Rear panel 8 Rectangular pulse TDG inar EFE E EERRRE IV 20 Repeating a group Of l OWs ssseseeeeennnnn nn 22 GONTINUOUSIY cus pudo a Ua doe etu uH 22 OV TIO OCF gem p 22 ISOS e ht eta nen rene aes mens 22 Qe 22 Requirements SYSTE ox cesteduisto ais a de ERI RUM RN 10 Reset GISDIGV isis use RP EMB
57. d Current Mode The same pulse protocol and the same channel number of the STG outputs were used for measuring the voltage and the current output Example 1a Voltage Mode The green trace shows the voltage U output and the pink trace shows the TTL output from the Sync Out that was used for triggering the oscilloscope Rise time with programmed 500 mV and 50 pA pulses 10 kQ load resistance Rise time 10 90 909 2 3 us H4 00us A Chd J 1 60 V 200mV hd 5 00 V ij 10 20 79 Stimulus Generator 4000 Series Manual Example 1b Current Mode The green trace shows the current output I with a load resistance of 10 kQ and the pink trace shows the TTL output from the Sync Out that was used for triggering the oscilloscope Rise time with programmed 500 mV and 50 pA pulses 10 kQ load resistance Rise time 10 90 909 2 3 us H4 00us A Chd J 1 60 V s00mV Chg 5 00 V ij 10 20 Result of Example 1 The rise time is about 3 us for the 500 mV voltage pulse and about 3 us for the 50 pA current pulse 80 Analog Output Signals Example 2a Voltage Mode The green trace shows the voltage U output and the pink trace shows the TTL output from the Sync Out that was used for triggering the oscilloscope Rise time with programmed 5 V and 500 pA pulses 10 kQ load resistance Rise time 10 99 90 909 2 3 us 2 00 amp EChd 5 00V 10 20 81 Stimulus Generator 4000 Series Manual Example 2b Cu
58. d be immediately followed by an inverse signal of the same area product of current and time which helps to discharge the electrode The easiest way is to use the same signal amplitude with an opposite polarity Multi Channel Systems recommends using voltage driven stimulation In voltage mode it is not necessary to use biphasic stimulation for discharging the electrodes because the electrode should be discharged at a voltage of 0 that is it would be sufficient to apply the negative phase only Warning When using MEA electrodes of TIN material stimulation with higher amplitudes is possible if you apply the negative pulse first Also regard the safe charge injection limit as described in the MEA manual Otherwise electrodes can be damaged during stimulation The following illustration shows the effect of a biphasic current pulse on the discharge of the stimulation electrode As you can see the first monophasic pulse is followed immediately by a pulse of the opposite polarity and the same product of current and time Effect of a bipolar current pulse on the electrode voltage Monopolar pulse Bipolar pulse Simus Ec LA T NZ IL Y dL V 91 12 12 1 12 2 Troubleshooting About Troubleshooting Most problems occur seldom and only under specific circumstances In most cases it is only a minor problem that can be easily avoided or solved If the problem persists please contact your local
59. ding Electrode Ground Voltage mode Dipolar stimulation Stimulus generator outputs MEA amplifier inputs A Electrode 1 Shielding Electrode 7 Ground 73 11 3 Stimulus Generator 4000 Series Manual Current Mode If you want to operate the STG in current mode you have to select Current in Output Mode of the main window of MC Stimulus Il You will usually use the and GND outputs See illustration Standard setup The output signals are inverted multiplied by 1 if you use the outputs If the load resistance that is the electrode impedance is very high the output voltage can get very high too The stimulus generator limits the output voltage to 75 V between or and GND The guaranteed compliance voltage is 60 V that means 120 V between and The current output of the STG is limited by the compliance voltage and the electrode impedance If the maximum load resistance for a given current output is exceeded the compliance voltage 60 V between I and GND of the STG will not be sufficient for delivering enough current and the output will be clipped You can calculate the maximum load resistance by Ohm s law For example if you want to program a 100 pA pulse the maximum load resistance will be 600 kQ According to Ohm s law R2U 1260V 100 pA 600 KO that is 60 V are sufficient for delivering a current of 100 pA at a load resistance of 600 kQ Therefore if you use current driven stimulation you
60. each other it is mandatory that these channels are assigned to different triggers If a new paradigm is downloaded to one channel all channels assigned to the same trigger will also be stopped even though no new paradigm is send to this channel Each time you change any parameter in the Trigger Settings dialog the running stimulation will be stopped automatically You have to start the stimulation again manually 50 General Software Features The following dialog appears when you change anything in the Trigger Settings dialog Click Yes button to download on all channels click No button to download on selected channels MC Stimulus il h Channels which are not selected will be cleared F Do you want to download all channels 8 2 Operating multiple MEA Amplifiers with one STG It is possible to operate more than one MEA amplifier independently with a single stimulus generator of the 4000 series To do this activate the Independent Channel Download feature in the Settings menu F MC Stimulus II MC_Stimi 4 File Edit STG Batch Signal View Window Settings Help D d X ESE p w Enable compiler warnings W Start STG after Batch Download Channel LastD y Autorepeat Batch Mode Signal Length OU ys w 5 Axis Time Format max Amplitude p w Auto Syne min Amplitude p ST Device Setup Ch 1 Ch l2 Ch 3 Ch OE anne anne anne ann w Independent Channel Download This will allow you to dow
61. enu click Export ASCII File The Save As dialog box appears Browse to the target folder and enter a file name Click Save to generate the ASCII file All pulse protocols from the active file will be saved in ASCII file format The MC Stimulus II ASCII format type is format type 4 For your convenience the generated ASCII file includes the header that is required for reimporting the file into MC_Stimulus II Supported File Formats The ASCII text must exactly follow the specifications below The parameters in angle brackets are placeholders for any valid numbers or strings Hint You can set up files conveniently in any standard spreadsheet program and save them as tab delimited ASCII General The file name and the file name extension are not important You can use any extension you like for example txt or dat or any other Comments are introduced by the number sign The rest of the line after is only for your information it is completely ignored when executing the stimulus file You do not need comments of course but it is convenient for documentation Hint Use to comment lines out That is you can disable lines without removing it from the file Thus you can keep the lines for later uses Blank lines are ignored The spelling is strict The MC Stimulus Il program does not recognize a keyword if it is misspelled or if even only one character is missing but it is not case sensitive The file has the following s
62. epeat each single row In the preceding example the rectangular pulse in row 1 is repeated twice You can also group several rows and repeat them altogether In the example pulses from rows 1 to 4 are grouped and repeated three times Repeating rows Type the number of repeats into the Row Repeat cell of the appropriate row The pulses of the complete row are repeated the specified number of times Grouping and repeating groups 1 Select several cells by clicking and dragging cells in the Group Repeat column Selected cells are highlighted in black 2 Point to the selected cells and right click to open the context menu STL IL Group repeat peat Change Time Unit Invert selection Increment selection Generate Franm Ta Bsy Group Ungroup 3 Click Group Ungroup The corresponding rows are grouped now This is indicated by a merging of the Group Repeat cells 22 Programming Stimulus Protocols 4 Type the number of repeats into the merged Group Repeat cell at Group repeat Repeating complete pulse protocols autorepeat function Continuous Mode You can repeat the complete stimulus on all output channels that are triggered by the same trigger This means each time when a trigger event occurs the complete stimuli on all corresponding output channels are repeated for the specified number of times If the stimuli have different lengths the starting points are synchronized For example
63. er Sync1 If you use a MEA1060 BC amplifier you can use up to two output channels per amplifier that is you may want to select Ch1 and Ch2 for Sync1 The preceding screen shot shows an advanced setup that could be used for example for a setup with 4 MEA1060 BC amplifiers each connected with two analog STG channels and one Sync Out Set the time offset of the TTL s rising edge before the stimulus pulse Pre Time and of the TTL s falling edge after the pulse Post Time For operating a MEA1060 BC amplifier a Pre Time of 0 us and a Post Time of 100 ys are default but especially the Post Time may have to be adjusted according to your stimulus amplitude and the stimulus artifact suppression performance please see the user manual of your MEA1060 BC amplifier or the MEA Select Help for more details On the Settings menu click Auto Sync to enable the Auto Sync feature The Sync Out protocols will be autocreated in synchrony with the pulse protocols on the assigned channels will show up in the WYSIWYG window immediately and will be downloaded onto the STG with your next download Please make sure that the display and download options are appropriate that is all necessary Sync Out channels are selected The worksheets of the Sync Out channels assigned to the Auto Sync feature will be unavailable that is you cannot edit them manually The following screen shot shows an example of a monophasic 100 us voltage pulse on channel 1 and the corresp
64. er output sweep than the other and is not active anymore 60 Synchronizing Events Triggered action when channel s still active You can define what action should be taken when any of the channels assigned to the trigger are still active Please click the expand button Y to show the advanced options trigger event while sweeping mon channel assigned to same trigger Stop C Restart Ignore Trigger as Gate C Single Trigger __ Cancel Stop A following trigger input stops the stimulation on the assigned channels if at least one channel that has been assigned to this particular trigger is still active in the moment of the trigger event The trigger event starts stimulation only if no channel that has been assigned to this trigger Is active Restart The stimulus protocols on all channels assigned to this trigger are started at the trigger event regardless of the activity on these channels Ignore The trigger event is ignored when any channel assigned to this trigger is active The trigger event starts stimulation only if no channel that has been assigned to this trigger is active Use Trigger as Gate Channels associated with a Trigger input will be active as long as the condition of that trigger is 1 HIGH and switched off as soon as the condition becomes 0 LOW If the stimulation protocol is shorter than the gate it will stop at the predefined time regardless of the gating trigger If the stimulation prot
65. estions for current mode The following illustrations show circuit diagrams of suggested setups Please note that for current stimulation you should use shielded cables to protect your setup against electromagnetic interference The experimental ground can be for example the amplifier s ground or a Faraday cage 75 Stimulus Generator 4000 Series Manual Current mode Standard setup Stimulus generator outputs MEA amplifier inputs Shielding Electrode Ground Current mode Double maximal compliance voltage Stimulus generator outputs MEA amplifier inputs Shielding Electrode n Current mode Dipolar stimulation Stimulus generator outputs MEA amplifier inputs Electrode 1 Shielding Electrode Ground 76 Analog Output Signals 11 4 Measuring Output Signals with an Oscilloscope 11 5 For test purposes you can measure the output signals with an oscilloscope Please see the appendix for the pin layout of the sockets on the front panel of the STG4000 series Most standard oscilloscopes have a probe with a ground cable This probe is usually connected to the oscilloscope via a BNC socket What you do for measuring the voltage output is the following Choose voltage mode in the main menu of the MC Stimulus II software You plug in the standard STG cables into the U output and the corresponding GND output You then touch the open end of the cable from the U output with the probe from the oscilloscope
66. ful if you want to start a sequence of different stimulation protocols one after the other on the same trigger Trigger 1 Please make sure that you have assigned all channels to Trigger 1 in the Trigger Settings dialog box You set up the protocols and save them as separate stimulus files Then you set up a batch file with a list of the desired stimulus files e The first trigger event starts the first file in the list e When a file has been completed the next file in the list is downloaded automatically e When a trigger event occurs after the file has been run the next file is started and so on e When a trigger event occurs during the run of a file the run is stopped and the next file in the list is downloaded The next trigger event starts the next file e Deselect the option Start STG after Batch Download on the Settings menu if you want to start the protocols on a trigger or manually with the software or hardware controls If the option Start STG after Batch Download on the Settings menu is selected the STG starts each file directly after the download without waiting for a trigger event As an alternative to the Batch mode you can also use separate trigger inputs to trigger separate output channels or separate stimulus files This way you can trigger up to four different pulse protocols on separate channels See chapter Triggering Stimulation for more details Note Only Trigger 1 is available for triggering in Batch mode Do
67. g box the corresponding Sync Out signal is repeated as well as the selected analog output signals You can for example synchronize four separate displays or other virtual instruments in MC Rack one for each trigger input You can also define a bit pattern in the Trigger Detector and trigger instruments on this bit pattern for example to activate virtual instruments only when trigger 1 and 3 are active at the same time Please see the MC Rack help or manual for more information on the Trigger Detector and triggering Note No Sync Out signal is sent when the STG is stopped on a trigger event or by pressing the Start Stop button that is if the trigger event occurs or the button is pressed when the STG has still been active Timing of blanking signal If you have a MEA amplifier with blanking circuit in use you can apply a blanking signal TTL pulse for transiently switching off the input stage of the amplifiers during the stimulus thus avoiding the stimulus artifact MCS recommends a minimum blanking pulse that starts with the stimulus and stops 100 ps after the stimulus The fact that the digital output of the stimulus generator from MCS is about 15 ps faster than the analog output ensures that the blanking signal is delivered before the stimulus It is very important that the blanking pulse is delivered shortly before the stimulus 57 10 2 Stimulus Generator 4000 Series Manual The following example shows a blanking signal pr
68. ger input 1 30 10 us Custom switch for remote controlling of the STG You can connect any device that produces TTL outputs to the Trigger In connector of the STG for example a switch For example you can use a trigger for remote controlling the STG if it is not within reach during an experiment It is also possible to set up advanced experiments where stimulation depends on preceding activities of the studied object The following picture shows a suggested circuit diagram for a switch used for remote controlling The resistor and capacitor work as a low pass filter on the TTL signal and are necessary to reduce ringing of the signal Suggested switch for triggering the STG 1 ko 3 3 V tob V 59 10 3 Stimulus Generator 4000 Series Manual Trigger Settings Triggering output channels Standard mode You can use up to four or eight separate trigger inputs for triggering up to four or eight output channels STG4004 or STG4008 respectively The trigger settings are saved in the currently active MC Stimulus II stm file Please assign the four triggers to the analog output channels in the Trigger Settings dialog box in the Edit menu Trigger Settings File Made Cont Made Standard Mult File Extended Multifile On C Off Settings Ch Che Chaa Ch45 rEh5b4 Che Lh 5 So S4 rHepeat Triggeri ie C i s C ie C e grs lee oo e e oo i elo v EE Joc ells lles lls
69. gher load resistance the current pulse shows a different kinetic behavior The rise time increases significantly about 100 us Please keep in mind that the resistance of the oscilloscope probe with 10 MQ is parallel to the 1 MQ load resistance That means that the maximum of the load resistance is reduced to 900 kO only 84 Analog Output Signals 11 6 Comparison of Output Signals in Current Mode To show the different behavior of the output signals in current mode there are some examples to get an idea how the stimulus generator will deliver the stimulus pulses The screen shots are from a standard oscilloscope triggered on the Sync Out pulse see Measuring output signals with an oscilloscope Example 1 Influence of the length of a current pulse 100 us versus 20 us with an amplitude of 1600 pA which is at the maximum capacity of the STG4000 series constant amplitude and load resistance Example 1a Current pulse with programmed 1600 UA for 100 us and 20 us interval and 10 kQ load resistance The pulse is biphasic and 1600 pA which is the limit of the maximum capacity of the STG 4000 series Green trace current output I pink trace TTL trigger output from the Sync Out 10 0V CA4 5 00V 10 20 85 Stimulus Generator 4000 Series Manual Example 1b Current pulse with programmed 1600 pA for 20 us and 20 us interval and 10 kQ load resistance The pulse is biphasic and 1600 A which is the limit of the maximum
70. he site is changed Damaged power cords should be replaced immediately and may never be reused Check the leads for damage Damaged leads should be replaced immediately and may never be reused Do not try to insert anything sharp or metallic into the vents or the case Liquids may cause short circuits or other damage Keep the device and the power cords always dry Do not handle it with wet hands Requirements for the installation Make sure that the device is not exposed to direct sunlight Do not place anything on top of the device and do not place it on top of another heat producing device Otherwise the device may overheat 3 1 First Use of the Stimulus Generator Welcome to the STG and MC Stimulus Product line overview Stimulus generators of the 4000 series are general purpose stimulators designed for a wide variety of applications both in vitro and in vivo Flexible and easy to use MC Stimulus software enables complex stimulus waveforms both current and voltage Waveforms designed in the program or imported from an external ASCII file are converted by the connected STGs into pulses which are sent to stimulating electrodes Stimulus isolation units SIU for each channel are included Thus no external SIUs are required A trigger input and output for TTL signals allows to synchronize the stimulus generator with other instruments Triggering of other devices via programmable TTL pulses Sync Out is possible as well as t
71. hen one of the channels that are assigned to the respective trigger input is still active Either the stimulus protocols on the triggered channels are stopped or restarted or the trigger event is ignored There is also a separate Sync Out output for each Trigger In trigger input You can assign the Sync Outs to the trigger inputs The default settings define that Sync Out 1 is active when Trigger In 1 is triggered and so on Please see Trigger Settings for more details STG status control Three status control LEDs light emitting diode are present on the front panel of the STG to give you a quick overview on its status POWER STG has been switched on PROG Stimulus file was downloaded to STG STG is now ready for operation SWEEP Stimulation is running that is at least one output channel is active MC Stimulus II status control The top part of the MC Stimulus II status control refers to the active instance of the MC Stimulus Il program instance and the STG that is controlled by this instance All information in this part is saved on the STG The information is updated in the MC Stimulus Il main window when the connection to the STG is changed or interrupted Status light Color Meaning Connected Green STG is connected and switched on The serial number serves as an ID for the controlled STG and is displayed on the left On the STG menu click Select Device to select another STG connected to a different USB port from the drop down l
72. hh mm s3 zzz Auto Sync for Autocreating Trigger Pulses If you want to synchronize other devices for example a MEA1060 BC amplifier or the MC Card with the stimulus generator you have to set up the digital output of the Sync Out channels in synchrony to the stimulus pulses This may sometimes be a bit tricky and time consuming especially for complex stimulus protocols The Auto Sync feature is intended for making the work with MC Stimulus more efficient Digital Sync Out TTL pulses synchronous to stimulus pulses are generated automatically by this feature Each time an analog channel generates a pulse that is its value is not equal to zero the logical state of the assigned Sync Out channel is set to HIGH 21 You can set the time offset of the TTL s rising edge before the stimulus pulse and of the TTL s falling edge after the pulse If you change the stimulus protocol corresponding Auto Sync pulses are updated on the fly This feature is especially recommended for controlling the MEA1060 BC amplifier with superior stimulus artifact suppression On the Edit menu click Auto Sync Settings The Auto Sync Settings dialog box appears Auto Sync Settings E Pre Time T Post Time 100 31 5 8 Stimulus Generator 4000 Series Manual Assign the analog stimulus channels that will be used as the basis for autocreating the protocols on the Sync Out channels 1 to 4 For example if you use only stimulus channel 1 select Ch1 und
73. i stm sTG Batch Signal View Window Efi uate Start Ctrl R E Sop Ctrl4T Communication with STG Download Ctrl 4D Downld and Start Ctrl A Disable Autocalibration Select Device If you have more than one STG connected to the same computer please click under STG in main menu Select Device In the Select Device dialog you can choose which STG you like to control from the currently active MC Stimulus Il program The serial number is printed on the STG s case The serial number of the currently active STG is displayed on the status bar of the currently active instance of the MC Stimulus Il program Djaja e 52 sesso fa remo recon t ropes t sweer III III Click Select Device under STG in main menu Select the serial number of the desired STG from the STG list You can open multiple instances of the MC Stimulus Il program and control multiple STGs at the same time Please make sure that you control each STG only by one MC Stimulus II instance You can also open only one MC Stimulus Il instance and control multiple STGs one after the other by the same instance Simply change the settings in the Select Device dialog and select the next STG when you have finished to program the first The STGs can be operated without a computer connection 14 Operating the STG 4 2 1 Operating the USB MEA32 STIM4 System The USB MEA32 STIM4 System has an integrated four channel current stimulus generator For operating this
74. if channel 1 has a stimulus with a length of 500 ms and channel 2 of 1000 ms and both are triggered by the same trigger event channel 1 waits 500 ms for the repeat until the output stream of channel 2 has been finished Note The Sync Out signal for the corresponding trigger is repeated as well each time the stimulus is repeated Hint You can repeat the complete stimulus protocol that is assigned to a trigger continuously by using the Continuous Mode setting Continuous mode can be activated in the main window or under Trigger Settings The STG has then to be stopped manually by the user You can use this feature regardless whether you use an external trigger input or not When you press the Start button this is considered as if all four trigger inputs were active When you use Continuous Mode the repeats entered in the Trigger Settings dialog box are disabled Cont Made Output Mode f On t Yoltage 26 01 2010 11 58 C Dif Current Trigger Settings File Mode Cont Mode E Standard Mult File Extended Muttifile On Off 1 Onthe STG menu click Trigger Settings 2 Enter the number of repeats for each trigger separately 0 repeats the output signals continuously 23 Stimulus Generator 4000 Series Manual In the following example the stimuli on output channels 1 3 and 6 that are all triggered by Trigger 1 are repeated three times after the trigger event each time accompanied by the Sync Out 1 outp
75. igger Settings File Mode Cont Mode C Standard C Multi File Extended Multifile C On amp pi Settings emen Femme T L4 00000100 C STG HODUMC STG4DDO OverWewsim ur L 8 0000iD00 CASTG MODOMC STG4DD0 Groups i L 18 00010000 CASTG MODOMC STG4DDO GrounRepesbsim i 32 DDi00000 CASTG HODOMC STG4DDO Grouphepes 2 im L9 foo wgn fe To arrange the created stimulation files you have to click on the grey square on the right side of each row A dialog will open and you can choose the file you want to place in this position The complete arrangement of the files can be saved as Extended Multi File configuration file with the command Save and load again with Load The configuration file for the Extended Multi File Mode has the extension mf2 When using the option Show all Patterns it would be hard to fill in files to each of the 256 positions You can shorten the procedure by editing an available Extended Multi File with a common editor 65 Stimulus Generator 4000 Series Manual Please make sure to use the correct format lox Datei Bearbeiten Format Ansicht Filenames Fi ledod c 4576 4 0005MC_sTG4 000_GroupRepeat stm Fille nl Filen 2z Filen s3 File n4 2c ssTG 4 00 MC sTG4O000 overview stm Fille n5 Filen 5z Filen ENEDDEES S164 Q OO0 MC sTGd OO0 GroupRepseat stm Fllegos Fi
76. imulus II file to the STG 3 Start the STG either manually by pressing the Start Stop button on the front panel with the software controls or with an external trigger Creating stimuli Stimuli are created user friendly by entering the desired pulses rectangular ramp or sine waveforms into a MC Stimulus worksheet Waveforms can be combined freely to create virtually any stimulus pattern Created stimuli are displayed in a WYSIWYG window Starting stimulation Note You need a computer connection only when you are using the software controls for starting and stopping the stimulus generator When you are using the STG interface or an external control trigger you can disconnect the stimulus generator from the computer and operate it independently But you cannot remove the STG from the power connection The memory is cleared when the STG has lost power After the download of the stimulus file on the STG you can start the stimulation There are several ways to start the STG MC Stimulus You can start the stimulation directly after the download with the command ll interface Download and Start I or also later with the command Start Lal You can stop stimulation with the command Stop Starting the STG by using the Start command is considered as activating all trigger inputs that is all channels are started and all Sync Out outputs are active Likewise all channels are stopped when you click Stop You can start the stimulat
77. ion after download T on up to eight trigger separately Please use the combined LED Start Stop buttons one SI Sei meer mer F F You can start and stop the stimulation manual whenever you want by clicking the corresponding LED Start Stop button The LED indicates the status Black means trigger off red means trigger on The first button on the left side belongs to trigger 1 the second button to trigger 2 and so on 11 Stimulus Generator 4000 Series Manual STG You can start the stimulation by pressing the Start Stop button on the front interface panel of the STG Press the button again to stop the stimulation Starting the STG by pressing the Start Stop button is considered as activating all four trigger inputs that is all channels are started and stopped and all four Sync Out outputs are active when the STG is started Pressing the Start Stop button stops the stimulation if at least one channel is still active in the moment of the trigger event indicated by the lighting SWEEP LED Pressing the button starts stimulation only if no channel is active dark SWEEP LED External You can start the stimulation on a trigger from an external device for example control a switch You can use up to eight trigger inputs that independently trigger the analog output channels that you have assigned to them The trigger works in a similar way as when you press the Start Stop button You can choose the action that will be taken w
78. is filled automatically In the following example the Autofill feature has been used to create a voltage step series from 20 mV to 500 mV with an increment of 20 mV Click any Value or Time column header to select a column The selected column is highlighted in black Right click and click Autofill Select one out of three commands and fill in the white text boxes Channel 1 Channel 2 Channel 3 Channel 4 Autofill Pulse value Unit time Unit value Unit 1 T recense 00 mwao us al v Bei 100 es NR 4 rectangular noo mM 40 us S rectangular 100 mw 4b s B frectanguer 100 mv 40 us x Sync 1 Start Stop Step Size C Start Value Ey Stop value i 3n zm zz 222222 i ra G Shep Size 1 na Hof Steps C 1 5 EIITI A ENHE IIHI O10 00 00 000 00 00 00 00 t hh mm ss zzz 30 5 7 Programming Stimulus Protocols The column is filled according to the specifications Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 Pulse value Unit time Unit value Unit time Unit value Unit time Unit Row rep Group r iv aedem o mwj us NE 3 rectangular o mo vs a frenoa o mwie e B EM rectangular o mv 4s us A e rectenguler o mw ps A rectangular O mv 40 e EW a So p mw 5 m 132 z 00 00 00 000 00 00 00 001 00 00 00 0072 t
79. is manual This documentation refers only to the STG hardware and MC Stimulus Il software For more information on the STG s DLL functions and programming of custom programs please read the separate documentation in the Documentation folder in the MC Stimulus Il program directory If you are using the STG for the first time please read the important safety advice before installing the hardware and software see Important Safety Advice where you will find important information about the installation and first steps The printed manual and Help are basically the same so it is up to you which one you will use The Help offers you the advantage of scrolling through the text in a non linear fashion picking up all information you need especially if you use the Index and the Search function If you are going to read larger text passages however you may prefer the printed manual The device and the software are part of an ongoing developmental process Please understand that the provided documentation is not always up to date Check also the MCS web site www multichannelsystems com from time to time for downloading up to date manuals and new software versions 1 2 1 3 Stimulus Generator 4000 Series Manual Terms of Use for MC Stimulus Il You are free to use MC Stimulus Il for its intended purpose You agree that you will not decompile reverse engineer or otherwise attempt to discover the source code of the software Limitation of
80. ist to switch STGs Programmed Green Stimulus file in voltage mode has been downloaded to STG STG is now ready for operation File name and path of the stm file that was downloaded onto the STG are displayed on the right Orange Stimulus file in current mode has been downloaded to STG STG is now ready for operation File name and path of the stm file that was downloaded onto the STG are displayed on the right Sweep Red Indicates the active trigger up to eight Each stimulation channel can be assigned to any trigger If all eight channels are assigned to trigger one for example only one sweep indicator will light up even though all stimulation channels can be active See also Triggering Stimulation 12 Operating the STG STG and Stimulus Protocol Information Information on the STG hardware Serial number STG status light Sweep LED Start Stop Voltage and current range p MC_Stimulus II MC_Stimulus_Main_Menu_File stm File Edit STG Batch Signal View Window Settings Help Dag siel amp 2 ay Tp n ste 80002 Disconnect m Programmed ES Sweep IL _J_ _J_ _J_Ivotage Range 8000 nv Current Range 1600 pA MC_Stimulus_Main_Me E MC Stimulus II in Menu File stm Channel 1 Last Download Cont Mode r Output Mode r Download Channel Download Sync Comment Signal Length 9 920 ms 80002 On Voltage lv1lv2iv3lv4di ivilv2iv3iv 4 max Amplitude 2 500 V 22 01 2010 14 14 min Amplitude 3 000 0
81. istance is reduced to 900 kO only The long pulse is biphasic 50 uA Green trace current output 1 pink trace TTL trigger output from the Sync Out H1 00ms A Cha 1 60 V 20 0V weh4d 5 00 V iB 10 20 Result of Example 4 The current output signal is delivered negligible modified when the duration of the signal and the duration of the interval is long for example 1 ms 90 Analog Output Signals 11 7 Capacitive Behavior of Stimulation Electrodes Regarding the generally used stimulus pulses stimulating electrodes behave as plate capacitors They need some time to discharge themselves after stimulation As a result artifacts interfere with the recording and electrodes deteriorate over time due to electrolysis This effect takes place especially in current mode because the current cannot flow back to the stimulus generator due to the high output resistance in current mode and thus is kept in the electrode A current source can constantly produce small currents even though the current is set to zero This might damage the connected electrodes over time To avoid this problem the STG4000 series is equipped with switches that automatically disconnect the current outputs from the current source 1 s after each programmed current stimulus For the next stimulus the current source is automatically reconnected To reduce the effect described above in current mode you should use biphasic pulses for stimulation The stimulus signal shoul
82. ither rectangular ramp or sine Enter the desired voltage steps and a time length for each step Channel 1 Channel 2 Channel 3 Channel 4 Pulse __ value Unit time unit rectangular 1000 mv 0 T 6 rectangular 19 5 4 Stimulus Generator 4000 Series Manual Pulse Types Three basic pulse types are available in MC Stimulus Il You can create virtually any pulse by combining different types Rectangular Ramp Sine wave Note The time resolution is 20 ps When you enter time steps that are not a multiple of 20 us the length of the step is internally adjusted rounded arithmetically that is 249 us would be rounded down to 240 us 250 would be rounded up to 260 us to a valid value The minimum voltage resolution is 1 mV the current resolution is 200 nA Please regard also the minimum and maximum output voltage current of your STG version If you enter voltage or current values outside the STG s limit the invalid value will be changed automatically when you download the protocol Cells with changed values will be highlighted in yellow Note If you work near the resolution limits of the STG the output pulses may differ from the programmed pulses In this case you should check the output with an oscilloscope Please see also the Analog Output Signals chapter z Channel 3 CF Please note that the memory of the STGs of 4000 series is limited 64 MB You will get an error message if the memory is fu
83. kind of STG please select the device in Select Device dialog via serial number To select stimulation channels open the dialog Select Stimulation Channels in Edit menu Select Stimulation Channels X Assign Ehe 4 stimulation channels to electrodes orf 51 S2 53 54 55 56 Channeli T fw fF FO rr rE Channel Nw PF fF T T FL off 5 58 5S9 6510 511 Sle Channel 3 I Iw fF rr r chamel4 T YF TV FT F fw Select Layout 1 x 12 It is possible to assign stimulation channel 1 and 2 to stimulation electrodes S1 to S6 It is not possible to select the same electrode for both channels Stimulation electrodes S7 to S12 have to be assigned to channel 3 and 4 If you do not need four channels for stimulation click check box channel off For programming the stimulation channels please use MC Stimulus II as usual 15 5 1 5 2 Programming Stimulus Protocols MC Stimulus Il Sample Files for STG 4000 Series These sample files are intended for demonstrating basic features of the Stimulus Generator STG 4000 series for basic applications with the MEA System If you are using an MEA System please see also the MC Rack tutorial in the MC Rack help or the separate PDF on the installation volume The sample files are automatically installed to the Examples folder in the MC Stimulus Il program directory Please note The demo files were kindly provided by STG users MCS does not guarantee that these files will be appropri
84. le 21 2 1 Connect the digital IN OUT connector to a TTL signal source 2 Ground digital inputs manually which are not used 3 Set up your own stimulus protocols either nine stimulus patterns or up to 256 patterns and save the files 4 Onthe Edit menu click Trigger Settings 5 Select the option Extended Multi File 6 Optionally select the check box Show all Patterns 7 Define the desired files to each of the positions or load a configuration file mf2 for the Extended Multi File mode 8 Close the dialog by clicking OK 9 All selected files are opened automatically 10 Check all stimulus protocols before download Please make sure that you have selected all necessary channels under Download Channel for each file separately 11 Download the files onto the STG by clicking Download on the STG menu 12 The stimulation as specified in the selected MC Stimulus II files will now be triggered by trigger input 1 66 Synchronizing Events 10 6 Batch Mode You can download and run several stimulus files one after the other in the Batch mode The first file in the list is downloaded and started directly after the download or started manually with the software or hardware controls or started on a trigger After the first file has been run the next file in the list is downloaded and so on until the last file in the batch list has been run Triggering a sequence of stimulation protocols The Batch mode is use
85. ll For the digital Sync channels only TTL pulses with bit patterns of 0 LOW and 1 HIGH can be programmed A logical state of 1 HIGH results in a 3 3 V output signal and a logical state of O LOW results in a O V output Please note that the digital output is sent about 20 ps faster than the analog output Please see also the Triggering Stimulation chapter Rectangular pulse type Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 walue Unit time Unit value Unit time Unit value Unit time Unit Row repe 1o mw 2p me xm mw ims o mvi ms 2 z i oe cE eS r M k rT A an he f Dior au 00 000 Dor ac por Oa Dur ad dor aad Dur ad mi aeu Dur ad du dau t hh mm ss zzz EA amp amp rv cm C che M Ch3 M ChA M Chs M che D kom mmf Z7 Ready Enter each voltage current level and the duration of the pulse The voltage current jumps directly to the specified level In one row you can enter up to three voltage current levels Use the next row for programming more levels 20 Programming Stimulus Protocols Ramp pulse type Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 _ Pulse value Unit time Unit value Unit time Unit value Unit time Un Q2 cac FETTE Lt 00 00 00 200 00 00 00 220 00 00 00 240 00 00 00 260 00 00 00 2860 t hh mm
86. log box with the Repeat option the STG abruptly stops after it has generated the waveform only once If another STG is connected to the computer to run the same protocol the stimulator has no problems on repeating the pulse Possible causes If you change the stimulus protocol after the download even if only the number of repeats were changed you need to download the modified stimulus protocol to the STG Otherwise the changes will not be applied If another STG is connected to the computer you will have to download the stimulus protocol including the changes to be able to stimulate at all Select Download or Download and Start on the STG menu or on the toolbar after you made any changes to the stimulus protocol in the MC_Stimulus program Internal Calibration Signals The STG emits sometimes autonomous high frequency signals Cause During the first ten minutes after starting the STG the device emits every minute high frequency signals small bursts for internal offset calibration of the current outputs in a range of 10 mV After warming up the STG the calibration signals later will be delivered half hourly The calibration is interrupted during stimulation High sensitive measuring equipments will monitor these calibration signals Biological signals are usually slower than the high frequency bursts and will not interfere with the calibration signals When monitoring the experimental signals with MC_Rack software
87. ltage in microvolts uV or current in microamper pA This file format is used by MC Rack ASCII export You can directly import the MC Rack export files into MC Stimulus II time value You can use this format with or without header Example The following picture shows a file of format type 6 without header T PelecOOO1 z9 nV O 000040 B o521557 0 000080 lo 7500 0 000120 le 708333 0 000160 0 054166 0 000200 7 ll83333 0 000z 40 S T4H1857 0 000280 15 104167 D 000320 13 854155 0 0003 60 le 500000 0 000400 1 875000 42 ASCII Import Export The following picture shows a file of format type 6 with header Multi Channel systems MC Stimulus ASCII import Verzion 1 10 channels amp 8 output mode format 6 channel 1 time value HV O 000040 2 541667 O 000080 2 167500 O 000120 12 708333 0 000160 6 054166 0 000200 7 O8S535 O 000240 5 729167 0 000 a0 15 104167 0 000320 13 8554166 0 000360 i 500000 0 000200 1 675000 Using the format type 6 without header only the first channel of the MC_Rack raw data file can be imported Using the format type 6 with header you choose how many channels are imported in channels line 3 The output mode in line 4 has to be voltage You choose which channel should be imported first in channels line 6 You can add the channels you also want to import one after the other in any order The value microvolt is normally used for data recording in voltage mode of MC
88. make sure that you have selected all necessary channels under Download Channel for each file separately Download the files onto the STG by clicking Download on the STG menu The stimulation as specified in the selected MC_Stimulus files will now be triggered by the selected trigger inputs Trigger In 14 Triggered action when channel s still active You can define what action should be taken when any of the channels assigned to the trigger are still active In addition you can also define what action should be taken when any of the channels F not assigned to the trigger event are still active Please click the expand button to show the advanced options Stop A following trigger input stops the stimulation on the assigned channels if at least one channel that has been assigned to this particular trigger is still active in the moment of the trigger event The trigger event starts stimulation only if no channel that has been assigned to this trigger is active Restart The stimulus protocols on all channels assigned to this trigger are started at the trigger event regardless of the activity on these channels Ignore The trigger event is ignored when any channel assigned to this trigger is active The trigger event starts stimulation only if no channel that has been assigned to this trigger is active Example File A is assigned to trigger 1 and file B to trigger 2 Consider the following two situations Trigger 1 is activated
89. me Unit Row r a frectancuer fe mv 20 js e mvoo js mv p 2 rectancuiar 6 mv 20 us 5 mv 20 us fm p 3 rctenue 6 mv 20 jus e mvo us mv p meena o ENS 5 c Hio 5 EM 0 5 rectangular 1 mi 20 5 mw 20 m Ls 4 k 182 7 ov rr oo T f ct qq at N Duce 0000500 0 001 000 0 001 500 0 002000 0 002500 0003000 t ASCII import example Stimulating with biopotential waveforms Shown is a single spike recorded from an organotypic hippocampal culture The spike waveform was exported as ASCII from MC Rack and imported into the MC Stimulus Il program Warning All previous records in the active MC Stimulus II file stm file are overwritten Please create a new file and import the data into an empty worksheet to avoid data loss On the File menu click Import ASCII File The Open dialog box appears Select an appropriate ASCII file and click Open The stimuli from the ASCII file are loaded into the active MC Stimulus worksheet All previous records are overwritten Channels that are not present in the ASCII file are empty You can now edit and download the stimulus file as usual If the file does not fulfill the requirements of a stimulus file an error message will show up and tell you in which line the first error occurs See Supported File Format for more information about supported ASCII files 35 6 2 6 3 Stimulus Generator 4000 Series Manual Exporting Files On the File m
90. n arrange the windows with the commands on the Window menu Cascade Use this command to arrange multiple opened windows in an overlapped fashion The windows are resized to the standard size Tile Use this command to arrange the windows so that they are not overlapping Arrange Icons Use this command to line up minimized windows You can resize the windows with your mouse Click an empty area on the toolbar and then drag the toolbar to a new position Menu Bar You will find most software features in the pull down menus of the main window Each menu displays a list of commands Commands that are not available at the moment appear shaded Use the File menu to create new files save files import ASCII files and print the stimulus worksheet Edit menu for general editing features like copy and paste rows remove the last entered value Undo remove and insert rows into the active worksheet You can also right click on a row and select these commands Click Clear Signal to delete all values in the active worksheet You can also specify the output ranges according to the connected STG on this menu Please see the technical specifications of your STG The STG menu contains all commands directly relating to the hardware for example download commands start and stop the STG and trigger configurations You can open start and stop a batch run of several files on the Batch menu Signal menu for nice editing features like editing
91. ng mode In Download mode you can control the STG either by the MC Stimulus Il or by a custom program Stimulus protocols are created in MC Stimulus II or the custom program complete protocols are downloaded onto the STG and can after the download be executed without a computer connection MC Stimulus II is downward compatible to MC Stimulus for controlling the 1000 series that means you can load stimulus protocols generated by MC Stimulus into MC Stimulus II but not the other way round In the Streaming mode you can control the STG either by the Stimulus Streamer or by a custom program If you use the Stimulus Streamer you can load stimulus protocols in Mono MP3 or binary file format of unlimited size that were generated by a custom application and are then sent continuously from the computer to the STG in small data packets If you use a custom program for controlling the STG this program can generate and send the stimulation protocols in parallel from the computer to the STG Custom applications for the Download or the Streaming mode can be programmed in Matlab Labview C C and Visual Basic This documentation refers only to the STG hardware and MC Stimulus Il software For more information on the STG s DLL functions and programming of custom software please read the separate documentation of the Documentation folder in the MC Stimulus II program directory First Use of the Stimulus Generator MC Stimulus Il Stimuli are crea
92. nload new stimulation paradigms to individual channels without interrupting the ongoing stimulation on other channels See chapter Downloading Stimulus Files for more information It is mandatory that all stimulation and Sync Out channels that are associated with one amplifier are assigned to the same trigger in the Trigger Settings menu For example if four MEA1060 BC amplifiers are connected to one STG4008 usually two stimulation channels and one Sync Out are associated to each amplifier In that case the trigger settings should be as shown Settings Ch Che Cha Ch4 ChS ChE Ch Che 5 a2 53 54 Tmagerl i C C b C C C E amp C Tiggere 7 C s O C C C a O C C ovs fa e e le to flo lle e e lo lo Tnggert 7 C C C C C E C Tigger 7 C a C C C C L C C C co edle lles e w e i t Tigger 7 C C C C C C C C C E C cec f e e we o ls e 9 ls wt Amplifier A is connected to stimulation channels 1 and 2 and Sync Out 1 Amplifier B is connected to stimulation channels 3 and 4 and Sync Out 2 and so on All channels connected to one amplifier are assigned to the same trigger 51 Stimulus Generator 4000 Series Manual To download a stimulation paradigm to individual channels assigned to one amplifier only select all channels connected to this amplifier in the Download Channel check box 1 and 2 in this example and Sync Out 1 in Download Sync check
93. not use any other trigger inputs Splitting up stimulation protocols to circumvent the memory limitation If the option Start STG after Batch Download on the Settings menu is selected the STG starts each file directly after the download which is only useful in the unlikely case that you need to circumvent the memory limitation of the STG Split up your pulse protocol into several stimulus files and run them in Batch mode Setting up a batch file You have to set up a batch file stb first which lists the full path of all files that you like to download as a batch The batch file has to be exactly in the following format This example is a batch list with three different pulse protocols Replace the file paths with the correct path of the files of your choice Multi Channel Systems MCS MC_Stimulus Batch Control File Version 1 00 c Program Files Multi Channel Systems MC_Stimulus TI BatchExample Baseline stm c Program Files Multi Channel Systems MC_Stimulus IINBatchExampleNTetanus stm c Program Files Multi Channel SystemsMMC Stimulus IINBatchExampleNTestResponse stm 67 Stimulus Generator 4000 Series Manual Operating the STG in Batch mode Important You can operate the STG in Batch mode only with a valid computer connection A file in the batch list is only downloaded after the preceding file has been run On the Batch menu click Open to open a batch file The Open dialog box appears Browse your folders and select a
94. nstalled software does not work properly Double click Setup exe on the installation volume The installation assistant will show up and guide you through the installation procedure Follow the instructions of the installation assistant Please select the appropriate maximum number of analog output channels according to the connected STG If you have different STGs in use choose the highest number For the devices with internal stimulation STG3008 FA and MEA32 STIM4 separate manuals are available STG Settings X Choose your Device C sTG4002 C sTG2004 STG4004 C sTG2008 sSTG4008 STG3008 FA MEA32 STIM4 Channel Configuration Analog Channels s Syncout Channels 6 T Trigger Inputs caa When you have finished the installation of the MC Stimulus II software and the STG driver you can now operate the STG 10 4 Operating the STG 4 1 Operation Overview Warning Make sure that you do not come in contact with the cables or the connectors N of the STG after you have started the STG The high voltage and power can lead tO Injuries p Warning Do not start the STG if you are unsure about the channel configuration or the nature of the downloaded file 1 Setup your experiment 2 Switch the STG on Note It is recommended to start the STG ten minutes before use to allow warming up 1 Setup your stimuli with the MC Stimulus Il program 2 Send the stimulus sequence from a MC St
95. ocol is longer than the gating trigger stimulation will stop when the gating trigger ends For example if you want to produce a repeated rectangular pulses that last as long as the gating trigger is HIGH you can do the following program a single rectangular pulse and activate the continuous mode The pulses will start as soon as the gating trigger becomes HIGH and stop as soon as the gating trigger becomes LOW If the voltage of the stimulation channel is different from 0 at the time the gating trigger ends the channel will be switched off immediately If you stop the stimulating protocol manually during the gating trigger in HIGH stimulation is stopped but immediately restarted and goes on until the gating trigger ends Warning Once selected this setting will become the default setting for all opened files until unselected So if you observe unexpected behaviour when triggering the STG check the Trigger settings Single Trigger Important This feature is available only with STG4000 series The MC Stimulus Il version 3 2 3 or later with firmware update must be installed Additional to the general trigger states running and stopped each trigger can now be set up in a new state called armed That means the trigger is ready to start the stimulus protocol but waiting for an external signal from the rear panel trigger inputs to execute the stimulus protocol one time only In armed state the Sweep LED on the front panel of the S
96. of output channels For example if you have installed MC Stimulus II for 4 channels the Sync Out channel is channel number 5 if you have installed MC Stimulus II for 8 channels the Sync Out channel is channel number 9 The data follows after the channel specification line three format types are available All format types follow the MC Stimulus Il worksheet s structure of rows and columns See the description of the worksheet for more details Columns are separated by blanks or tabs The first line has to contain the column headers The following lines contain the voltage current and time values Rows lines are separated by a carriage return or line feed CR LF This is the Windows standard control character inserted when you press the ENTER key The voltage unit is mV and the current unit is pA Please make sure that all values are in the ranges specified for the STG you use For the Sync Out channel only values of 0 and 1 are allowed The time unit is pis Time values can be in the range from 20 to 18 000 000 000 20 ys to 5 h Time base of the STG is 20 ps so only times that are multiples of 20 ps are valid Note Please make sure that there are no additional tabs or characters which would lead to an error message Show tabs and spaces in your text editor and check the file before importing it 37 Stimulus Generator 4000 Series Manual Format type 1 This is a basic format with two columns of Value and Time each without Repeat f
97. ogrammed with MC Stimulus II of 300 us length for a biphasic pulse of 200 us length See also the MEA Select help or the MEA Amplifier with Blanking Circuit manual for more information Channel 1 Channel 2 Channel 3 Channel 4 Channel 5 Channel 6 Channel 7 Channel 8 SYT i ee e LETS ESI ES ETT E 1 rectenauer p mv ms 1000 mV 100 000 os on mms 0 m wm 1 Em rectangular mv Hs mi mz You can use the Auto Sync feature for autocreating the Sync Out pulses without entering values manually Please see Auto Sync for Autocreating Synchronous Sync Out Pulses Triggering Stimulation Trigger In The external trigger input has to be a TTL signal of at least 40 10 ps length TTL pulses shorter than that may not be recognized by the stimulus generator A TTL pulse is defined as a digital signal for communication between two devices A voltage between 0 V and 0 8 V is considered as a logical state of 0 LOW and a voltage between 2 V and 3 3 V means a 1 HIGH It is the same kind of signal as the output signal of the Sync Out output of the STG The rising edge of the HIGH trigger input starts and stops the STG The time lag between the rising edge of the TTL input and the stimulus output is 70 10 ps The minimum distance between two trigger inputs is 100 ps resulting in a maximum trigger frequency of 10 kHz If a higher frequency is used the stimulus generator is likely to miss trigger in
98. onding Auto Sync pulse Pre Time O us Post Time 100 us on Sync Out 1 If you later choose to edit the Sync Out worksheets of the active file manually you can deselect the Auto Sync feature on the Settings menu Any information in the Sync Out worksheets that was there before the Auto Sync option was enabled will be restored and all worksheets will be available for manual editing Adjusting the Stimulus Intensity Level After having programmed the stimulus protocol you can either download the protocol as is or you can adjust the stimulus intensity level that is choose a percentage from 0 to 200 percent of the programmed amplitudes independently for each of the up to 8 channels You can adjust the level offline and download the information onto the stimulus generator manually afterward or you can adjust the level on the fly without stopping the running stimulation This feature is especially convenient for testing out thresholds that evoke a biological response Note The error messages that are displayed when the stimulus amplitude is not set to zero at the end of the protocol or when the time length of stimulus protocols is different on individual channels can present a nuisance when adjusting the level on the fly These error messages can be switched off by deselecting the option Enable Compiler Warnings on the Settings menu On the Edit menu click Amplitude The Amplitude dialog box opens You can drag and drop it anywhere yo
99. or ASCII import files with or without header Please choose the factor of amplification in Amplification This is very helpful for example for importing data recorded in microvolt uV and now needed in millivolt mV That means the factor is 1000 or for higher stimulation signals may be 10000 Please see chapter Format Type 6 above Important MC Stimulus Il uses the header of an ASCII import file if a header is available If no header is available MC Stimulus Il applies the settings of the dialog ASCII Import Setting 44 7 1 7 2 7 3 Stimulus Display Selecting Channels To enlarge the display size of the channels we suggest that you show only those channels you are currently working with Note This will affect only the display It has no effect on the download of signals To select or deselect channels for downloading click the Download Channel or Download Sync check boxes in the main window of the program Clear the check box of all channels that are not in use All channels that are not selected are removed from the display a a e a JJ a SG fae fae a aa NIE NIE NIE NE NAE NI ae a 00 00 00 000 00 00 00 200 00 00 00 400 00 00 00 600 00 00 00 800 00 00 01 000 00 00 01 200 00 00 01 400 hh mm ss zzz Ch4 ChS Ch6 Ch Ch8 Sync2 Sync3 Sync4 Syncb J Sync Sync Sync8 Display Settings You can toggle between a decimal time scale in seconds floating point numbers and a
100. oring stimulating electrodes is needed The current that is delivered by the first electrode flows directly back to the second electrode so that the surrounding tissue is not affected See illustration Voltage mode Bipolar stimulation In the following example you see a signal in the MC Stimulus II display and the resulting output signals for the different connection options viewed in a MC Rack Data Display Programmed stimulus protocol viewed in MC Stimulus Programmed signal S a T T a fn 0 0000 0 1000 0 2000 03000 0 4000 0 5000 0 6000 t s 70 Analog Output Signals STG output signals imi Display 1 358610sec Xrdxis 1000 ms Bth sweep Y Axis t2 s 100 150 200 250 300 350 U to GND 14 1 6 14 1 1 0 8 0 6 0 4 D D 4 B E 0 8 1 l 14 16 18 2 386 987 sec wis 1000 ms Sth sweep Ys 2 V 30 23 50 100 150 200 250 300 350 400 450 500 550 PG sx Seeks xc ee er ee eer ee IEE gii ph IET gii pp EL gd gag EL gi ggg 329 659 sec XAxis 1000 ms 00 p3 ron 7th sweep Ys 2 V vj ZU q S 50 100 150 200 250 300 350 400 450 500 550 600 i2 I T Ph 14 1 1 0 8 0 6 0 4 n 0 4 n D 1 1 1 4 T 1 2 71 Stimulus Generator 4000 Series Manual Screen shot from a standard oscilloscope triggered on the Sync Out pulse see
101. ouse pointer becomes a hand zou 100 00 00 00 000 DD DIEI While holding down the mouse button move the axis to the left and right or up and down OR You can use the keyboard Press LEFT ARROW or DOWN ARROW to move the axis to the left down RIGHT ARROW or UP ARROW to move the axis to the right up OR Use the PAGE UP and PAGE DOWN keys for a faster scrolling larger steps PAGE DOWN to move to the left down PAGE UP to move to the right up 46 Stimulus Display Zooming the display Tet 1 Click to enable the scaling 2 2 Click the axis you like to scale The mouse pointer becomes an arrow 10 Taamannuasuuuansanuauusa 00 00 00 000 00 00 3 While holding down the mouse button move the mouse up right to zoom the axis in and move the mouse down left to zoom the axis out OR You can use the keyboard Press LEFT ARROW or DOWN ARROW to zoom the axis out RIGHT ARROW or UP ARROW to zoom the axis in OR Use the PAGE UP and PAGE DOWN keys for a faster scrolling larger steps PAGE DOWN to zoom out PAGE UP to zoom in Size to fit a Click amp to size the display to fit The ranges are automatically adjusted to fit the maximum amplitude of each stimulus The range of the time axis will be set to fit the longest signal 47 8 1 8 1 1 Downloading Stimuli Downloading Stimulus Files After having set up the stimulus file you can download the file to the connected STG Yo
102. puts See the illustration below for more details on the timing Please note that the digital Sync Out output is about 15 ps faster than the analog outputs See Digital Output Signal Sync Out for more information You can either trigger separate output channels Standard mode or separate MC Stimulus II files Multi File mode There is also one separate Sync Out output for each trigger input You can assign the Sync Outs to the trigger inputs in the Trigger Settings dialog box The default settings are that Sync Out 1 is active when Trigger In 1 is triggered Sync Out 2 is active when Trigger In 2 is active and so on See also Digital Output Signals for more information on the Sync Out channels The Sweep status display LED Start Stop buttons in the main window 5 Weep s jn je t pn n E mE E mer of the program shows active sweeps that were started by the triggers That means the status lamp that corresponds to trigger 1 lights red as long as any channel triggered by trigger 1 is active Please note that the number of the analog output channel is irrelevant the Sweep status lamp refers only to the trigger number Example Assume that channel 4 is assigned to trigger 1 When channel 4 is active the first Sweep lamp lights red because the first Sweep lamp refers to trigger 1 58 Synchronizing Events You can start and stop the trigger manually at any time using the LED Start Stop buttons Timing limits Stimulus output Trig
103. r on to continue your experiment Save as Use this command to save your file under a new name for example if you like to use a file as a template for a new file On the File menu click Save As Browse your folders and enter a file name Click Save The current worksheet is now saved under the new name You may open the file using MC Stimulus II later on to continue your experiment Import ASCII File Please see the chapter ASCII Import Export Export ASCII File Please see chapter ASCII Import Export Printing a channel On the File menu click Print Setup to select a printer paper format and so on Click Print Preview to preview the print output Click Print amp to print out the active channel of the active worksheet 59 Stimulus Generator 4000 Series Manual 9 6 Settings Menu Enable Compiler Warnings Cancel this option to disable the message boxes displaying compiler warnings during stimulus download The two error messages that can be disabled are warning signal length shorter than the maximum and warning non zero last amplitude value They may present a nuisance if you download multiple stimulus files in batch mode or if you want to adjust the stimulus intensity level on the fly On the Settings menu select or deselect Enable Compiler Warnings 56 10 10 1 Synchronizing Events Digital Output Signals Sync Out The output of the digital Sync Out outputs is a 3 3 V TTL signal
104. re to read the following advice prior to install or to use the device and the software If you do not fulfill all requirements stated below this may lead to malfunctions or breakage of connected hardware or even fatal injuries Warning Obey always the rules of local regulations and laws Only qualified personnel should be allowed to perform laboratory work Work according to good laboratory practice to obtain best results and to minimize risks The product has been built to the state of the art and in accordance with recognized safety engineering rules The device may only be used for its intended purpose be used when in a perfect condition Improper use could lead to serious even fatal injuries to the user or third parties and damage to the device itself or other material damage Warning The device and the software are not intended for medical uses and must not be used on humans Malfunctions which could impair safety should be rectified immediately High Voltage Electrical cords must be properly laid and installed The length and quality of the cords must be in accordance with local provisions Only qualified technicians may work on the electrical system It is essential that the accident prevention regulations and those of the employers liability associations are observed Each time before starting up make sure that the mains supply agrees with the specifications of the product Check the power cord for damage each time t
105. riggering of the STG by external devices via the Trigger In input For example you can synchronize stimulation and recording with a TTL signal sent from the Sync Out output of the STG to the data acquisition system for example the MC Card Stimulus generators are available with two four or eight completely independent stimulation channels STG4002 STG4004 STG4008 Each channel is freely programmable combination of channels to achieve complex stimulation patterns is not necessary Stimulus Generator 4000 Series Manual 4000 Series ane shonin mulik en STARTISTOF ichannel multi ystems e POWER f 4 z v nm PRO gTARTSTOP WEEP GND 5 yel multi tems 2 power f proc f O aeTOr TAR rsTO g gweer 4 The real time stimulus generator of the 4000 series is an advanced version and the world s first STG that is able to send continuous pulse streams to stimulating electrodes In addition you can trigger all up to 8 channels separately by external TTL pulses 2 trigger inputs Trigger In and 2 trigger outputs Sync Out are shared by 2 channels in the 2 channel version 4 trigger inputs Trigger In and 4 trigger outputs Sync Out are shared by 4 channels in the 4 channel version and 8 trigger inputs Trigger In and 8 trigger outputs Sync Out are shared by 8 channels in the 8 channel version The STG 4000 series has two operating modes Download mode and Streami
106. rrent Mode The green trace shows the current output I with a load resistance of 10 kQ and the pink trace shows the TTL output from the Sync Out that was used for triggering the oscilloscope Rise time with programmed 5 V and 500 pA pulses 10 kQ load resistance Rise time 10 99 90 3 us H4 00us A Chd J 160V 2 00V Ch4 5 00 V ij 10 20 Result of Example 2 The kinetics and the rise time of the 5 V voltage pulse are comparable to the 500 mV pulse The approximate 3 us rise time of the 500 pA current pulse is not higher than the rise time of the 50 pA pulse 82 Analog Output Signals Example 3a Current Mode The red trace shows the current output I with a load resistance of 100 kQ and the pink trace shows the TTL output from the Sync Out that was used for triggering the oscilloscope Rise time with programmed 5 V and 500 UA pulses 100 kQ load resistance Rise time 10 90 96 12 us 730 0 V 3 EATE A Ch Y 160v zy 5 00 V B 10 20 83 Stimulus Generator 4000 Series Manual Example 3b Current Mode The green trace shows the current output I with a load resistance of 1 MQ and the pink trace shows the TTL output from the Sync Out that was used for triggering the oscilloscope Rise time with programmed 5 V and 500 pA pulses 1 MQ load resistance Rise time 10 90 120 us l gl i ME 20 0V Ch4 5 00V j j 10 20 Result of Example 3 With a hi
107. rted with the first file after the last file in the batch has been finished 68 11 Analog Output Signals 11 1 Output Modes The stimulus generator operates in voltage and current mode Choose voltage or current mode in the main window of MC Stimulus Il Please make sure that you use the appropriate stimulation mode for your experiment The difference between current and voltage controlled stimulation is described in the following In voltage mode the voltage level is held constant and the current output depends on the electrode resistance the higher the electrode resistance the lower is the output current In current mode the stimulus generator guarantees a stable current output independent of the electrode resistance where the voltage is defined by Ohm s law The higher the electrode resistance the higher is the output voltage Therefore the kinetic properties of the stimulus output is affected by the load resistance which is usually the electrode resistance If the resistance is too high the output current is limited by the compliance voltage that is too high resistances will result in a clipping of the output signal Please consider the time and voltage current resolution of the stimulus generator The minimum pulse has an amplitude of 1 mV in voltage mode or 200 nA in current mode supported by MC Stimulus II the firmware DLL supports a 14 bit resolution which means 200 nA for an output current range of 0 8 mA and a
108. rts downloaded stimulation file only available if STG has active computer connection Stop Stops downloaded stimulation file only available if STG has active computer connection eye Se EEEn o Shortcut Keys Shortcut keys offer you another way to accomplish common tasks Using a shortcut key usually consists of pressing and holding one key while pressing a second key See the list of shortcut keys available in the software Shortcut key CTRL C Copy Row CTRL D Download CTRL A Download and Start CTRL N Create New File CTRL O Open File CTRL P Print CTRL R Start CTRL S Save file CTRL T Stop CTRL V Paste Row CTRL X Cut Row CTRL Z Undo 54 9 5 General Software Features Hint Shortcut key combinations are also listed on the toolbar menus For example the File menu shows that the shortcut key for Save is CTRL S File Menu You can save MC_Stimulus II files for later use This is very convenient if you have various similar experimental setups or if you like to repeat an experiment later Open a file You may open previously saved files for similar experimental setups On the File menu or on the toolbar click Open u Browse your folders and choose the desired file You may only open MC Stimulus II files stm Click Open The selected file opens Save a file On the File menu or on the toolbar click Save Bl The currently active worksheet is now saved You may open the file using MC Stimulus II late
109. s Smei Synce synca Putse value Unit time uni 1 rectanguiar mv us Increment selected values 2 jeden Amv 0s oe o Cancel B frectarguiar i mv us 4_ rectanguiar mV JHS Type of Increment s enm fl my B jedes mv us Ni Keep Zero C Absolute value Inn EIN 3H zin rd TT THE TT 00 ad ad pao Dr an ad 01 t hh mm ss zzz All values in the selected column are modified by the selected factor In the example 100 is added to all values resulting in a total of 100 because the initial value has been O Channel 5yr J Puse E uni ime Unit vate une time Unit value unt time unit Row rep Group r ME MV 5 mv gs 2 recense M s mv us s rectangular RISE E 4 rectangular MEME rectangular MEN IB rectangular MEME n TT unn mu 5 r Hn Dor ad ad dao Dr an ad du1 t hh mm ss zzz 27 Stimulus Generator 4000 Series Manual Creating time or voltage current increments With this option you enter a base value for modification of all values in the selected column The first row is modified by the base value The second row is modified by the double value the third row by thrice the base value and so on With this feature you can easily set up time or voltage increments Under Type of Increment select Count Enter the desired start value Candi channel2 ch
110. s per phase Inter Stimulus Interval 24 ms ASCII Import Biopotential stm This file is not intended for a real experiment but demonstrates the ASCII import feature Shown is a single spike recorded from an organotypic hippocampal culture with the MEA System The spike waveform was exported as ASCII from MC Rack and imported into the MC Stimulus II program The imported file is ASCII Import Biopotential dat General Demo Files Demo Waveforms stm This file is not intended for a real experiment but for demonstrating the capabilities of the stimulus generator The file includes stimuli with increasing inter stimulus interval and with increasing amplitude ramps and sine waveforms 18 5 3 Programming Stimulus Protocols MC Stimulus Il Worksheet You can freely combine any pulse types Use a separate row for each pulse type Hint Several tools are provided for editing whole columns at once or autocreating entries Select the desired output type first Click either Voltage or Current under Output Mode on the file window of MC_Stimulus Output Mode Click a tab to select an output channel The according channel worksheet is brought to front Channel 1 2L Channel 3 Channel 4 Puise value tiit time unit value un aeta mvs 2 rectangular mw hs a rectang mw s a rectang mw s Seded mw s B rectang m hs Now you can enter the pulses into the worksheet Select a Pulse type e
111. sssse 25 lOSOTUTIDI ies dentes ti o eL 20 108 ise TITIO ssa ices ii senaicta ned estime Gases tirana 77 time offset for Auto Sync 31 Timing blanking of amplifier 57 HSE TING ono diss acidic Poo d cdd deos 77 Toolbar PI OSIULOD Mer ES 53 Trigger blanking signal 57 COMMOCIONS eR 8 CONTINUOUS mode 22 60 dice c 63 or lC 22 60 gating trigger eus 22 60 1g 81E 1 spec mE TU E ere meet oe eee 58 LED Start Stop Button 11 QI TAI M 11 OUTDO us eer sa ALD Hoang sa RUPEE VR addu 57 repeating pulses cccccccssssseseseeenees 22 sepalal E srie RT 11 SO CLIIIOS aoa a A ESE 60 SIngle THO G CP oxi andis 60 S PG esthetic p EN 11 58 triggering multiple files on different triggers Multi File mode 63 triggering multiple files on same trigger Batch mode 67 Troubleshooting 93 TTL autocreating Sync Out pulses 31 blanking signal 57 digital output eese 57 Sync Out settings ese 60 trigger settings esee 60 tFIOQBr ID E REED ERE da 56 triggering in Multi File mode 63 twisted pair cables 70 U U outputs aDOL usines be M uns A Pure 69 70 measuring with an oscilloscope 77 OSCITOSCODO i sri i sh ME dap us 70 volt
112. ted user friendly by entering the desired pulses rectangular ramp or sine waveforms into a worksheet Waveforms can be combined freely to create virtually any stimulus pattern Created stimuli are displayed in a WYSIWYG stimulus display All channels are set up separately For even more convenience repetitive stimulus patterns do not have to be entered separately but can be grouped and looped You can also import waveforms in ASCII file format and use them for stimulation You can print and save stimuli and comments for later use and documentation of your experiments Creating Stimuli Combine pulse types Stimulus protocol worksheet Use aseparate row You can repeat rows Menus Toolbar Generate and edit protocols for each pulse type and groups of rows F MC Sbhimulus I MC SEnwubus Main Menu Filestm Fie Edt STG Batch Signal View Window Settings Help Das ee amp Tim m STG s0002 Disconnect mi Programmed W Sweep IL 1 1 Z vonage Range 8000 mi Curent Range 1600 pA F MC Stimulus Main Plena NI stiri Last Download Cork Mode p Qutput Mode j Download Channel Download Sync Comment 80002 On F Voge ie 172 7 3 v 4 wih 2P3R4 ESI F3 Of C Current WowWee hee he oe Be i 8 Channel 1 er ee Se A A A A A A AA Puse eat tae e zm fien me J Tc NON UI pene rectangular 1000 Lv o cu ss n si mv B0 ps Td mv qus mv mv Nm HF TM mv f IE B pm 7 if i mv
113. tructure 1 File header 2 Data File header The first two non comment lines have to be the following lines Files for older versions of the import filter version 1 00 can be imported with the current version as well Multi Channel Systems MC Stimulus II ASCII import Version 1 10 The next three lines have to be the following lines in any order channels number output mode mode format format number Make sure you define the total number of channels for the used STG properly 4 channels for a STG with 4 channels 8 channels for a STG with 8 channels and so on according to the software version You can select the maximum number of channels during the installation of MC Stimulus On the Help menu click About MC Stimulus II to check your current software version 36 ASCII Import Export Example You have to include the following line in the ASCII file if you have installed MC Stimulus Il for STG2004 channels 4 Output mode is either voltage or current Example output mode voltage Six different format types are available 1 2 3 4 5 6 Example format 3 Data Preceding the data the output channel number has to be specified by the following line channel channel number You can specify analog output channels from 1 to 8 limited by the number of analog output channels of the connected STG of course The digital Sync Out channel number directly follows the maximum number
114. u can select the channels that you want to download or download the complete file You can start the stimulation directly after the download or start it manually with the software or hardware controls or start it on a trigger See also chapter Operating the STG for more information Warning Check the file thoroughly before downloading it on the STG The stimulus display is independent from the worksheet and from the download Verify that you download only the channels that you want to use Do not start the STG if you are not sure about the channel configuration or the nature of the downloaded file Select the channels for downloading under Download Channel and Download Sync Download Channel Iw 1 fw 2 fw 3 Iw 4 Iw 5 Iw 6 fw ME Download Sunc Iw 1 wo fw 3 fw 4 IV 5 i 6 iw ME On the STG menu click Download or click E on the toolbar to download only the selected channels onto the STG or Download and Start 9 to download only the selected channels and start the stimulation directly after the complete download Disable enable warning messages Cancel this option to disable the message boxes displaying compiler warnings during stimulus download The two error messages that can be disabled are Warning Signal length shorter than the maximum and Warning Non zero last amplitude value They may present a nuisance if you download multiple stimulus files in batch mode On the Settings menu select or deselect Enable
115. u like on the screen also outside the main window of the MC Stimulus Il program This is especially convenient when you are using two monitors 32 Programming Stimulus Protocols Select the option Auto Download if you want to adjust the stimulus intensity level on the fly Deselect the option if you want to download the changes manually later Select the option Change All if you want to adjust the stimulus intensity to the same level on all channels Deselect the option if you want to set individual channels to different levels Drag the slider with the mouse for adjusting the stimulus intensity in the range of 0 to 200 percent of the programmed stimulus amplitudes The WYSIWYG display is updated automatically The worksheet is not updated It always relates to the stimulus protocol that you programmed initially If Auto Download is selected changes are immediately downloaded onto the stimulator after releasing the slider and the stimulus protocol is restarted from the beginning with the adjusted stimulus intensity level If Auto Download was not selected you need to download the stimulus protocol manually for changes to take place You can click the Reset All button to set the level on all channels back to 100 percent Amplitude l E Chi I I I 0 Al 100 150 200 Lhe I I I I I I I I I I I I I 0 Al 100 150 00 Cha es I I I I I I I I I I I I I I 0 All 1
116. unctionality value time value time Format type 2 With this format you can repeat rows in the same way as described for the worksheet Group repeat is not available value time value time repeat Format type 3 This format type is available only in the current version of the import filter It allows you to use a third column of Value and Time value time value time value time repeat Example The following picture shows an ASCII stimulus file opened in a standard spreadsheet program B format 3 channel 1 value time value time value time repeat 200 20000 200 20000 T ZEE 2 40000 200 0000 200 20000 EDO 2 40000 D 17 channal 2 18 value time value time value time repeat 200 20000 200 20000 ZEE 2 40000 200 0000 200 20000 BID 2 40000 D 38 ASCII Import Export The same file would look like this in a standard text editor program You can see the tabs spaces and CR LF lz Demo file 2 3Multi Channel Systems MC Stimulus 1iA SCII import Version 1 10 2l 5rhannels 8 nmutput mode voltage gt 4 B format 3 3 1U channel 141 llvaTues time value time values times 127002700002 200 20000 O02 600002 24 i3 0s 400002 O02 Os Os Os Off 147002700002 200 20000 O02 6O000 24 1802 400002 O02 Os Os Os Off 16 S 17 channel 2 l value times value times values times 137002700002 200 2000
117. us or open a stimulus file before downloading In Multi File mode the corresponding file will be completely ignored There is no stimulus generator connected to the computer or the STG may be switched off Please check the computer connection and the status lamp of the STG There is no valid connection to an STG Download of data is not possible Please check the computer connection and the status lamp of the STG The last voltage current values of the named channels are not zero The current voltage levels are set to zero after the last value The last used values of these channels are kept until the STG is stopped 94 ASCII Import Troubleshooting These error messages relate to a wrong file format Please see chapter Supported File Format for more information MCStimulus MCStimulus E a E X AN Error wrong channel number at line 8 MCStimulus E x AN Error unexpected end of File na data header line Found at line 9 MCStimulus AN Error too Few data columns at line 12 MCStimulus EE xj AN value outside of Stimulus Generator range 8000 mv will be clipped The named line contains an unexpected character The channel number specification has to precede the data The channel number either lacks or the specified number does not exist for example number 13 Each data column must have a header value time or repeat The number of data columns has to
118. use the Batch Mode Important As long as you are in the Multi File mode the Standard trigger settings trigger inputs assigned to single channels will have no effect You also cannot close files if you are in Multi File mode Please leave the Multi File mode by selecting Standard under Mode in the Trigger Settings dialog box to use the standard trigger settings or before closing files Please read also chapter Extended Multi File Mode Trigger Settings File Mode Cont Made C Standard Multi File Extended Multifile On C Df Settings Trigger C Program Files Multi Channel Systeme C Stimulus ll Dema 1 stm Browse Trigger C Program Files hult Channel Systems ME Stimulus ID ema z stm Browse Triggers C Program Files Multi Channel Systema LO Stimulus IE emo 3 stm Browse Trigger C Program Files Multi Channel Systems ME Stimulus IAD emo 4 stm Browse Clear 1 Connect all necessary trigger inputs to TTL signal sources 2 Set up your stimulus protocols and save the files 3 Onthe Edit menu click Trigger Settings 4 Select the option Multi File 5 Under Trigger 1 click Browse and select the appropriate stimulus protocol that you want to assign to trigger 1 6 Repeat step 5 for all necessary trigger inputs Confirm the settings by clicking OK 7 All selected files are opened automatically 63 10 10 5 Stimulus Generator 4000 Series Manual Check all stimulus protocols before download Please
119. ut All other stimuli are applied once after their respective trigger event Trigger Settings File Mode Cont Mode Standard Multi File Extended Multifile C Un Off Settings g LR mum p C 25 DI E em C g C CI mm m C mn Td CI co e t P e c2 DE Ix n CT uw m wo uw co Trigger Trigger Triggers Triggerd Trigger Triggerb Trigger E E NE NE NE NE ME ee ME NE ME ME oe NE ME NE NE ME E NE NE ME E E NE NE NE ME ME oe NE NE ME NE E NE NE NE NE NE ME oe NE NE ME ME i E E NE NE oe NE NE ME ME E NE NE NE NE NE MEC oe NE NE ME ME E E NE ME NE ME ME oe NE NE ME NE i E NE NE NE NE ME ME oe NE ME ME NE Triggers Fa Cancel Repeating complete pulse protocols Gating Trigger Repeated stimulation of complete protocols can also be controlled by a gating trigger Channels associated with a Trigger input will be active as long as the condition of that Trigger is 1 HIGH and switched off as soon as the condition becomes 0 LOW Gating trigger function can be enabled in the Trigger Settings menu tigger event while sweeping on channel assigned to same trigger Stop Restart i C Trigger as Gate Single Trigger 24 Programming Stimulus Protocols 5 6 Autocreating Entries and Editing Columns Changing time units for a complete column 1 Click any Unit column he

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