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1. bg PR D D D D D D D D D D D D D D D D D D D D D e D D D D D D D D Se Sea Sie Ze Se This is useful for exporting the image for use in an external reporting tool or for other uses where a snapshot of the chart is needed Supported image formats are e Enhanced Metafile Format EMF e Windows Bitmap BMP RGB Encoded e Independent JPEG Group Format i fil TT JPEG JPG j i WE 50 e Portable Network Graphics Format PNG 16 million colors e Save Data View to File e Export Data File to Excel Worksheet Ec Export Data Pad Eo Excel Worksheet ks Export Data view to Clipboard E i CG Print S Sp oi Auto EUN e y j j Exit Auto backup a T Opens a new Export Data file to Excel worksheet i i window where auto See section 3 6 3 7 i estate Export Data pad to Excel worksheet Auto Backup See section 3 6 2 E j f 1 minute Export Data view to clipboard 6 2 minutes Same as Save Data view to file except that the image is placed in the clipboard and can be imported in any program using the paste command CTRL V E minutes 10 minutes hour CG C 30 minutes e No auto backup Y _ Print Data View P i x Print Comments Ue Print Data Pad Print gt Print Data view e Prints the Data view on the defauit printer Print gt Print comments Prints the comments on the default printer Print gt Print Data pad e Pr
2. Chapter 4 is a description of everyday handling of Tissue organ bath and the different kinds of mounting views available It also contains procedures describing general as well as daily maintenance of the Tissue organ bath unit e g weight calibration of the force transducers and cleaning instructions Chapter 5 is a list of equipment needed to setup a complete Tissue organ bath system The chapter also contains a list of accessories and spare parts available Appendices contain various additional information about Software installation Service Shipping instructions Buffer recipes System specifications and Fuse changing DMT Tissue organ bath system Model 50TOBS Contents Hade MAKS DII ITI UTENTE 3 Iddee d e DEE e SEN 5 EMC 7 7 EE 6 ADDFOVIIS E 6 Certificate Of COmfOrmity ccsccssecenscenseenseenseenseenseenseenseenssenseeeesenssenssenness 7 About this manual un ecceisnuss RUE sra Es ua Esa REV CHR cn uEE EUR CES EE uewsucteedeswededeedauencvuedse 8 Eih Tu dE 9 Unpacking the Tissue organ bath system esee 11 Chapter 1 System Overview o eeee eere rere nenne nnn nnn nnn 12 1 1 Tissue organ bath front panel eee re nen 12 1 2 Tissue organ bath rear panel eese eere eene nnn nnn 13 1 3 Tissue organ bath unit e eeeeeee eee ee eren nenne nnn nnn nnn 14
3. Asecond new method in the 7BOTOBS Is to T3 750TOBS Controller A perform 4 constant File Setup ew Wash About Register 7 T Connect to 750T0B5 oo r Tissue Chamber 2 r wash at a preset flow FSOTOBS paramaters Fill Empty calibration Bane 5 7 n rate In the 750TOBS Chamber 1 Presse TE the fresh warm buffer Chamber 2 Heating Plate temperature On from the reservoir Chamber 3 Temperature Difference a Zem al aa Chamber 4 Analog Output is filled from below ponn Force Units k Weight Calibration Data and excess buffer is User InterFace Wash evacuated from the Pie aliaki 1 Mode Foy Constagt Wash overflow on the top Calibrate Transducers 1 rends The flow rate can be programmed from 0 5 5 0 ml min Although it may take a little more time to completely wash any drugs away the advantages are that the tissue is never in air stays in warm buffer and stimulation if used does not need to be interrupted The user is encouraged to try both methods and based on own judgment determine what is best for the experimental conditions When the Constant wash mode is selected go to Setup Chamber 1 4 to adjust the settings T3 750TOBS Controller A File FE views Wash ae Re Connect to F50TOBS FSOTOBS paramaters A S 1 Charfiber 2 Chamber 3 d chamber 4 Force Units Chamber 1 T 5cale r Temp al Max value 50 Set Temp 37 0 Min valu
4. 0 13 40 00 13 45 00 13 50 00 Wee vi Ada y Chamber a Force 3 d Auto f Al a Zero a Tissue Chamber 2 Az Aulo a Zero HE Tissue Chamber 3 ASIE LEN a GE Tissue Chamber 4 EES IT Auto ei File C Documents and Settings Stine Dokumenter Organbad manual 07122006 2 Chamber 1 T 750TOBS Software File Setup View Wash About T QQ In QQ 4 44 4 Dd vm i amp W sse JI 7507085 A Comments Test tab Data Pad 13 30 00 13 35 00 13 40 00 13 45 00 13 50 00 n 12111 ze koe OP S el Add Tissue Chamber sis c Auto A a Zero me Tissue Chamber 2 KS Auto E Zero Nr Tissue Chamber 3 m Qs za d Auto a Zero Nr Tissue Chamber 4 gl Temp 53 Z A EN e Auto a Mr File C Documents and Settings Stine Dokumenter Organbad manuall07122006 2 Chamber 1 121112 2 D 750TOBS Software File Setup View Wash About YQ QQ Ma 4 vd vp amp OSM ol Ql Roe OP 8 TSUTOBS A Comments Test tab Data Pad 13 30 00 13 35 00 0 bey ttle j mouse D 13 40 00 13 45 00 13 50 00 n Tissue Chamber 1 ENSE Force 10 3 h Auto All a Zero PEE Tissue Chamber 2 z dis zl Auto a Zero HE Tissue Cham
5. All added comments can be seen in the comment sheet when clicking on the Comments tab By double clicking on a comment the view changes to the Dataview tab to the place where the comment was added The dashed line will flash for 5 seconds Experimental notes 13 750 TOBS Software File Setup View Wash About FQ amp pe G dd 4 rom pi Gy tee ala QI ole OBE 7E0TOBS A Comments Test tab Dats Pad n Sex Comments Sampling is started Sample Frequenzy i is set to 20 Hz O2 312 0 1 2 100 pM O2 330 M 0 1 M kel 2 gang Wash Emptying chamber Emptying charnber Emptying chamber Emptying chamber Stopped emptying chamber Stopped emptying chamber Stopped emptying chamber Stopped emptying chamber Filing chamber Tissue Chamber 1 JP ali m Auto A m EXE E Tissue Chamber 2 Ad Oa pe 8 Force 45 e ms I Auto a zen Fill r Tissue Chamber 3 y Gr Temp aT EN Force z E Auto a m Fill Tissue Chamber 4 ag Ta gens Bx Force Z 0 A GO n Ss D BR F P Auto E Zero Fill Fle C Documents and R a a E dat Chamber 1 4 1968928 Z Stopped filling chamber Filling chamber Connected to CO A comment can also be seen in the Dataview area Choose the select tool e and hold the cursor above a comment line Left click and a new window opens with data for the selecte
6. Chapter 2 Setting up e ceeieeeeeeie rere nee enne nnn nun nun KEEN 15 2 1 The Complete Tissue Organ bath system 750TOBS 15 2 2 Setting up step by step eeoeeeieeere nere nenne nnn nnne 15 Chapter 3 User software 1 11 eeeeeeeee rene nennen KEREN KENE EEN REENEN EN 16 3 1 The Software program 1 11 KENKRENERE eene nnne nn KEREN KENE EEN REENEN un 16 3 2 The 750TOBS Software User Interface eene nenne 16 3 2 1 Connect to EE 16 3 22 The EE TTe Kee 17 3 2 2 Drop down MENUS EE 20 CPP ES SM NIE INS ME 20 32 32 SCUD MENU SE 21 22 39 FII EMD UY Call bralo E 22 QE ANS E E 22 3 2 3 5 Heating plate feHTDel dll Ouiususesotusmisipiu cuia fac qa veteaanteuseategesectotaenstnieansesaresscnes 22 3 2 3 6 Temperature GiffEreNCe ccccesscnsescceescneeseneesecseeenensenseeesansenausanseeseaeesnaneseneess 23 3 2 3 AO ODD E 23 92 9 5 VG ISIE Calibration data E 23 Bi O E X DES 24 3 2 3 10 Flow CON e NER 24 dos Od T Mode E 25 De e I MEMINI TUNE 28 cnccu 7 eee ete U 4 0O0 0 EA E E A 28 S MU I E 29 3 5 1 The Dataview area uecaudexcuintazicnint duntuzuutuxztatanalucscuntudatacy ndc as tinc ueuc urscicha i nenne 29 Svo Be ak NONE UU TTE 29 3 CONIBIEDIES eege 30 E EREECHEN 31 3 6 1 Transfer data to the Datapad ssesccext asc
7. Double click Zero Z max 20 s sec Stop gt IES Q bk ocPtE SS Add r Tissue Chamber e v EN Temp Force 0 7 Auto Al M Zero Fil Tissue Chamber 2 AT Da Wa G Force 4 8 Aut a on Auto Zero Fill Tissue Chamber 3 Temp 27 35 i aT EN Force 3 5 Auto a Zero I Fi Tissue Chamber 4 AT EN Temp f2 7 6 Force 17 3 Auto a 08 30 00 08 32 00 Zero Connected to COM3 File C Documents and Settings Stine Dokumentertjvyhj dat Chamber 1 Mode WW Double clicking on the channel will also result in shifting between one or four channels Double clicking on the Fprce value will open the Weight calibration window 28 DMT Tissue organ bath system Model 750TOBS 3 4 1 Wash Washing e Number of washes left in the sequence Washes Left start washing d Stop washing D Se D D Ze Shortcut to the setup window 3 5 1 The Dataview area 3 5 2 Reading data Activate the Cursor button e A line and a box with the readings will appear on the datasheet Move the cursor by AR BIDE the left mouse button while moving the mouse along the curve e I 750T0BS Software u en File Setup View Wash About Za amp 4 44 4 gt p gt vi te Ex EAE ETT SES 750TOBS A Comments Test tab Data Pad H r Tissue Chamber 1 a y Temp ES EN Force 0 1 Auto Al A A Tissue Ch
8. Volume in a choosen chamber X axis color X axis setting Comments setup Up to 20 default comments can be saved The window can also appear by double clicking on the Comments input field User manual 750TOBS Chapter 3 21 3 2 3 3 Fill Empty calibration Fill Empty Calibration Setup gt 750TOBS parameters gt Fill Empty calibration Fill The amount of fluid filling the tis Chamber 1 300 mSec iml 350 mSec ml sue chamber is time regulated Chamber 2 300 m ml 350 m er jml Calibration of each chamber is performed with the marked tissue chamber provided with the calibra Chamber 4 28 m5ec ml 350 mec jml tion kit 20 ml Chamber 3 200 m er md 350 msec jml Adjust the numbers of each tissue chamber until the amount of fluid is correct the numbers indicates msec ml If the amount of fluid is to small the number msec ml must be higher If the amount of fluid is to high the number msec ml must be lower 3 2 3 4 Pressure Setup gt 750TOBS parameters gt Pressure Adjustment and on line reading of pressure This set Pressure Setup ting is normally not changed Set Pressure 0 400 Ke Actual pressure LJ Act Pressure provides an on line reading of the ac tual pressure in the reservoir bottles Set pressure To set or reset the target pressure 0 01 0 999 Bar Default 0 400 Update Update provides an on line reading of the actual pressure NOTE
9. 750TOBS Software A before 750TOBS Software B DMT Tissue organ bath system Model 50TOBS 3 2 2 The Main window e Data view tab T3 750TOBS Software A File Setup View Wash About GG e amp 4 4 b Pb Ed mac e Drop down menus Register Daten Comments Data Pad Connected to COMS e Dataview toolbar e Chamber controls Sex E Tissue Chamber 1 AT Fa e 364 A Fae 00 E Auto jf Al B Zero aj 7 SE Chamber 2 53 3 A Ta Temp EGERN AT K Force fo A ENT ik al ero r i 10 ER Se T F E Ei 5 Force 0 0 joo S T Chamber 4 ER em d zen IE Je I c d All data tootoo t Seee ee LC IET See eee fee T zs Ir e i ny SS p a EL T Chamber 1 4 e Statusbar Activate an axis by left clicking on it The activated axis is marked by a black rec tangle The axis can now be scrolled or zoomed using the scroll 4 or zoom amp tool Arrow keys PageUp PageDown Home and End buttons can also be used on an active axis Figure 3 1 The Tissue organ bath User Interface DataView toolbar 20 s sec v View all Increase X axis Decrease X axis View all Increase Y axis Decrease Y axis Go to the beginning of the file Go to the beginning one page at a time Go to the beginning 1 10 page at a time Go to the end of the fi
10. C 4 x BNC 2 5V FS filtered signal USB 2 0 110 240V 50 60 Hz User manual 750TOBS Appendix 3 45 Appendix 4 Fuse changing The main fuse of the Tissue organ bath system is placed inside the power inlet on the 7SOTOBS If the fuse blows it is easily changed using the following procedure Important When a fuse blows and needs to be changed it is imperative that the replacement fuse is equal to the one blown The 750TOBS system uses T2A 250V 6 3 x 32 mm DMT recommends that both fuses in the fuse block are changed at the same time as it can be difficult to determine which fuse is blown To replace the fuses Use a small screwdriver to open the voltage selector block Remove the red fuse block Remove the existing fuses Insert the new fuses Replace the fuse block back into the voltage selector block Note ensure that the correct voltage for your country is displayed pr geo Ios Voltage selector block 46 DMT Tissue organ bath system Model 750TOBS Appendix 5 Software installation Insert the 75OTOBS installation CD to install the 75OTOBS Control program Normally the sofware installation will start automatically If this does not happen run the program setup exe from the CD The following is a description of the installation procedure GBI 750TOBS InstallShield Wizard ES Welcome to the InstallShield Wizard for 7T50TOBS The InstallShield R Wizard will install 7SOTOBS on you
11. Click View gt Slide Up to hide the controller i i i i l File Setup View Wach About Register Click View gt Slide Down to show it again o Tissue Ch Slide L Connected to COMS 8 4 1 Wash Washing Washes Left f Start washing DMT Tissue organ bath system Model 50TOBS 8 4 2 About In the About box software firmware and hardware ID for the 750TOBS is shown There are also links to ASOTOBS Controller Acquisition Software Ver z 2 0 17 4 DMT support and website Firmware M DI De DP Firmware 1 01 06 000 Firmware 2 01 06 000 Firmware 3 01 06 000 Firmware 4 01 06 000 Hardware ID M 58256 Hardware ID 1 12000 Hardware ID 2 11963 Hardware ID 3 17044 Hardware ID 4 12062 Email zu orti dmt dk WW vene drat dk 8 4 3 Register It you want to register the software click Register and your Web browser will open It Tissue Chamber 3 gt o EE r Tissue Chamber 2 Em est Ee us EE T a ml Connected to COMS User manual 750TOBS Appendix 8 65 66 Appendix 1 Terms of warranty Warranty DMT A S warrants to the original user that myograph systems manufactured by DMT A S will be free from defects in materials and workmanship for a period of three years after the date of delivery DMT A S will repair or replace any defective part subject to the conditions limitations and exclusions Exclusions Force and pressure transducers s
12. IM A Bo e Oxygenation CO tubing pipe e Suction tube pipe Perfusion buffer inlet e Figure 1 3a Tissue organ bath unit NOTE Regarding the Oxygenation CO regulator The regulators need to be greased at least twice a year Also make sure that the regulators are turned at regular intervals to prevent them from getting stuck Use the grease that came with the system DMT Tissue organ bath system Model 50TOBS Chapter 2 Setting up 2 1 The Complete Tissue organ bath system 750TOBS CS200 Current stimulator optional Organ bath rear panel Connection to O CO supply Power supply Vacuum pump USB connection optional aa e Suction bottle optional Organ bath front panel PC data acquisition amp software Figure 2 1 Complete Tissue organ bath system 2 2 Setting up step by step 1 Oxygen CO supply The oxygen CO supply is attached to the pressure inlet on the back of the organ bath After connecting the main gas supply the regulators can adjust the inlet into each chamber Turning the regulator clockwise increases the oxygen CO supply turning it counter clockwise decreases it Pressure inlet Min 2 bar 200 kPa 29 PSI Max 8 bar 200 kPa 116 PSI 2 Suction connection The system has built in suction manifold with separate valves that allows each chamber to be drained individually After connecting your suction device to the outlet on the back of the Tissue organ bath al
13. Model 750TOBS ADI PowerLab data acquisition system Including Chart data acquisition and analysis software e Standard PC system DMT Item 80150 Constant flow shunt kit DMT Item 80150 CS200 4 channel Current stimulator DMT ltem CS 200 5 3 Tissue organ bath 750TOBS system spare arts This list contains a complete list of standard available spare parts for the Tissue organ bath 75OTOBS For parts not listed in this Chapter or for special parts which may need to be custom made please contact DMT for further information Force transducer e Calibration Kit e Tissue bath chamber size 5 ml DMT Item 75105 10 ml DMT Item 75110 20 ml DMT Item 75120 50 ml DMT Item 75150 e Mounting hooks rings Pins L shape 4 sets DMT ltem 75300 rings EFS Pins L shape w stimulation electrodes 4 sets DMT ltem 75310 bladder Closing clips 4 sets DMT ltem 75320 bladder EFS Closing clips 4 sets w stimulation electrodes DMT Item 75330 strips Triangle hooks standard 4 sets DMT Item 75340 strips EFS 4 sets Triangle hooks standard w stimulation electrodes 4 sets DMT Item 75350 Tubing kit DMT Item 75400 DMT Tissue organ bath system Model 50TOBS Appendix 1 Service Check A Tissue organ bath system working at optimal performance is extremely important for success To make sure that our customers always are dealing with first class Tissue organ bath systems DMT offers a Servi
14. Tissue organ bath system Model 50TOBS Unpacking the Tissue organ bath system Please take a few minutes to carefully inspect your new Tissue organ bath system for damage which may have occurred during handling and shipping If you Suspect any kind of damage please contact us immediately and we will take care of the problems as quickly as possible If the packing material appears damaged please retain it until a possible claim has been settled We recommend that you store the packing material for any possible future transport of the Tissue organ bath system In case of transport and the original packing material is unavailable please contact DMT Sales Department for advice and packing instructions After unpacking your new Tissue organ bath system please use the following list to check that the system is complete e 1x Tissue organ bath e 4x Chamber units 20 ml e 4x 1000mI Schott Duran Bottles incl special designed screw caps e 4x Tissue holders L 70 mm hooks e 1x Tube Kit e 1x Calibration Kit including marked tissue bath 20 ml and 5 gram weight e 1x USB cable e 1x Power cord e 5 meter cable air clamps e 2x mm fasteners Grease e 1x user manual e PC Acquisition software The shape of the AC plug varies by country be sure that the plug has the right Shape for your location User manual 750TOBS Unpacking the Tissue organ bath system 11 12 Chapter 1 System overview 1 1 Tissue organ bath front pa
15. and the colors of the dataview are changeable e The windows can be resized e Expanding or contracting the X and Y axis can be done to meet the needs of the user e Showing all four channels or just one channel There are three tabs 1 The dataview tab showing the data collected from the four force transducers 2 The comments tab showing a log of all user actions done on the four chambers 3 The Data Pad tab showing user chosen data from the dataview The acquired data comments and the Data Pad can be exported to an Excel Worksheet for further processing 3 2 The 750TOBS software User Interface The first time the DMT Tissue organ bath is turned on the VCP drivers must be installed as described in Appendix 6 3 2 1 Connect to 750TOBS Setup gt Connect to 750TOBS 13 No Connection LJ The connection to the 5OTOBS is automatic When the connection is performed the ho connection to E message Connected COMX is displayed in the status bar in the bottom left corner of the program window If the connection fails the message display is Not connected In case the connection fails try connecting by clicking on Setup gt Connect If the connection fails again check if the USB cable is mounted correctly and 750TOBS is turned on Check that the USE cable is connected and the power is turned on The following COM ports are available If two 75OTOBS are connected to one PC use both programs Always connect
16. contains goods returned for repair service Customers outside the EC must further enclose a proforma invoice stating that the box contains goods being returned for repair or service If arranging transportation through a courier please keep in mind the high value of the Tissue Organ Bath System and that a standard insurance provided by the courier in most cases is insufficient to cover damage or loss of the system In most cases an additional insurance coverage is needed DMT Tissue Organ Bath System Model 750TOBS Appendix 3 System Specifications Specifications Chamber Chamber size s Chamber material Chamber heating Temp range Temp resolution Temp stability Chamber suction Chamber aeration Chamber filling Chamber filling method Transducer Force range Output resolution Transducer calibration Reservoir Reservoir volume Reservoir heating Inlet pressure Safety valve Temp range Temp resolution Temp stability Analogue output s Data communication Voltage 5 10 20 or 50 ml Glass Heated air circulation built in patent pending lt 50 C 0 1 C 0 1 C Manual or automatic via software Individually via precision needle valve s Manual automatic via software Drain fill or constant wash 200 mN 0 1 mN using filtered signal Semi automatic via software 4 x 800 ml Electronically built in Min 2 0 bar max 8 0 bar 10 bar lt 50 C 0 1 C 0 2
17. installed Please wait while the InstallShield Wizard installs 7S0TOBS This may take several minutes Status Installshield The 750TOBS Control program is now installing from the CD The InstallShield Wizard has successfully installed 750TOBS Click Finish to exit the wizard Click Finish The control program is now ready to use Start the program from the icon on the Desktop or via Start gt All Programs gt DMT gt 750TOBS folder lt Back DMT Tissue organ bath system Model 50TOBS Appendix 6 VCP Driver installation The first time the 75OTOBS is connected to the PC the VCP drivers must be installed They are on the CD delivered with 5O0TOBS To install VCP drivers for 5bOTOBS under Windows XP follow the instructions below When the USB cable is plugged into the back of 5OTOBS for the first time this window will open Choose No not this time and click Next Select Install from a list or specific location Advanced as shown below and then click Next Found New Hardware Wizard Welcome to the Found New Hardware Wizard Windows will search for current and updated software by looking on your computer on the hardware installation CD or on the Windows Update Web site with your permission Read our privacy policy Can Windows connect to Windows Update to search for software Yes this time only Yes now and every time connect a device No not th
18. sales dmt usa com Support dmt usa com
19. to reach pH 7 4 User manual 750TOBS Appendix 7 53 54 7 1 3 KPSS 1 x KPSS 1000ml Solution 1 Chemical MW g mol Conc mmol l Conc g l KCl 14 56 123 70 9 223 MgSO H O 246 48 1 17 0 289 KH PO 136 09 1 18 0 161 Solution 2 Chemical MW g mol Conc mmol l Conc g l CaCl 2H O 147 02 2 50 0 368 Solution 3 Chemical MW g mol Conc mmol l Conc g l NaHCO 84 01 25 00 2 100 EDTA 372 24 0 03 0 010 Glucose 198 77 5 50 1 091 1 Dissolve the chemicals in approximately 100 ml double distilled H O as three individual solutions as described in the table above Gently heat solution 3 to dissolve the EDTA Solution 1 is added to a graduated bottle and the bottle is filled with double distilled HO to a final volume of 500 ml Solution 3 is added to the graduated bottle that is filled with additional double distilled HO to a final volume of about 850 ml Aerate the solution with carbogen 9576 O 5 CO for about 20 minutes Solution 2 is added and the graduated bottle is filled with additional double distilled H O to reach the final volume of 1000 0 ml Continue the carbogen bubbling until the pH of the buffer solution reaches 7 4 DMT Tissue organ bath system Model 50TOBS 25 x KPSS 1000ml Solution 1 Chemical MW g mol Conc mmol l Conc g l KCI 14 56 123 70 230 575 CaCl 2H 0 147 02 2 50 9 200 Solution 2 Chemical MW g mol Conc mmol l Conc g l MgSO H O 246 48 1 1
20. to setup an Organ bath system In addition the chapter contains a list of special Tissue organ bath 5OTOBS accessories and spare parts Besides the main focus on development and manufacturing DMT has specialised in offering our customers first class laboratory equipment needed for a lab operating a tissue bath set up at very competitive prices Please contact DMT Sales Department for further product information and prices 5 1 General Tissue organ bath equipment This list contains a complete checklist of laboratory equipment needed when setting up a Tissue organ bath system Dissection stereo microscope DMT recommends the Zeiss Stemi 2000 stereo microscope Dissection scissors DMT Item DS 1000 DMT recommends the Geuder G 19745 8 cm straight scissors e Dissection and mounting forceps DMT Item DF 3000 DMT recommends the Dumont medical No 5 and 55 tip 0 10 mm x 0 06mm Pipettes DMT recommends CappAero pipettes e Schott Duran glass bottle 1000ml other bottles may not fit e Vacuum pump DMT recommends a membrane vacuum pump having a suction volume of at least 11 litres per minute Dissection petri dish DMT Item PD 2000 DMT recommends a 9 cm glass Petri dish coated with a 5 mm Sylgard polymer layer e Grease DMT ltem HVG 1000 User manual 750TOBS Chapter 5 41 42 5 2 Tissue organ bath system accessories This list contains a list of special accessories available for the Tissue organ bath system
21. 030 USA China Tel 45 87411100 Tel 86 0 21 64869685 Tel 1 770 612 8014 Fax 45 87 411101 Fax 86 0 21 64280591 Fax 1 678 302 7013 www dmt dk www dmt asia com www dmt usa com sales dmt dk sales dmt asia com sales dmt usa com support dmt dk supportedmt asia com supportedmt usa com User manual 750TOBS Trademarks Introduction The vertical organ bath with its double walled glass chamber is the traditional ex perimental setup that has been used extensively to investigate the physiology and pharmacology of smooth muscle vascular and non vascular and other tissue prepa rations through measurement of contractile force while bathed in an appropriate physiological solution or buffer at 37 C The advantages in these studies is that the preparation can be subjected to pharmacological agents while excluding the influence of systemic processes that occur in intact animals The results of these organ bath systems are generally more consistent reproducible and lend themselves for screen ing and the measurement of concentration response curves CRCs The most common application of the organ bath assay are in cardiovascular research using isolated aorta rings heart tissue papillary muscle or arteries In studying gastro intestinal effects preparations of ileum and colon are often used but also gastric antral muscle and sphincter may be studied Respiratory effects can be studied in isolated tracheal rings phrenic diaphragm preparat
22. 2 3 12 Weight calibration By activating Setup gt 750TOBS parameters gt Calibrate transducers gt Chamber X a message appears as shown to the right Place the 5g weight on the transducer Wait until the relative force reading is stable Click OK The force applied on the force transducer should mimic the stretch created by the contraction of a mounted ring preparation 3 3 1 View Weight Calibration Activate Force 1 with 5 gram Then push OF to calibrate Cancel The Dataview can be set to display all four channels or one single channel D 750 TOBS Software anne 1 4 ecol e channel 2 e Channel 3e Channel KE TAA 750TOBS Comments De B TQ Qi kel amp 4 44 4 P fN Sai so lp Ql Roe OP EB S0TOBS A Comments Test tab Data Pad 0 m Tissue Chamber 1 atii E T R II4R XeuI 2 44 wo All B D T50TOBS Software File Setup View Wash About 3 Q amp Gd 44 4 b DP A 750TOBS A Comments Test tab Data Pad 0 08 24 00 08 26 00 08 28 00 Connected to COM3 File C Documents and Settings s 08 28 00 z Add eS al Temp ES Force 0 7 T3 750TOBS Software File Setup View Wash About EF QA handie aa 4 b Pb Pi a 208 Channel 1 7SOTOBS el gt tab Data Pad Channel 3 Channel 4 Enable comments to see where comments have been added See Paragraph 3 2 3 2
23. 39001 40048 0 5 0 1 0 9 0 1 19 39001 40048 0 5 0 1 0 9 0 1 20 39001 40048 0 5 0 1 0 9 0 1 21 39001 40048 0 5 0 1 0 9 0 1 22 39001 40048 0 5 0 1 0 9 0 1 23 39001 40048 0 5 0 1 0 9 0 1 24 39001 40048 0 5 0 1 0 9 0 1 25 39001 40048 0 5 0 1 0 9 0 1 26 39001 40048 0 5 0 1 0 9 0 1 27 39001 40048 0 5 0 1 0 9 0 1 28 39001 40048 0 5 0 0 9 0 1 29 39001 40046 0 5 0 1 0 9 0 1 The data in column A is in a data time format used in Microsoft Excel It can be formatted in the following way Mark column A right click choose Format Cells choose the Number tab choose Time Now it is possible to chose time format Note To show the data time correct in column A it is important that the setting under Control panel gt Regional options gt General is set to English DMT Tissue organ bath system Model 50TOBS 3 4 2 About In the About box software firmware and hardware ID for the 50TOBS is shown There are also links to DMT support and website YSOTOBS contraller amp Acquisition Software Ver Z z 0 17 Firmware M 01 06 0600 Firmware 1 01 06 000 Firmware 2 01 06 000 Firmware 3 01 06 000 Firmware 4 01 06 000 Hardware ID M 58256 Hardware ID 1 12000 Hardware ID 2 11969 Hardware ID 3 12044 Hardware ID 4 12062 dp MT NOSE WW ver drat dk 3 4 3 Register If you want to register the software click Register and your Web browser will open It is now possible to register at DMT s
24. 7 12225 KH PO 136 09 1 18 4 025 Solution 3 Chemical MW g mol Conc mmol l Conc g l EDTA 3 2 24 0 03 0 250 1 Dissolve the chemicals for solution 1 in about 800ml double distilled HO ina 1000 ml graduated Dissolve the chemicals for solutions 2 and 2 3 in 75 ml double distilled H O in individually cylinders Gently heat solution 3 to dissolve the EDTA 3 Solutions 2 and 3 are added to solution 1 and the graduated bottle is filled with additional double distilled H O to reach a final volume of 1000 ml Before use 4 Dilute the 25 x PSS stock solution 1 25 using double distilled H O 5 Add 1 091 g l Glucose 2 100 g l NaHCO 6 aerate the solution with carbogen 9576 O 576 CO for at least 20 minutes If necessary wait for the pH of the buffer to reach pH 7 4 User manual 750TOBS Appendix 7 55 56 Appendix 8 750TOBS controller Appendix 8 is the 5bOTOBS Software manual without the Data acquisition This software is used if the user prefers a different Data acquisition program 8 1 The 750TOBS Software User Interface The first time the DMT Tissue Organ Bath is turned on the VCP drivers must be installed as described in Appendix 6 8 1 1 Connect to 750TOBS Setup gt Connect to 750TOBS The connection to the 750TOBS is automatic LY Mo Connection Max When the connection is performed the message Connected COMx is displayed in the status bar in the bottom left corner of the program windo
25. Click the OK button Flow calibration for the chosen chamber is now finished Repeat step 1 5 for all the chambers 3 2 3 11 Mode select Flow Calibration 33 3 sec Setup gt 750TOBS parameters gt mode gt The 750TOBS has two different wash modes Constant wash and Fill empty The classic way of changing solution in an organ bath system is to drain the entire bath and refilling it with fresh buffer via a Fill empty routine The advantage of this method is that the concentration of drug declining fast especially if this is repeated 3 times or 13 750TOBS Software A File Setup View Wash About Connect to 750TOBS lag 4 b bb B E D Start FSOTOBS paramaters d Fill Empty calibration Chamber 1 Chamber 2 Chamber 3 Chamber 4 Axis amp Grid Comments Force Units Pressure Heating Plate temperature Temperature Difference 4nalog Output Weight Calibration Data Wach Flow Calibration i Mode k Constant Wash bow Fill Empty Calibrate Transducers more The 75OTOBS can do this automatically via the wash menu and the buffer comes from the pre heated buffer reservoir in the system Therefore compared to the classic approach washing is easy convenient and fast A disadvantage of this fill empty method is that the tissue is briefly suspended in air without nutrients and stimulation when used needs to be suspended When the fill empty mode is selected go to Setup gt Chamber 1 4 or double click on th
26. OM Include this location in the search C VCP Drivers v Browse Dont search will choose the driver to install Choose this option to select the device driver from a list Windows does not guarantee that the driver you choose will be the best match for your hardware Hardware Installation The software vou are installing for this hardware USB High Speed Serial Converter has nat passed Windows Logo testing to verfu its compatibility with Windows ab Tell me why this testing is important Continuing your installation of this software may impair or destabilize the correct operation of your system either immediately or in the future Microsoft strongly recommends that you stop this installation now and contact the hardware vendor for software that has passed Windows Logo testing Continue Anyway Found New Hardware Wizard Please wait while the wizard installs the software e USB Serial Converter ftdiunin exe To CNWwINDU WwS system32 50 DMT Tissue organ bath system Model 750TOBS Windows should then display a message indicating that the installation was successful Click Finish to complete the installation This has installed the serial converter The COM port emulation driver must be installed after this has completed After clicking Finish the Found New Hardware Wizard will continue by installing the COM port emulation driver The procedure is the same as that above fo
27. S Chapter 3 33 3 6 3 Export a Data file to an Excel Worksheet All the Data from the Dataview can be exported to a 50 file format that can be opened in an Excel Worksheet T 750TOBS Software A File Setup view Wash About New Open Save Click here to open the Save Data View to File PR Export settings dialogue Export Data File ta Excel Worksheet 4 Setup Comer nen TO ee ea l e Save Export Data View to Clipboard B PM l In the dialogue box Print the resolution of the Auto Backup data is defined In this Exit example every 5th datapoint is saved as File gt Export Data File to Excel Worksheet gt Save opens jan average value of 5 a standard Save file dialogue window Give the file a datapoints name and click save Export settings Export every 9 Open the 750 file in a Microsoft Excel worksheet y MICTOSOTT Excel C 2U File Edit View Insert Format Tools Dats Window Help AE Ere E Bi a M Arial 10 B Z U o A A B2 s XV k A B C S E E G H J K L M Experiment Test 3 Age Sex Comment 1 Comment 2 Comment 3 Comment 4 Comment 5 Comment 5 10 Comment 7 11 Comment 8 12 Comment 9 13 Comment 10 14 Time Chamber 1 Chamber 2 G amber3 Chamber 4 Comment loo CO ooo Na 15 AJR 39001 40047 0 0 0 0 All Sampling is started Sample Frequenzy is set to 20 Hz 17 39001 40047 0 5 0 1 0 9 0 1 18
28. Tissue Organ Bath System Model 750TOBS with Data acquisition software User Manual Version 2 2 DANISA M170 T CHANOLOGS S7 DANISA M170 T e cc rETr COL COCS DMT Tissue organ bath system Model 50TOBS Tissue Organ Bath System Model 50TOBS User manual Trademarks PowerLab and LabChart are registered trademarks of ADInstruments Pty Ltd The names of specific recording units such as PowerLab 4 25 are trademarks of ADInstruments Pty Ltd Pentium is a registered trademark of the Intel Corporation Windows Windows 95 Windows 98 Windows ME Windows NT Windows 2000 and Windows XP are registered trademarks of Microsoft Corporation All other trademarks are the properties of their respective owners DMT reserves the right to alter specifications as required This document was as far as possible accurate at the time of printing Changes may have been made to the software and hardware it describes since then New information may be supplied separately This documentation is provided with the DMT Tissue organ bath system Model 750TOBS Version 2 2 No part of this document may be reproduced by any means without the prior written permission of DMT A S Copyright 2008 DMT A S DANISA MYO TECHNOLOGY DMT A S DMT Asia DMT USA Inc Skejbyparken 152 Everwin Gardens 1201 Peachtree Street DK 8200 Aarhus N Rm 502 Block B 400 Colony Square Suite 200 630 Denmark 521 Wanping Nan Lu Atlanta GA 30361 Shanghai 200
29. When the pressure is reset press the small button that appears to the right of the Set Pressure Input field If the pressure is changed the flow must be calibrated again as described in paragraph 3 2 3 10 3 2 3 5 Heating plate temperature Setup gt 750TOBS parameters gt Heating plate temperature Adjusts the temperature on the heating plate on which the reservoir bottles are placed The temperature here is chosen to be higher than the bath temperature to avoid gas formation in the lines and to correct for cooling during refilling of the chambers Set Temperature is used to set the set temperature of the heating plate for the reservoir bottles Default Heating Plate Temperature 39 C um The Set Temp is updated when clicking the OK button SUGES B9 0 3e NOTE To minimize the heating time one can benefit by pre heating the buffer to 35 C before filling the reservoir bottles for the first time in the day DMT Tissue organ bath system Model 50TOBS 3 2 3 6 Temperature difference Setup gt 750TOBS parameters gt Temperature difference Adjustment between the actual temperature and target temperature Chamber 1 2 9 Chamber 2 0 6 The Set temperature and Actual temperature may not be equal due to the ambient temperature in the lab It is possible to compensate this with this setting Example If the Set Temp for Chamber 1 is 37 0 C and the actual temperature measured in Chamber 1 is 36 2 C The Diff
30. amber 2 a y y v Temp ES E Force UR Auto a Zeb md n ill Tissue Chamber 3 a E Force 36 Auto a Zero A Tissue Chamber 4 Al 9 E Temp bo M Force jj a of a oof af Qe ft Tq Ins ee 09 37 00 09 38 00 09 39 00 09 40 00 09 41 00 09 42 00 09 43 00 09 44 00 On Bo Zao Fil Connected to COM3 File C ADocuments and SettingslStinelDokumenterlOrganbadYmanuallcarsten dat Chamber 1 968928 Z The automatic data scrolling on the X axis is stopped when the Select Button W or the Cursor button has been activated The automatic scrolling is started again by activating the Resume button gt 12 750TOBS Software 13 750TOBS Software File Setup View Wash About File Setup View Wash About i3 QQ amp id 44 4 eg MS TAA G GQ 4 44 4 P Pb A POOTOBS 4 Comments Test tab Data Pad ee i ee Pens xs Channel I Channel d lt All gt Style My value x v Style hey chil Optionbg Value X Options Ch z Snap to Data Poink Value Y Snap to Data Point Ch 3 Edit Period Edit Ch 4 Peak Peak Frequency Change the channel on which the Change the style on which the data data is read by right clicking on the is read by right clicking on the cursor cursor line line User manual 750TOBS Chapter 3 29 30 3 5 3 Comments tab Adding comments to the Dataview During the data acquisition comments can be added eg when the
31. ber 3 Ae oN Force 3 9 Auto a Zero Ge Tissue Chamber 4 ER ag 9 Temp 53 E E EN Force jes Auto W as Ge File C Documents and SettingslStinelDokumenterlOrganbadlmanuall07122006 2 Chamber 1 121112 4 DMT Tissue organ bath system Model 50TOBS Tissue chambers controls e Fill chamber Empty chamber r Tissue aar E h La E eg ELE checked the chamber is filled up with the programmed i N Zero force e amount of ml 4 Green The chamber Otherwise the 7 isin place chamber is filled Red The chamber until the fill up H is missing or not button is clicked S positioned correctly again If All i checked the activation of Fill Empty Zero Pressure and Fill of chamber 1 will be applied to all the chambers User manual 750TOBS e Pressure is activated deactivated by clicking the offe button below or Red The pressure is Soff Blue The pressure is on When the reservoir bottles are refilled or filled for the first time write the volume in the bottle in the window that pops up Chamber 1 Fill Volume 797 ml Chapter 3 19 3 2 3 Drop down menus 3 2 3 1 File menu New e Create a new file to begin a Data acquisition Open e Open an exciting file sg D D D D Se T3 750TOBs Software A E File setup View Wash About
32. ce check at a very favourable price The Tissue organ bath system Service check includes a complete separation for inspection of all mechanical and electronic parts The Tissue organ bath is then reassembled adjusted and finally all electronic and mechanical parts are tested Please note that the service does not include replacement of transducers or any other needed spare parts Please contact DMT for information about prices User manual 750TOBS Appendix 1 43 44 Appendix 2 Shipping instructions If the Tissue organ bath system needs to be returned for service or repair please read the following shipping instructions very carefully Before you start packing the Tissue organ bath system please remember that you are dealing with very delicate equipment and therefore care must be taken DMT recommends that each part of the system be wrapped individually i e with bubble wrap and placed together in a large box preferably the box you once received the tissue bath system in Place the wrapped items in the middle of the box and fill out the surroundings with chips of expanded polystyrene Important Before closing the box make sure that no enclosed items can be shaken around as transport by road or air from time to time is quite rough Address the box to DMT A S Skejbyparken 152 DK 8200 Aarhus N Denmark Make sure that all four sides of the box are marked fragile or similar Make an indication on the top of the box that it
33. change to the Dataview tab with the selected values centred in the window Licks Ele Setup View Wash abut IF amp amp b amp amp Data View Comments Data Pad Id dd 4 b PP b a DP e do Dese a sisi 1 x d 0 Tissue Chamber 1 ES 01 25 09 54 22 01 25 09 54 24 F 01 25 09 55 31 01 25 09 55 49 01 25 09 55 51 01 25 09 55 59 01 25 09 54 25 01 25 09 54 27 01 25 09 54 21 01 25 09 54 24 01 25 09 54 12 01 25 09 54 15 01 25 09 54 12 01 25 09 54 14 01 25 09 54 10 01 25 09 54 11 14 1182 16 9215 18 5117 16 9419 14 1182 Temp i 56 D Force Auto All m Zen Fi The chosen data from the Data view is transferred to the Data Pad as Comment Chamber Min Max Mean STD SEM Samples Start time End time DMT The comment is the nearest comment to the left of the transferred data The chamber where the data is captured from The minimum value of the transferred data The maximum value of the transferred data The mean value of the transferred data The Standard Deviation of the transferred data The Standard Error of Means of the transferred data The number of samples in the transferred data The start time of the transferred data The end time of the transferred data Tissue organ bath system Model 50TOBS 3 6 2 Export a Datapad file to an Excel worksheet All the Data from the DataPad can be exported to a 50 file format choose text txt format in excel t
34. ct for cooling during refilling of the chambers Set Temperature is used to set the set temperature of the heating plate for the reservoir bottles Default 39 C m The Set Temp is updated when clicking the OK button ses lens 59 0 SE NOTE To minimize the heating time one can benefit S SE by pre heating the buffer to 35 C before filling the reservoir bottles for the first time in the day DMT Tissue organ bath system Model 50TOBS Heating Plate Temperature E 8 2 2 5 Temperature difference Setup gt 750TOBS parameters gt Temperature difference Adjustment between the actual temperature and target temperature Chamber 1 2 9 Chamber 2 0 6 The Set temperature and Actual temperature may not be equal due to the ambient temperature in the lab It is possible to compensate this with this setting Example If the Set Temp for Chamber 1 is 37 0 C and the actual temperature measured in Chamber 1 is 36 2 C The Difference setting for Chamber 1 must be O 8 Chamber 3 0 8 Chamber 4 0 9 8 2 2 6 Analog output Setup gt 750TOBS parameters gt Analog output Analog Output The output is from O O to 2 5 Volt Resolution is 0 61 mV The span in this window can be adjusted LO Vgl 10 mM In the example above the range is set to 110 mN The output is O O Volt if the force is TOmN and 2 5 Volt if the force is 2 5 Volk LOO mM 100mN Direct Output The internal sample frequency is 20H
35. d comment EP b La 20 s sec Start d Ky pla EF Chamber E Time 38 06 Comment Stopped emptying chamber 2 36 00 09 37 00 09 38 00 03 39 00 09 40 00 3 41 User manual 750TOBS Chapter 3 31 3 6 1 Transfer Data to the Datapad A specific data area in the channel can easily be transferred to datapad Select the data to be transferred by holding the CTRL key while dragging the mouse over the area where the data needs to be collected When the left mouse button is released a Transfer to datapad window appears Click Yes The data can now be found in the DataPad tab Connected to COM3 D 750TOBS Software A D 750TOBS Software File Setup View Wash About QQ amp 44 4 gt gt amp UBER jse 2 Q kok DP 750TOBS Comments Test tab Data Pad D EH 77 Tissue PN ez ead cocco EU PRESSER TT EE See KE ll See pn S ere e K Force jos l CTRL FI VDS use button drag T Auto As EE GENEE i Ne TT EE I EET 172 i O eE S E er eee ere I Em iH dT T AA deb Ce 8 sei Gi m 20 0 MM Wi PE S Le ag Mil Sr m NS LS m ZC ZZ 09 03 00 09 04 00 09 05 00 osgo 09 07 00 09 08 00 09 File C Documents and g ttings Stine Dokumenter Organbad manual jgjh dat Chamber 3 Double Click on the line and the view will
36. e 0 Flow 05 mine OAM IA User InkberfF LO o OF Cancel DMT Tissue organ bath system Model 50TOBS Important When the mode is changed the shunt inside the bottle must be changed too The Constant wash mode requires the Constant flow shunt and Fill Empty mode requires the Fill Empty flow shunt Important Make sure that the pressure is OFF before changing the shunts A warning noting that the pressure is turned off is Attention Fr shown when the mode is changed A You are changing mode Please check that the right shunt is mounted in Ehe bottle Please note that the pressure is removed Constant flow shunt Hypodermic needle 0 4 x 40mm ee Fill Empty flow shunt A Accessories please contact DMT for purchase 8 2 2 11 Weight calibration By activating Setup gt 750TOBS parameters gt Calibrate transducers gt Chamber X a message appears as shown to the right Activate Force 1 with 5 gram Place the 5g weight on the transducer Wait until Ther push Ok te calibrate the relative force reading is stable Click OK eni The force applied on the force transducer should mimic the stretch created by the contraction of a LM mounted ring preparation Weight Calibration User manual 750TOBS Appendix 8 63 8 3 1 View Hide show the controller T3 750TOBS Controller A
37. e checked the chamber is filled Red The pressure is Soff Blue The pressure is on Zen up with the 1 i rogramm i E e Geh SC i G The ch i EES When the reservoir Green The chamber Otherwise the Jj sci DECH bottles are refilled chamber is filled i or filled for the first until the fill up d a rae time write the volume D is missing or no button Is clicked S positioned correctly a eee again j window that pops up d If All is checked the activation Fill Volume TN m of Fill Empty Zero Pressure and Fill of chamber 1 will be applied to all the chambers 8 2 2 Drop down menus 8 2 2 1 Setup menu Connect to 750TOBS e ee Sa T3 T50TOBS Controller A e Force units Choose the 750TOBS T He Setup View Wash About between mN or gram E d e Connect to 750TORS L i 750TOBS parameters e ie DUT paramaters e User Interface There See sections 8 2 2 2 to C Chamberi i are two choices 69944 psu if 1 Only Controller ponds M 2 Acquisition amp Corp aud F Controller Described Force Units ke i in Chapter 3 User InterFace ka User manual 750TOBS Appendix 8 57 8 2 2 2 Fill Empty calibration Fill Empty Calibration Setup gt 750TOBS parameters gt Fill Empty calibration Fill The amount of fluid filling the tis Chamber 300 mSec ml 350 mec rl sue chamber is time regulated Chamber 2 300 mote jml 350 m er jm
38. e which afterwards is filled with additional double distilled H O to a final volume of 850 ml 4 Aerate the solution with carbogen 9576 O 576 CO for about 20 minutes 5 Solution 2 is added and the graduated bottle is filled with additional double distilled H O to reach the final volume of 1000 ml Continue the carbogen bubbling until the pH of the buffer solution reaches 7 4 52 DMT Tissue organ bath system Model 75OTOBS 25x Concentrated PSS 1000ml Solution 1 Chemical MW g mol Conc mmol l Conc g l NaCl 58 44 118 99 173 850 KCI 14 56 4 69 8 750 CaCl 2H 0 147 02 2 50 9 200 Solution 2 Chemical MW g mol Conc mmol l Conc g l MgSO HO 246 48 1 17 5225 KH2PO 136 09 1 18 4 025 Solution 3 Chemical MW g mol Conc mmol l Conc g l EDTA 372 24 0 03 0 250 1 Dissolve the chemicals for solution 1 in about 800 ml double distilled H O in a 1000 ml graduated bottle Dissolve the chemicals for solutions 2 and 3 in 75 ml double distilled H O in individually cylinders Gently heat solution 3 to dissolve the EDTA Solution 2 and 3 is added to solution 1 and the graduated bottle is filled with additional double distilled H O to reach a final volume of 1000 0 ml Before use Dilute the 25 x PSS stock solution 1 25 using double distilled H O Add 1 091 g l Glucose 2 100 g l NaHCO Aerate the solution with carbogen 95760 5 C0 for at least 20 minutes If necessary wait further for the pH of the buffer
39. e chamber s y axis to adjust the settings 13 750 TOBS Software A File Setup View Wash About C Connectto750TOBS qq d P pp b n FSOTOBS paramaters k e Chamber 1 ad R Chamber 2 Chamber 3 Chamber 4 Axis amp Grid Comments m Dee 2 Q s Force Units k Chamber 1 I Y Scale gt Temp ol Mas value 50 Set Temp 37 0 Min value O Volume 20 m SUE retener entente Vase A AI EN al OK User manual 750TOBS Chapter 3 25 26 A second new method in the 75OTOBS is to perform a constant 13 750 TOBS Software A File Setup View Wash About Connectto750TOBS dd d b bb b 20s si Chart wash at a preset flow 350TOBS paramaters gar ETRE k rate In the 50TOBS Chamber 1 Pressure the fresh warm buffer Chamber 2 Heating Plate temperature from the reservoir Chamber 3 Temperature DiFFerence ped Chamber 4 Analog Output is filled from below Axis amp Grid Weight Calibration Data and excess buffer is Comments Wash evacuated from the Force Units Flow Calibration CH rfi nthet 1 EE k w Constant Wash PANEM SEN Calibrate Transducers FillEmpty The flow rate can be 2 m z programmed from 0 5 5 0 ml min Although it may take a little more time to completely wash any drugs away the advantages are that the tissue is never in air stays in warm buffer and stimulation if used does not need to be interrupted The user is encouraged to try both
40. e tubing inside the reservoir bottle Calibration of flow in Constant Wash mode is performed by the marked Tissue chamber provided with the calibration kit 5 ml On the Tissue chamber there are two marks used for the calibration The calibration is done one chamber at the time 1 Turn off oxygenation 1 EN ud 2 Choose the chamber to calibrate SE Click Start flow SE Mises Osec Cancel 3 When the surface of the fluid reaches the bottom mark on the calibration tissue chamber click the Start 7 EY 1 Measure button Check this by looking Geelen 2 at the line at eye level and look for the Start Measure i Start Measure O sec bottom of the fluid meniscus to reach Se the line m Ok Cancel Flow Calibration the top mark on the calibration tissue chamber click the Stop Measure button Flow Calibration 4 When the surface of the fluid reaches top Measure DMT Tissue organ bath system Model 50TOBS 5 Click the OK button Flow calibration for the chosen a Flow Calibration chamber is now finished Hee E nS Stop Flow Repeat step 1 4 for all the chambers 33 3 56C 8 2 2 10 Mode select Setup gt 750TOBS parameters gt mode gt The 750TOBS has two different wash modes Constant wash and Fill empty The classic way of changing solution in an organ bath system is to drain the entire 13 750TOBS Controller A File Setup View Wash About Registe
41. eparately or part of myograph systems manufac tured by DMT A S are disclaimed from any warranty Limitations This warranty shall not apply to equipment subjected to accidental damage improper use alteration or deterioration Warranty on third party products will be as determined by their respective manufac turer DMT A S shall not be liable for consequential incidental special or other direct or indirect damages resulting from economic loss or property damage sustained by you or any end user from the use of the products sold or services rendered hereunder Warranty returns A Return Material Authorisation RMA number is required for all returns This number should be clearly indicated on all returned myograph systems Products damaged due to improper or inadequate packaging when returned for RMA purposes are not granted warranty coverage DMT Tissue organ bath system Model 50TOBS DANISA M COT CEFN COOLU_ OGS DMT A S Skejbyparken 152 DK 8200 Aarhus N Denmark Tel 45 87 41 11 00 Fax 45 87 41 11 01 www dmt dk sales dmt dk support dmt dk DMT Asia Everwin Gardens Rm 502 Block B 521 Wanping Nan Lu Shanghai 200030 China Tel 86 0 21 64869685 Fax 86 0 21 64280591 www dmt asia com sales dmt asia com Support dmt asia com DMT USA Inc 1201 Peachtree Street 400 Colony Square Suite 200 630 Atlanta GA 30361 USA Tel 1 770 612 8014 Fax 1 678 302 7013 www dmt usa com
42. erence Setting for Chamber 1 must be O 8 Chamber 3 0 8 Chamber 4 0 9 3 2 3 Analog output Setup gt 750TOBS parameters gt Analog output Analog Output The output is from 0 0 to 2 5 Volt Resolution is 0 61 mV In this window the span can be adjusted LO Vol 10 m In the example above the span is set to 110 mN The output is O O Volt if the force is 10 mN and 2 5 Volt if the force is 2 5 Yolk 100 mM 100 mN Direct Output The internal sample frequency is 20 Hz on each channel Any ou digital filtering in your data acquisition software must beset f above 20 Hz If the setting Direct output is choosen the settings above is not used The output is then routed directly from the transducer Offset 2 5 Volt Output 300UV mN If Direct output is used the signal is not filtered and the output is only limited by the transducers response time it is approximately 10 mS 3 2 3 8 Weight calibration data Weigth Calibration Data Setup gt 750TOBS parameters gt Weight cali bration data A read out of the Force calibration data Only Chop for internal use if there are some problems with a transducer Calibration Data Zera 093 Gain Chamber 2 hogi Chamber 3 2216 Chamber 4 5447 User manual 750TOBS Chapter 3 23 3 2 3 9 Wash Setup gt 750TOBS parameters gt Wash Wach p sec every f min E times Programming of the wash sequence M n
43. ext wash The wash sequence works b overflow Wash E Sec every f min fs times In the example to the right the wash sequence is set to Wash 4 sec every 1 min 2 times then wash 3 sec every 1 min 5 times If next wash is not checked there will only be one wash Wash Setup 3 2 3 10 Flow calibration Setup gt 750TOBS parameters gt Flow calibration Before this calibration is performed the low flow shunt supplied in the calibration kit must be mounted on the tubing inside the reservoir bottle Calibration of flow in Constant Wash mode is performed by the marked Tissue chamber provided with the calibration kit 5ml Optional for the 750TOBS please contact DMT for purchase On the Tissue chamber there are two marks used for the calibration The calibration is done one chamber at the time 1 Turn off oxygenation Sei 2 Choose the chamber to calibrate Click Start flow ET E Re O sec Cancel 3 When the surface of the fluid reaches the bottom mark on the calibration tissue D chamber click the Start Measure button Check this by looking at the line at eye level and look for the bottom of the fluid meniscus to reach the line Start Measure 4 When the surface of the fluid reaches SS Flow Calibration tj the top mark on the calibration tissue I chamber click the Stop Measure am button Stop Measure Cancel DMT Tissue organ bath system Model 50TOBS 5
44. force is zeroed or when a drug is added A dashed line on the datasheet indicates where a comment has been added The dashed line can be made invisible by removing the check mark next to Comments in the View dropdown menu To add a comment Choose which channel the comment has to belong to Click on the Add button choose a default comment or write a new comment in the text box Important Comments can only be added when acquisition is started Click on Add to save a comment e Setup View Wash About amp amp hd QQ 4 44 4 P P A 20956 se Ql k OP E ni ita View Comments Data Pad De uM Phenylepheine e Tissue Chamber 1 qo Zu ag Fy hes FR 50 0 2 i Force 23 4 i Auto Al M Zero Tissue Chamber 2 e Ba E Auto E Zero Tissue Chamber 3 T Us Temp aT Force 4 Auto c Zero Tissue Chamber 4 ag Temp EN Force 4 Auto E Zero D 09 37 00 08 38 00 08 39 00 08 40 00 03 41 00 09 42 06 09 43 00 09 44 00 Fa Choose which channel to Write or choose a default comment on O all channels comment e Double clicking on the comment input field opens the Comments Setup 20 default comments can be stored here SS G m QYyZLLQQQ Q111 LL LLL LIIS Comments Setup DMT Tissue organ bath system Model 50TOBS 3 5 4 Viewing comments
45. hat can be opened in an Excel Worksheet TY 750TOBS Software A File Setup View Wash About Mew Operi Save Save Data view bo File Export Data File to Excel Worksheet Export Data Pad to Excel worksheet Export ke Print Auto Backup Exil f Jan Start Start a View bo Clipboard Open the 750 file in a Microsoft Excel worksheet E File Edit view Insert Format Tools Data Window Help D Ge Ej sey elo 4 BA J ve 417 Wy aime 2 Arial sho plezu ebe 58 EH Nr A i ta fa fa a Ch a 2 Rei GA Y Renly with changes End Review 116 fe preme mE jExperiment Age Sex Comment 1 Comment 2 Comment 3 Comment 4 Comment 5 Comment B Comment 7 11 Comment 8 12 Comment a 13 Comment 10 uti CA LI In m HE GE rm Fa tu NER Lig On Ao E CO RO 15 Comment Chamber Max Min Mean SID DEM samples Start time End time 17 Start 14 1182 1 2348 B 2105 4526 13646 11 30107 41 39107 41 15 Zero force 16 9215 0 0917 5 1053 45772 0 4425 107 39107 41 39107 41 1 2 19 Zero force 3 185117 0 0408 778311 7 5155 10848 48 38107 41 39107 41 20 Start 2 169419 00306 81507 83488 2 4101 12 29107 A1 39107 41 1 1 1 21 Start 14 1182 0 0917 6 0394 5 3423 1 3794 15 39107 41 38107 41 00917 01019 00943 00051 0 00245 4 39107 41 38107 41 0 0917 z BSTA 0 0917 DU uU 11 38107 41 38107 41 User manual 750TOB
46. ia uer en tack uen ege Eege d e aere 32 3 6 2 Export a Datapad file to an Excel worksheet ceeeeeeerreeeene 33 3 6 3 Export a Data file to an Excel worksheet eee nnns 34 EEN 35 SE FS EE 39 User manual 750TOBS Contents 10 Chapter 4 The Tissue organ bath unit sss 36 4 1 The first weight calibration 1e eere rene 36 4 2 Calibration of force transducer KEE rennen nnn 36 4 3 Tissue chamber handling 1e eere erre nnn 37 Ao Ad sing te eege 36 4 5 Tissue organ bath maintenance eere eere nennen 40 4 6 Tissue organ bath cleaning KEREN KEREN KEREN KEREN KEE ERE 40 Chapter 5 Tissue organ bath accessories and spare parts 41 5 1 General Tissue organ bath equipment eene 41 5 2 Tissue organ bath system accessories ENNEN K ENKEN Ek 42 Chapter 6 Terms of warranty 1 11e eeeeee eene nnne nau 43 Appendix 1 Service check eeieeeee eere KEREN KEREN KEREN KEREN d Appendix 2 Shipping instructions eere res 45 Appendix 3 System specifications eere enn 46 Appendix 4 Fuse chang
47. ing eesecseeee rere ener nnn nnn nnn nnns 47 Appendix 5 Software installation eren nnn AS Appendix 6 VCP Driver installation esee 50 Appendix 7 Buffer recipes 1 nece eere rennen nnn KEE KENER KEREN nn auunn 53 7 1 Buffer recipes E 53 7 1 2 Physiological saline solution PSS cc secsssescssseuccsseseccsesecceneucnsnenccessseccsenscnens 53 E BA E 55 Appendix 8 750TOBS controller eere eee nenne 57 8 1 The 750TOBS Software User Interface 57 ENEE 7 OTOD RERO 57 e 2 TIS wicdoW a a E N 57 92 2 DOD GOW ii E NUS E 58 ey SI MENU EE 58 8 2 2 2 SIE Engler le TE EE 59 92 2 EEN 59 8 2 2 4 Heating plate temperature secceccsccedennnenctacanastecansedtenaanduncandsaaeenceadawassienceusndeinens 59 8 2 2 5 Temperature GIffErENCe cccsccssseccceesccseseneessnseusnnsenceseneeeenausenensenceeenanseneeass 60 8 2 2 ST ANG s 972 OUTPUT RR ot a eer TTC 60 8 2 2 7 Weight calibration data seduta Ga bomcetsaunecctoataremtceanmecetsuanetlGazenuneatuacieeubatncabs 60 RPA i A E E E E NE E E E E 61 EEN FONC a O DEE 61 8 2 2 10 Mode EE EE 62 Bor Weight er ele ON usn ERa E EEEE EEES 64 OL VIO 65 EA SIN ASM aa 65 SL E EE 66 E E EE 66 DMT
48. ints the Data pad on the default printer DMT Tissue organ bath system Model 50TOBS 3 2 3 2 Setup menu Connect to 750TOBS e Connect or disconnect the 5OTOBS 450TOBS parameters e See sections 3 2 3 3 to 32 942 Chamber 1 T 5nal amp Max value 44 7 Min value Chambers The window can also appear by double clicking on the Y axis Y axis setting e Y axis color Tempo Set Temp 37 0 20 mil Volume SCH D 750TOBS Software A i File Setup View Wash About ser 2 7 eForce units Choose between mN or gram ps FSOTOBS paramaters P Te Connect to 7SOTOBS j e Chamber 1 Chamber 2 Chamber 3 Chamber 4 e Axis amp Grid e Comments i Force Units K i F User InterFace d Axis and Grid Ac Format MM SS 222 If AIP is checked the changes will be applied to all four chambers In Constant Wash Mode the Volume setting is changed to Flow and units to ml min Comments Setup e dd hhimmess mm dethh mm ss User Interface There are two i choices 1 Controller described in Appendix 8 It is used when another software package is used for acquisition 2 Acquisition amp Controller C arid M Show Som Iw Show od j Color um Iw Show t anis Axis and Grid The window can also A appear by double clicking on the X axis i Change the settings for Set Temp i and
49. ion procedure is described in detail in Section 3 2 2 12 Figure 4 1 Transducer As a part of the general maintenance of the Tissue organ bath system DMT recommends that the organ bath is weight calibrated at least once every month DMT also recommends that the bath is weight calibrated every time the system has been moved or has not been used for a long period of time DMT Tissue organ bath system Model 50TOBS 4 3 Tissue chamber handling The following is a description of how to handle the tissue chambers and the different kinds of sample holders available Placing the glass 1 Move the transducer upwards Make sure the sample holder is free of the tissue chamber 3 Place the pipe stub at the bottum of the chamber inside the chamber disc Gently place the pipe stub situated on the side inside the heating compartment Removing the glass Carefully pull the top of the chamber outwards a few millimeters until the chamber is free to move upwards User manual 750TOBS Chapter 4 37 4 3 1 Adjusting the transducer When tissue is placed in the sampleholder do a course adjustment on the tissue holder to fit the tissue just between the holder and the transducer without stretching it With the tissue submerged in the buffer set the force to 0 Then use the micrometer positioner to stretch the tissue to the disired resting tension fee 7 Horizontal adjustment of N A RN the transducer bos N Tur
50. ions pulmonary arterial smooth muscle and even lung parenchyma Other smooth muscle preparations that are used in organ bath research are urinary bladder penile corpus cavernosum muscle strips and prostate Because of these inherent qualities and the versatility in use the basic organ bath design has seen little change over the last few decades Nonetheless there are some practical drawbacks e g they take much bench space use circulating water by causing temperature delays and gradients are tough to clean hard to automate and non GLP compliant To remedy these limitations DMT has now redesigned the organ bath system for the new millennium using the latest developments in material computer and tissue engineering We took away the heating by water replaced it by very fast air heating made the chambers disposable or reusable after cleaning added automated control of filling emptying and gassing by an embedded microprocessor and provide digital output of force and complete computer control of the instrument All of this in a box that takes up a quarter of the space of a traditional system The modular design allows up to four individually mounted tissues to be studied simultaneously and independently per system Moreover multiple systems can be linked together USB connection for high throughput screening HTS and up to 16 units can easily be run by a single person The convenient compact bench top design of the DMT s organ bath syste
51. is time Click Next to continue Found New Hardware Wizard This wizard helps you install software for USB lt gt Serial Cable If your hardware came with an installation CD lt 6 or floppy disk insert it now What do you want the wizard to do Install the software automatically Recommended Install from a list or specific location Advanced Click Next to continue User manual 750TOBS Cancel Appendix 6 49 Select Search for the best driver in these locations and enter the file path in the combo box C VCP Drivers in the example below or browse to it by clicking the browse button Once the file path has been entered in the box click next to proceed If Windows XP is configured to warn when unsigned non WHQL certified drivers are about to be installed the following screen will be displayed Click on Continue Anyway to continue with the installation If Windows XP is configured to ignore file signature warnings no message will appear The following screen will be displayed as Windows XP copies the required driver files Found New Hardware Wizard Please choose your search and installation options ae S G Search for the best driver in these locations Use the check boxes below to limit or expand the default search which includes local paths and removable media The best driver found will be installed C Search removable media floppy CD R
52. l Calibration of each chamber is performed with the marked tissue chamber provided with the calibra Chamber 4 280 m5ec ml 350 mec jml tion kit 20 ml Chamber 3 200 m er md 350 msec jml Adjust the numbers of each tissue chamber until the amount of fluid is correct the numbers indicates msec ml If the amount of fluid is to small the number msec ml must be higher If the amount of fluid is to high the number msec ml must be lower 8 2 2 3 Pressure Setup gt 750TOBS parameters gt Pressure Adjustment and on line reading of pressure This set ting is normally not changed Actual pressure Act Pressure 0 5 Act Pressure provides an on line reading of the ac tual pressure in the reservoir bottles Sek Pressure p 400 Set pressure Update To set or reset the target pressure 0 01 0 999 Bar Default 0 400 Update Update provides an on line reading of the actual pressure NOTE When the pressure is reset press the small button that appears to the right of the Set Pressure Input field If the pressure is changed the flow must be calibrated again as described in paragraph 3 2 3 10 8 2 2 4 Heating plate temperature Setup 750TOBS parameters gt Heating plate temperature Adjusts the temperature on the heating plate on which the reservoir bottles are placed The temperature here is chosen to be higher than the bath temperature to avoid gas formation in the lines and to corre
53. l chambers or an appropriate chamber will be drained by activating the Fill amp Empty Control see 3 2 2 When using a vacuum pump use around 11 L min NOTE Please be aware of local regulations and codes for hookup to house vacuum systems A VacuShield is recommended and local regulations may require shielded non glass Nalgene bottles and reservoirs to be used to collect waste NOTE When draining the chambers using the automatic suction function please press the appropriate Fill amp Empty control for an additional 3 5 sec after the chamber appears to be empty This will prevent leftovers from used solutions being retained in the tubing and valves User manual 750TOBS Chapter 2 15 16 Chapter 3 User software Chapter 3 is a complete manual for the Tissue organ bath Data acquisition software The chapter contains a step by step description of how to navigate in the software and how to use the special features of the Tissue organ bath system If the user only wishes to use the 750TOBS Controller program please see Appendix 8 p 57 A software installation guide can be found in Appendix 5 3 1 The Software program The 750TOBS Software is generic to the Tissue organ bath application and gives the software a particular flexibility which users may find helpful in organizing their work The software is used to control and acquire data from the 75OTOBS The 750TOBS Software provides many ways of displaying data The grid
54. le Go to the end one page at a time Go to the end 1 10 page at a time Undo up to the last 10 zoom actions Start Stop recording Choose sample rate a 9 dd O Resume to the end of the datasheet Scroll Zoom Select Activate to select comments in the dataview To view a comment hold the cursor above a comment line left click and a new window will open showing data from the selected comment Cursor The cursor is moved by holding the left mouse button while moving the mouse There are several options for reading data Right clicking on the cursor opens an op tions menu Full screen Upper half when 2 programs are open at the same time Lower half when 2 programs are open at the same time User Manual 750TOBS Chapter 3 17 Zoom selection of data in the Data view An easy way of selecting data in the Data View is by dragging the mouse over the area while holding down the Alt key X axis and the left mouse button or the shift key Y axis in the one channel view only or the Alt shift X and Y axis in the one channel view only Zoom in on the X axis Four and one channel view Zoom in on the Y axis One channel view only Zoom in on the X axis and Y axis One channel view only T 750TOBS Software File Setup View Wash About TQ Q bl QQ KK gt A FE 3 Q kok De TSOTOBS A Comments Test tab Data Pad TZ el Alt left mouse button YL T 2223 13 30 00 13 35 00
55. m can be placed in any existing laboratory environment while gaining back valuable bench top space DMT Tissue organ bath system Model 50TOBS Safety DMT Tissue organ bath system 750TOBS has been designed for use only in teaching and research applications It is not intended for clinical or critical life care use and should never be used for these purposes nor for the prevention diagnosis curing treatment or alleviation of disease injury or handicap e Do not open the unit the internal electronics pose a risk of electric shock e Do not use this apparatus near water e To reduce the risk of fire or electric shock do not expose this apparatus to rain or moisture Objects filled with open liquids should not be placed on the apparatus e Do not block any ventilation openings Install in accordance with the manufacturer s instructions e Only use secure industry standard connectors and tubing for pressure and vacuum connections Faults defects and mistakes due to wrong connections void warranty We are not accountable for results and mistakes due to inappropriate pressure or vacuum hookup e Do not install near any heat sources such as radiators heat registers stoves or other apparatus that produce heat e Only use attachments and accessories specified by the manufacturer e Unplug this apparatus during lightning storms or when unused for long periods of time e This apparatus must be grounded e Use a three wire grounding
56. methods and based on own judgment determine what is best for the experimental conditions When the Constant wash mode is selected go to Setup gt Chamber 1 4 or double click on the chamber s y axis to adjust the settings T3 T50TOBS Software A File Setup View Wash About D Connect to 750TOB5 gt dd d b bb B 20 s sec Start 9 e 75 TOBS paramaters r T Chamber 1 KR P d I om Chamber 2 Chamber 3 i Chamber 4 i g Axis amp Grid PIRE I RU ELA Lampe cena enc eat e Et Eu E ETE Comments i Force Units 0 0 1 n Chamber 1 Scale r Temp Val i May value 50 WEE TORE DAE H Min value LN Flow 0 5 TUUM DMT Tissue organ bath system Model 50TOBS Important When the mode is changed the shunt inside the bottle must be changed too The Constant wash mode requires the Constant flow shunt and Fill Empty mode requires the Fill Empty flow shunt Important Make sure that the pressure is OFF before changing the shunts A warning noting that the pressure is turned off is Attention shown when the mode is changed You are changing mode Please check that the right shunt is mounted in the bottle Please note that the pressure is removed Constant flow shunt Hypodermic needle 0 4 x 40mm Lc nn wem Fill Empty flow shunt Accessories please contact DMT for purchase User manual 750TOBS Chapter 3 27 3
57. n the micrometer screw clockwise to unlock the transducer When unlocked it is possible to move the transducer horizontially With the other hand Turn counter clockwise to lock oe c A f Course adjustment of the lower tissue holder Turn the micrometer screw clockwise to unlock the transducer When unlocked it is possible to move the tissue holder up down left and right Turn counter clockwise to lock Vertical fine adjustment and stretching the tissue Turn the micrometer screw clockwise to lower the transducer Turn anti clockwise to increase the distance The marked line is the lowest position of the transducer The highest position is 20mm above the line 38 DMT Tissue organ bath system Model 750TOBS 4 4 Sample holders Figure 4 4 1 Triangle hooks Figure 4 4 2 Closing clips optional Figure 4 4 4 Triangle hooks w Figure 4 4 5 Triangle hooks w circular stimulation electrodes optional stimulation electrodes optional User manual 750TOBS Chapter 4 39 4 5 Tissue organ bath maintenance The Tissue organ bath system is a very delicate and sophisticated piece of research equipment In order to keep it working at its best DMT recommend that the following sections are read carefully and that the instructions are followed at all times Important It is VERY important that the tubing being used is the original tubing from DMT The quality diameter and length a
58. nel Mounted to the lid inside the bottles A constant flow shunt or a fill empty shunt en p pentes e Micrometer positioner Perfusion buffer inlet e Transducer e Tissue chamber ETC ee 3 Power indicator Sample holder T 3 Lo Oxygenation CO regulator Chamber door e ee Glass presenec indicator Figure 1 1 Tissue organ bath front panel DMT Tissue organ bath system Model 50TOBS 1 2 Tissue organ bath rear panel OXYGEN MAX 10 BAR vaccum e EM EE oo LEVEL RST USB REC A REC 3 REC 2 REC 1 Communication indicator 115 230V 50 60Hz e P0 Po iP automatic voltage selector USB connection ois toten P1 Analogue recorder output e mM NE WM ON OFF CAME E le Suction connection for vacuum pump Pressure inlet max 10bar 145psi Figure 1 2 Tissue organ bath rear panel User manual 750TOBS Chapter 1 13 1 3 Tissue organ bath unit e Chamber detector eee e Tissue chamber Air heating C Sample holder lower part shown Figure 1 3 Tissue organ bath unit NOTE The chamber detector registers when the tissue chamber is placed If the chamber is placed correctly the blue indicator lamp will be lit and the Tissue organ bath will work ve e Knob for dismounting chamber disc and temperature probe Glass indicator e EE e gt e Oxygenation On Glass mounted CO regulator Off Glass removed Temperature probe e
59. owing product DMT Tissue Organ Bath System Model 75O0TOBS Version 2 2 is covered by this certificate and marked with CE label conforms with the following standards EN 61010 1 2001 Safety requirements for electrical equipment for EN61010 1 Corr 1 2003 measurement control and laboratory use EN 61010 1 Corr 1 2003 Part 1 General requirements EN 61010 2 101 2003 Safety requirements for electrical equipment for measurement control and laboratory use Part 2 101 Particular requirements for in vitro diagnostic IVD medical equipment EN 61326 2 6 2005 Electrical equipment for measurement control and laboratory use EMC requirements Part 2 6 Particular requirements In vitro diagnostic IVD medical equipment With reference to regulations in the following direc tives 2006 95 EC 89 336 EEC User manual 750TOBS Certificate of Conformity About this manual This manual contains a complete list of procedures describing how to install maintain and get started using the Tissue organ bath system Model 750TOBS Version 2 2 Chapter 1 provides an overview of the construction and basic features of the Tissue organ bath system Chapter 2 describes step by step how to setup a complete Tissue organ bath 7SOTOBS system including accessories Chapter 3 is a complete manual to the Interface The chapter describes in detail the construction of the software and how to use all the features of the Tissue organ bath system
60. ply with the limits for a Class B Digital device pursuant to part 15 of the FCC rules These limits are designed to provide reasonable protection against harmful interference in residential installations This equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the instructions may cause harmful interference to radio communications However there is no guarantee that interference will not occur in a particular installation If this equipment does cause harmful interference to radio or television reception which can be determined by monitoring the interference while turning the equipment off and on the user is encouraged to correct the interference by one or more of the following measures e Reorient or relocate the receiving antenna e Increase the separation between the equipment and receiver e Connect the equipment into an outlet on a circuit different to that which the receiver is connected to e Consult the dealer or an experienced radio TV technician for help Approvals Complies with the EMC standards EMC 89 336 EEC EN 61326 2 6 2005 EN 61000 3 2 Certified with the safety standards Directive 2006 95 EC EN 61010 1 2001 EN 61010 1 Corr 1 2003 EN 61010 1 Corr 1 2003 EN 61010 2 101 2003 DMT Tissue organ bath system Model 50TOBS Certificate of Conformity DMT A S Skejbyparken 152 8200 Aarhus N Denmark hereby declares its responsibility that the foll
61. r SE ete EG T Connectto750TOBS e Tissue Chamber 2 a bath and refilling it with FSOTOBRS paramaters d Fil Empty calibration emp 36 B fresh buffer via a Fill empty f chamber Pressure m Chamber 2 Heating Plate temperature On EENG The adva ntage B Chamber 3 Temperature Difference Ls Zero Gel Fill i of this method IS that the Vos Chamber 4 Analog Output concentration of drug Force Units Weight Calibration Data User Interface k Wash declining fast especially if M Ho Calibration this is repeated 3 times or Made K Constant Wash more The 750TOBS can do Calibrate Transducers boy this automatically via the wash menu and the buffer comes from the pre heated buffer reservoir in the system Therefore compared to the classic approach washing is easy convenient and fast A disadvantage of this fill empty method is that the tissue is briefly suspended in air without nutrients and stimulation when used needs to be suspended When the fill empty mode is selected go to Setup Chamber 1 4 to adjust the settings 13 750TOBS Controller A File Setup Views Wash aa Connect to T5 TOBS 75 TOBS paramaters zs 1 al Charfiter 2 e Wi Chamber 3 Chamber 4 r Chamber 1 r Y 5 cale r Temp al Force Units 50 37 0 User InterFace Mas value Set Temp Min value D Volume 20 ml E AI Wm A User manual 750TOBS Appendix 8 62
62. r computer To continue click Next WARNING This program is protected by copyright law and international treaties Click Next T50TOBS InstallShield Wizard License Agreement Please read the Following license agreement carefully By installing copying accessing or using the SOFTWARE or its OUTPUT you agree to be bound by the terms of this license agreement IF you do not agree to the terms of this license agreement then DO NOT install or use the SOFTWARE THIS SOFTWARE IS NOT FOR SALE The SOFTWARE is licensed to you as the end user itis not sold Copyright Notice All content copyright Danish Myo Technology 4 5 2006 all rights reserved I accept the terms in the license agreement OI do not accept the terms in the license agreement Installshield Accept the terms and click Next Ready to Install the Program The wizard is ready to begin installation If you want to review or change any of your installation settings click Back Click Cancel to exit the wizard Current Settings Setup Type Typical Destination Folder C Program FilesDMTY750TOBS User Information Name Companv Installshield The program is ready to be installed click Install User manual 750TOBS Appendix 5 3E 750TOBS InstallShield Wizard B 47 48 T50TOBS InstallShield Wizard Installing 750TOBS The program Features you selected are being
63. r installing the serial converter driver Now the 50TOBS Controller Software can be opend and connected to bOTOBS as described in paragraph 3 2 1 Found New Hardware Wizard Completing the Found New Hardware Wizard The wizard has finished installing the software for USB Serial Converter Click Finish to close the wizard Finish User manual 750TOBS Appendix 6 51 Appendix 7 Buffer recipes 7 1 Buffer recipes NOTE The Schott Duran bottles should not be filled up with more than 800 ml of solution NOTE To minimize the heating time one can benefit by preheating the buffer to 35 C before filling the reservoir bottles 7 1 2 Physiological Saline Solution PSS 1x PSS 1000ml Solution 1 Chemical MW g mol Conc mmol l Conc g l NaCl 58 44 118 99 6 954 KCI 14 56 4 69 0 350 MgSO H O 246 48 1 17 0 289 KH PO 136 09 1 18 0 161 Solution 2 Chemical MW g mol Conc mmol l Conc g l CaCl 2H O 147 02 2 50 0 368 Solution 3 Chemical MW g mol Conc mmol l Conc g l NaHCO 84 01 25 00 2 100 EDTA 3 2 24 0 03 0 010 Glucose 198 77 5 50 1 091 1 Dissolve the chemicals in 100 ml double distilled H O as three individual solutions as described in the table above Gently heat solution 3 to dissolve the EDTA 2 Solution 1 is added to a graduated bottle and the bottle is filled with double distilled H O to a final volume of 500 ml 3 Solution 3 is added to the graduated bottl
64. re VERY important in order to insure the correct draining and filling of the chamber 4 6 Tissue organ bath cleaning DMT strongly recommends that the tissue bath chamber and surroundings should be cleaned after each experiment After a normal experiment use the following procedure to clean the tissue bath chamber tubing and tissue holders 1 Turn Pressure OFF 2 Useclean Schott Duran bottles 3 Fill with distilled water 4 Turn Pressure ON Figure 4 6 1 5 Rinse the silicone tubing with double distilled water by Schott Duran bottle activating the Fill amp Empty Control several times SC Temperature probe NOTE Only works when a tissue chamber is in place p i 6 Change the tubing after use of unknown or toxic chemicals or if any kind of hydrophobic reagent has been used f Carefully dismount the tissue bath chamber and perform a normal glass wash 8 If aggressive solution or drugs have been used it is possible Figure 4 6 1a to disconnect the temperature probe Fig 4 6 1a and take Chamber disc and out the chamber disc Clean both with double distilled water MP probe and dry with a paper towel Important Note In exceptional cases it may be necessary to demount the sample holders for cleaning to make sure that all surfaces are cleaned DMT Tissue organ bath system Model 50TOBS Chapter 5 Tissue organ bath accessories and spare parts This chapter contains a complete list of equipment needed
65. type cord similar to the one supplied with the product e Do not defeat the safety purpose of the polarized or grounding type plug A polarized plug has two flat blades one being wider than the other A grounding type plug has two blades and a third round grounding pin The wide blade or the third prong is provided for your safety If the provided plug does not fit into your outlet consult an electrician for replacement of the obsolete outlet Be advised that different operating voltages require the use of different types of line cord and attachment plugs Check the voltage in your area and use the correct type See the table below Voltage Line plug according to standard 110 125V UL81 and CSA C22 2 No 42 220 230 V CEE page VII SR section 107 2 D1 IEC 83 page C4 240 V BS 1363 of 1984 Specification for 13A fused plugs and Switched and unswitched socket outlets Protect the power cord from being walked on or pinched particularly at power plugs and the point where they connect to the apparatus Refer all servicing to qualified service personnel Servicing is required when the apparatus has been damaged in any way such as the power supply cord or plug is damaged liquid has spilled onto or objects have fallen into the apparatus the apparatus has been exposed to rain or moisture does not operate normally or has been dropped User manual 750TOBS Safety EMC EMI This equipment has been tested and found to com
66. w If the connection fails the message display is Not connected In case the connection fails try connecting by clicking on Setup Connect If the connection fails again check if the USB cable is mounted correctly and 5OTOBS is turned on If two bOTOBS are connected to one PC use Ho connection to SU0TOBS The following COM ports are available Check that the USB cable is connected and the power is turned on both programs Always connect 750TOBS r m Software A before 750TOBS Software B 8 2 Control windows 8 2 1 The Main window e Drop down menus 13 750TOBS Controller A File Setup View Wash About Register gt Tissue Chamber 1 ss Tissue Chamber 2 Temp E F a Ta Temp S Force hi DU re Zero Fil eS E zen Fill d P eee Seseeee ee Seeeeeee 99006005 Sseeeeee 59026005 IL eee Figure A8 1 The Tissue organ bath User Interface DMT Tissue organ bath system Model 50TOBS Tissue Chamber 3 as RR p Tissue Chamber 4 Temp 97 AT Ta Force 0 0 y Zem Fill Auto Mode Fill Empty e Statusbar Temp P B n Force i a E Zen il ET Tissue chambers controls e Fill chamber Empty chamber x o Las e Pressure is activated deactivated by clicking the off e button below or e ef sese D ee ee Tissue Chamber Ee Temp E 6 7 a If Auto is
67. website 13 750TOBS Software A File Setup View Wash About Regisf amp r IRAMA 4 44 4 IMMENSE Data View Comments Data Pad Tissue Chamber 1 8 Ta Temp 3 i Force 0 0 ET Auto VY Al sn E Tissue Chamber 2 Force 0 0 po H Au s Zero Tissue Chamber 3 4 Us n 53 M Force 0 0 zi zen iE Se Tissue Chamber 4 pes AT By Temp G Force 0 0 16 10 00 Auto a Zen F Connected to COMS File C ADMTIOrganbadlAKUN datafilerl zMarchz007107March2007 det Chamber 1 4 Mode Z User manual 750TOBS Chapter 3 35 36 Chapter 4 The Tissue organ bath unit Chapter 4 contains a complete explanation of how to calibrate and maintain the Tissue Organ Bath units to ensure the equipment is always performing to the highest standard 4 1 The first weight calibration Prior to the shipment of the Tissue organ bath 50TOBS system it has gone through two days of continuous testing including a final weight calibration However in order to ensure that the Tissue organ bath is working at highest performance DMT recommends that a new weight calibration is performed before starting to use the Tissue organ bath system The calibration is performed through the calibration menu using the appropriate weight according to gain setting As default the system is set with a gain range of 20 200 mN and the calibration is performed using a 5g weight fig 4 1 The weight calibrat
68. z on each channel Any E digital filtering in your data acquisition software must beset f above 20Hz If the setting Direct output is choosen the settings above is not used The output is then routed directly from the transducer Offset 2 5 Volt Output 300 uV mN If Direct output is used the signal is not filtered and the output is only limited by the transducers response time it is approximately 10 mS 8 2 2 7 Weight calibration data Weigth Calibration Data Setup gt 750TOBS parameters gt Weight cali bration data A read out of the Force calibration data Only CES for internal use if there are some problems with a transducer Calibration Data Zera 093 Chamber 2 hogi Chamber 3 216 Chamber 4 2447 User manual 750TOBS Appendix 8 59 8 2 2 8 Wash Wash Setup Setup gt 750TOBS parameters gt Wash Bo I am Wash d sec every 1 mint 2 times Programming of the wash sequence jw next wash The wash sequence works by E 1 in 5 i overflow Wash sec every mir times In the example to the right the wash sequence is set to Wash 4 sec every 1 min 2 times then wash 3 sec every 1 min 5 times If next wash is not checked there will only be one wash 8 2 2 9 Flow Calibration Setup gt 750TOBS parameters gt Flow calibration Before this calibration is performed the low flow shunt supplied in the calibration kit must be mounted on th

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