SOF-700RA-29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE
Contents
1. D I 0 12 15 18 21 24 27 30 0 000 0 000 5 000 3 000 30 000 2 400 30 000 30 000 000 000 000 000 000 000 000 000 000 000 000 30 3 180 60 151 340 470 720 710 140 980 060 920 860 450 000 000 000 000 000 789 100 1 000 0 000 0 100 0 000 5 628 460 200 220 890 340 410 900 040 860 780 560 10 Named Variables Array Defaults Acclimation Period 5 minutes Number of Trials to 30 Start Safe Side 1 Left Stimuli on Safe Side 3 Red Stimuli on Shock Side 3 Red Stimuli Flash Rate 0 5 seconds Delay to Shock Onset on Dark Side 0 seconds Max Shock Duration 30 seconds No Response Limited Hold 180 seconds Shock On Time 0 1 seconds Shock Off Time 2 4 seconds Session Time 60 minutes Summary Data Array Crossings Escapes Latency Trial by Trial Array D I Trial Number 0 1 1 Latency to Cross into Shock Zone No Response Time default 180 seconds if no Crossing 0 1 2 Latency to Escape from Shock Maximum Shock Duration default 30 seconds if no Escape Rev 1 0 Copyright O 2012 MED Associates Inc 5 Data Analysis Using MED PC to Excel Using a Pre Formatted Table Profile MTP file MED PC to Excel MPC2XL is a program2. Acclimation 0 51 14 27 2012 4 22 29 PM Rev 1 0 Copyright O 2012 MED Associates Inc MED ASSOCIATES INC SOF 700RA 29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE Viewing Changing Variable Values A protocol s variables may be changed from their default values even once a session is in progress Change variables by selecting Configure Change Variables on the main menu or click the 4th tool bar item AX In the Display Data from Box section located in the lower left hand corner of the Displaying Variables window Figure 3 5 choose the chamber s to modify By clicking additional boxes in the Additional Boxes to Update section changes made to a single box are duplicated on the boxes selected Figure 3 5 Displaying Variables Screen Displaying Variables from Box 1 A Array B Array ao D Array RS 2 Named Vars G H J 5 Vars 0 000 0 000 0 000 0 000 0 000 0 000 0 000 0 000 0 000 0 000 Befresh Q T 0 000 0 000 0 000 0 000 0 000 Issue U X Y E 0 000 0 000 0 000 0 000 0 000 ose 2 x 0 000 d Help Display Data from Box Additional Boxes to Update C from Box 1 p me 0 000 t Select All Deselect All In the Displaying Variables screen click an array on the table and each element in that array can be viewed To change a value select the variable in the upper left section of the dialog and replace the value in the lower right hand box or
3. MED ASSOCIATES INC SOF 700RA 29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE CHAPTER 1 Introduction MED PC gives researchers the ability to use pre programmed applications such as the Zebrafish Shuttle Box Avoidance to make data collection easy These pre programmed applications can also be modified to meet the evolving demands of a research protocol This manual provides step by step instructions on how to use this pre programmed application In addition to this manual refer to DOC 010 MED PC IV User s Manual for the installation of the MED Associates interface drivers the MED PC IV Software and the Delphi Compiler Also refer to the Wiring Guide that accompanies the hardware for instructions on hardware configuration The Hardware Configuration software utility is installed with MED PC IV and is used to assign the inputs and outputs in the interface cabinet for each task controlled by the program Figure 1 1 shows the ENV 012Z X1 Zebrafish Shuttle Box for which this protocol was developed Figure 1 1 ENV 012Z X1 Zebrafish Shuttle Box Data file structure file saving format and other related options are also determined by the Hardware Configuration software utility Refer to DOC 003 MED PC IV Programmer s Manual if you are unfamiliar with the process of translating and compiling an application The staff at Med Associates Inc is available to answer any questions that may arise Please contact us via e mail at support med associa
4. amp Understanding the Raw Data File Unless otherwise specified data will be saved to MED PC installation directory default is C MED PC IV Data The data file format is specified in the Hardware Configuration Utility See DOC 010 MED PC User s Manual for data file format options Data are saved at the end of the protocol or can be saved manually by selecting File Save Data Manually or File Save Data Flush Data files may be opened with Notepad Wordpad or any word processor spreadsheet software however ensure they are always saved unformatted for use with data extraction utilities such as MED PC to Excel MPC2XL Data file formats are explained in detail in the MED PC IV User s Manual File C MED PC 2012 04 25 16h18m Subject 0 Start Date 04 25 12 End Date 04 25 12 Subject 0 Experiment 0 Session identification information entered when starting the Group 9 experiment Box 1 Start Time 16 18 45 End Time 16 27 41 MSN Zebrafish Shuttle Box Passive Avoidance Training 0 000 000 000 000 0 0 0 0 000 Subscript for Data Array D H cr m m oo M 000 c 000 L Location Flag 1 Left 2 Right 000 000 000 000 000 000 000 000 194 000 000 000 000 000 000 S Acclimation Session Timer O P O x Rev 1 0 Copyright 2012 MED Associates Inc
5. that helps to import data from MED PC the raw data file format previous section to a spreadsheet program such as Microsoft Excel MPC2XL needs to be installed separately from MED PC IV Please refer to DOC 036 User s Manual for MPC2XL for installation instructions Follow the step by step instructions below for importing data obtained from the Zebrafish Shuttle Box Passive Avoidance mpc procedure Open Microsoft Excel and select cell A1 Minimize the Excel window Open MED PC to Excel The MED PC to Excel window will be displayed as shown in the Figure 5 1 In the Transfer Data window located in the leftmost file tab click Select in the Table Transfer section at the bottom of the dialog window Figure 5 1 Select Table Transfer Profile mtp file MED PC to Excel Bi File Help Transfer Data Edit Row Profiles Edit Table Profiles Row Transfer Profile NO PROFILE ASSIGNED Transfer Orientation Column Labels Horizontal Rows Data C Vertical Columns Table Transfer Profile NO PROFILE ASSIGNED Transfer Labels Choose Zebrafish Shuttle Box Passive Avoidance Training mtp in the C MED PC IVAData folder 11 Rev 1 0 Copyright O 2012 MED Associates Inc MED ASSOCIATES INC SOF 700RA 29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE Figure 5 2 Click Transfer to Select Data file to Transfer to Excel MED PC to E
6. use the up down arrows to increment by 1 Click the Issue button for the change to take effect Click Named Variables to produce the display in Figure 3 7 Change variables here as needed Rev 1 0 Copyright O 2012 MED Associates Inc MED ASSOCIATES INC SOF 700RA 29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE Figure 3 6 Displaying Array Displaying Array A from Box 1 A 2 A 3 A 4 Named Vars 1000 3 000 3 000 3 Al A 8 A 3 5 Vars 30 000 180 000 0 100 s 12 13 A 14 0 000 0 000 0 000 Befresh Issue Close Display Data from Box Additional Boxes to Update 0 from Box 1 2 5 1 2 5 000 Select All Deselect Al Figure 3 7 Displaying Named Variables Displaying Named Variables from Box 1 Acclimation Period min METH SIS Number of Trials to Run 30 000 Start Safe Side 1 Left 2 Right 1 000 Vars Stimuli on Safe Side 0 1 Green 2 Yellow 3 3 000 Beneath Stimuli on Shock Side O None 1 Green 2 Yellow 3 000 Stimuli Flash Rate sec 0 500 Delay to Shock Onset on Dark Side sec 0 000 558 Max Shock Duration sec 30 000 No Response Limited Hold Time sec 180 000 Display Data from Additional Boxes to Update Oi 1 we Select All Deselect Al Rev 1 0 Copyright 2012 MED Associates Inc CHAPTER 4 Saving Data
7. Inc MED ASSOCIATES INC SOF 700RA 29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE Figure 3 3 Default Protocol Parameters Acclimation Period min 5 000 Number of Trials to Run 30 000 Start Safe Side 1 Left 2 Right 1 000 Stimuli on Safe Side 0 1 Green 2 Yellow 3 Re 3 000 Stimuli on Shock Side 0 1 Green 2 Yellow 3 000 Stimuli Flash Rate sec 0 500 Delay to Shock Onset on Dark Side sec 0 000 Shock Duration sec 30 000 No Response Limited Hold Time sec 180 000 Shock On Time sec 0 100 The Send Start Command Screen appears next The options available on the screen vary depending upon how many boxes are described in the Hardware Configuration You will be presented with as many boxes as are described in the Hardware Configuration If more than 1 box is in the Hardware Configuration then an option to load more boxes is presented Next the researcher decides to either load more boxes send a start signal to boxes that are already loaded or enter the MED PC IV run time environment without sending a start signal by selecting am finished with the wizard The Run Time screen shown in Figure 3 4 is then displayed Figure 3 4 MED PC Runtime Screen BB MED PC iv File Configure View Macros Help Alo el AX 2 El Sumay Show Rows Al Subject Experi Group Load Program 15 53 Zebrafish Shuttle Closed
8. SOF 700RA 29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE MEDSTATE NOTATION PROCEDURES DOC 264 SOF 700RA 29 USER S MANUAL Rev 1 0 Copyright O 2012 Rights Reserved MED Associates Inc P O Box 319 St Albans Vermont 05478 www med associates com MED ASSOCIATES INC SOF 700RA 29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE Table of Contents HQ 1 Introducti one 1 Chapter 2 dn urhe n o Pul De Dl PK I d 2 Installing Drivers and Software 2 Translating and icr rrr rer rere etre 2 Backing e Pe en oa teet ret 2 Chapter 3 Using the Softwares cometen 3 Using the MED PC IV Load Wizard 5 Viewing Changing Variable 7 Chapter 4 oa ero 9 Saving Data amp Understanding the Raw Data 9 11 Data Analysis Using MED PC to 11 Using a Pre Formatted Table Profile MTP 11 P pl 4 fee T 12 Contact Informmatlor 12
9. User s Manual or contact support med associates com for assistance Figure 2 1 Trans IV Control Panel for Translating and Compiling MedState Notation Code fg Specify Files to Translate Zebrafish Shuttle Box Passive Avoidance Training mpc Make Build No Trans Exclude Cancel Help Backing Up the Software Med Associates strongly encourages creating backup copies of the Zebrafish Shuttle Box programs in case of disk failure When making modifications to the protocol it is advised to keep original copies Rev 1 0 Copyright 2012 MED Associates Inc CHAPTER 3 Using the Software In Zebrafish Shuttle Box Passive Avoidance subjects Zebrafish learn to avoid the environment where an aversive stimulus was previously delivered This test is classically used to test short term or long term memory capabilities In this test an aversive stimulus shock is administered to a Zebrafish once the Zebrafish enters a certain side of the shuttle box This side is typically darkened with a shade or cover Latency to swim to the side of the shuttle box associated with the aversive stimulus is measured for each subsequent trial and is taken as evidence of memory retention The Zebrafish Shuttle Box Passive Avoidance includes both acclimation and testing phases The Zebrafish Shuttle Box has 8 columns of 6 I R photo beam sensors per column illustrated in Figure 3 1 and separate green yellow and red Stimulus Lights o
10. e shock pulse 0 1 seconds e Shock Off Time duration of time between shock pulses 2 4 seconds e Session Time 60 minutes Default values may be modified using the MED PC Experiment Loading Wizard within the protocol or with the Change Variables option Changes made within the protocol Medstate Notation require the code to be translated and compiled to be applied at runtime Notes e Start Side Safe Side e With Zebrafish the shock must be pulsed on and off so that the fish do not become accustomed to the current running through the water Use the Shock On Time and Shock Off Time variables to set the desired timing for the shock pulsation e The program can take measurements as fast as every 10ms for up to eight chambers Rev 1 0 Copyright O 2012 MED Associates Inc MED ASSOCIATES INC SOF 700RA 29 ZEBRAFISH SHUTTLE BOX PASSIVE AVOIDANCE Using the MED PC IV Load Wizard Open MED PC IV and the MED PC Experiment Loading Wizard Welcome screen Figure 3 2 is displayed To proceed with the wizard click Next and the Box Selection screen will be displayed Figure 3 2 Med PC Wizard Welcome Screen To avoid this load wizard deselect the checkbox labeled Run this experiment automatically when starting MED PC Close this screen by clicking the Close button Closing this screen immediately reveals the MED PC Run Time Screen shown in Figure 3 4 To continue using the Wizard press Next On the Box Selection
11. me runs out then the latency to cross will be recorded as the no response time default 180 seconds and the next trial will start immediately The experiment will end when the Max Number of Trials is reached or the Max Session Time is reached Below are the variables used with description default value units and options listed e Duration of Acclimation Period 5 minutes e Number of Trials to Run 30 trials e Start Safe side where the Zebrafish should be when the trial begins the side opposite the Shock side 1 lt 1 left side 2 right side e Stimuli on Safe side setting will correspond to a light that will remain on flash in the Safe side for the duration of the session 3 lt 1 Green 2 Yellow 3 Red gt e Stimuli on Shock side setting will correspond to a light that will remain on flash in the Shock side for the duration of the session 3 lt 1 Green 2 Yellow 3 Red gt e Stimuli Flash Rate the rate each Stimuli light will flash on and off 0 5 seconds e Delay to Shock Onset on Dark Side how much time will elapse between the Zebrafish entrance to the Shock Side and the shock 0 seconds e Max Shock Duration maximum time the shock can last if the Zebrafish fails to escape the Shock Side during the shock 30 seconds No Response Limited Hold Time the longest allowed duration the Zebrafish can spend on the Safe side before a new trial begins 180 seconds e Shock On Time duration of th
12. n the left and right Figure 3 1 IR Beam Numbering as Seen From Front 1 2 3 4 5 6 7 8 The protocol by default starts when the animal is placed in the left side of the chamber When the animal is detected on the start side a 5 minute acclimation period begins The procedure then runs a fixed number of 30 trials A trial proceeds as follows When the Zebrafish swims into the shock zone it will be shocked after the shock delay period The default shock delay is O seconds immediately after crossing into the shock zone The shock is turned off when the Zebrafish escapes from the shock zone or the maximum shock time is reached If the Zebrafish escapes from the shock zone then the next trial starts immediately If the Zebrafish fails to escape from the shock zone before the maximum shock duration runs out then the latency to escape will be recorded as maximum shock duration default 30 seconds the shock will be turned off and the program will turn the Alert signal on Rev 1 0 Copyright O 2012 MED Associates Inc IMPORTANT If the Zebrafhis fails to escape it must be moved to the Start Safe side When the Zebrafish is back on the Start Safe side a START command must be reissued in order for the experiment to continue If the Zebrafish fails to cross into the shock zone before the No Response Limited Hold ti
13. screen the researcher chooses which boxes will be used in the experiment Select the boxes that will run the experiment by clicking in the radio button next to the box number and click Next to proceed The next screen is the Select a Procedure screen This is where the application to be run is selected The screen displays a list of all the currently compiled procedures Select Zebrafish Shuttle Box Passive Avoidance and then click Next The Enter Experiment Data screen will appear next The purpose of this screen is to allow annotations to be added to the data file that is produced by MED PC IV These annotations will help identify the Subject Experiment and Experiment Group upon which data was collected Comments can be added here as well and the data file can be given a customized file name Enter the information desired and click Next the information on this panel is optional and can be skipped if so desired The next screen to appear is the Review Choices screen This is a method of confirming that the information entered is correct If it is not correct select Previous and edit the data If it is correct select Next The Alter Session Parameters Screen is the next screen to appear and displays the default parameter settings as shown in Figure 3 3 The Alter Session Parameters screen allows the researcher to alter the parameters by which a procedure executes Click Next to proceed 5 Rev 1 0 Copyright 2012 MED Associates
14. tes com with a detailed description of the problem or your desired goals so that we may provide you with concise and detailed information Rev 1 0 Copyright O 2012 MED Associates Inc CHAPTER 2 Installing Drivers and Software Please refer to the ENV 012Z X1 Fish Shuttle Box Wiring Guide for hardware setup Before using the software the necessary drivers and software must be installed Install the MED PC software using the instructions in DOC 010 MED PC IV User s Manual Drivers for the computer interface are included on the MED PC installation CD The Zebrafish Shuttle Box Avoidance CD contains the Zebrafish Shuttle Box Passive Avoidance Training mpc MED PC protocol To install the protocol insert the installation CD into the CD ROM player on the PC Click on Install the Zebrafish Shuttle Box software When prompted confirm that MED PC is installed in C MED PC IVA and the installation program will automatically copy the Zebrafish Shuttle Box Passive Avoidance Training mpc to the C MED PC IV MPC folder Translating and Compiling Programs written in MedState Notation need to be translated using Trans IV before they can be executed in this application Open Trans IV and select Translation Translate and Compile Select the Zebrafish Shuttle Box Passive Avoidance Training mpc and click Make Click OK to start the translator If any problems are encountered during this process refer to the on screen help menu DOC 010 MED PC IV
15. xcel File Help Transfer Data Edit Row Profiles Edit Table Profiles Row Transfer Profile NO PROFILE ASSIGNED Select Transfer Orientation Column Labels Horizontal Rows Data C Vertical Columns Table Transfer Profile CAMED PC IM MPC Zebrafish Shuttle Box Passive Avoidance Training Transfer E v Labels Transfer Data Note that the datapath for the Zebrafish Shuttle Box Passive Avoidance Training mtp is listed in the Table Transfer Profile section Select Labels and Data to print data labels and import data Then click Transfer Specify the raw data file to transfer and then click Open The data will transfer to Microsoft Excel spreadsheet Refer to DOC 036 MPC2XL User s Manual for more information on transferring MED PC data files to Microsoft Excel Appendix A Contact Information Please contact MED Associates Inc for information regarding any of our products Visit our website at www med associates com for contact information For technical questions email support med associates com 12 Rev 1 0 Copyright O 2012 MED Associates Inc
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