1.0 PURPOSE
To design a Standard Operating Procedure that describes the procedure for operation, maintenance of Tail flick unit.
2.0 SCOPE
This Standard Operating Procedure (SOP) is applicable for the operation, maintenance of Electronic Von Frey for Pharmacology studies .
3.0 RESPONSIBILITY
3.1 All personnel in the Pharmacology Department shall be responsible for implementation and compliance to the SOP.
3.2 Head/Designees/Scientists/Study Directors of Pharmacology department shall be responsible to ensure the implementation and compliance to the SOP.
4.0 DISTRIBUTION
4.1 The quality assurance department responsible to keep SOP ‘Master Copy’ approved through Quality Assurance Head.
4.2 The copy of ‘Control Copy’ of all SOPs is being distributed in Pharmacology department and placed near related Equipment/ Instrument as ‘Display Copy’.
5.0 DEFINITION(S)
5.1 Definition(s)
5.1.1 The instrument measures the latency of the avoidance response when pain is induced by radiant heat applied to the animal’s tail.
5.2 Abbreviation(s)
5.2.1 SOP : Standard Operating Procedure
5.2.3 USB : Universal Serial Bus
5.2.4 IR : Infrared
6.0 PRECEDURE
6.1 Principle
6.1.1 The rat is held by the operator on the instrument unobstructed upper panel (see 1, figure1) in such a way that its tail, placed over a flush mounted window, receives the I.R. energy.
6.1.2 The operator starts the stimulus and the related reaction-time counter by the pedal switch or by function key located on the front panel.
6.1.3 When the rat feels pain and flicks its tail, a sensor detects it, stops the reaction-time counter and switches off the bulb.
6.1.4 The reaction time of the animal is thus automatically determined to the nearest 0.1 second.
Figure-1: IR Instrument Setup
Figure-2: IR Instrument Setup
6.2 Instrument description
6.2.1 It basically consists of an I.R. source (50W; bulb), whose radiant energy of adjustable intensity is focused by an embodied parabolic mirror on the rat tail.
6.2.2 The instrument components are neatly arranged in a box of new design, which contains the I.R. source, the sensor, the microcontroller and the electronic circuit.
6.2.3 The multifunction liquid-crystal graphic-display monitors the withdrawal latency to the nearest 0.1 second; latency time is the time elapsing from the moment the START key is activated to the moment the animal flicks its tail.
6.2.4 The graphic display presents all available command: the operator sets the experiment configuration via the command keyboard located on the right of the display.
6.3 Graphic Display
6.3.1 When the instrument is switched on, our logo with the two mice is displayed. By depressing F4 (ESC), you enter the user area.
6.3.2
The four
lines of the top area show:
7.3.2.1
The actual date
7.3.2.2
The actual time
7.3.2.3
The instrument serial number
7.3.2.4
The firmware version
6.3.3
The right
portion of the display window shows the abbreviation of the key function,
meaning:
7.3.2.5
OPR Operation
7.3.2.6
FNC Function
7.3.2.7
MEM Memory
7.3.2.8
ESC Escape
6.3.4
The F4
key, which has the function of ESCAPE key, enables the operator to leave a menu
and go back to a previously selected one.
Table
1: Key abbreviations
|
Abbreviations |
Explanation |
|
CAL |
Calibration |
|
CFG |
Configuration |
|
CLK |
Clock setting |
|
CUT |
Cut-off time |
|
DIS |
Display contrast |
|
DWN |
Decrement |
|
ENT |
Enter |
|
EPR |
External printer |
|
ESC |
Escape |
|
FNC |
Function menu |
|
FTR |
Feature menu |
|
IR |
I.R. intensity |
|
MEM |
Memory menu |
|
MPR |
MiniPrinter |
|
MSK |
Memory stick |
|
NO |
No |
|
NUL |
Annulment |
|
OK |
Confirm |
|
OPR |
Operation menu |
|
OUT |
Out |
|
PRN |
Data printing selection |
|
RES |
Data memory reset |
|
STR |
Start |
6.4 Installation
6.4.1 Notes on the Instruction Manual
7.4.1.1 The Tail Flick Instruction Manual and SOP shall be thoroughly read before installation and operation of the instrument.
6.4.2 Assembling the Equipment
7.4.1.2 For the first time use, Tail Flick unit shall be positioned on a stable and flat bench or table surface.
7.4.1.3 Fit the top panel extension, as indicated in figure below, and secure it by tightening the two screws.
6.4.3 Before Applying Power
7.4.1.4 Consider the Power Module on the back panel of the Tail Flick, which includes (from left to right):
7.4.1.4.1 The inlet connector of the
7.4.1.4.2 Mains cord
7.4.1.4.3 The mains switch
7.4.1.4.4 The fuse holder
Figure 3: Power Module
6.5 Operation
6.5.1 Switching On
7.5.1.1 Switch on the instruments; the software takes some seconds to perform a complete checking of the electronic circuit.
7.5.1.2 During the checking, the display shows the mice logo. By depressing ESC, the display shows the software version which is actually installed.
7.5.1.3 The software version is necessary in case of communication with our laboratory for operational or servicing problems.
6.5.2 Display Contrast
7.5.1.4 To modify the display contrast, depress the F2 (FNC) key from the Main Menu, then the F3 (DIS) key. The display shows:
7.5.1.5 Keep the arrow keys F1 (UP) or F2 (DWN) depressed until you reach the desired display contrast.
7.5.1.6 The readability of the liquid crystal display depends on the angle of view: select the ideal contrast according to the height of the table, the operator’s distance and so on.
6.5.3 Setting IR intensity
6.5.4 Before carrying out the test on the animal, set the I.R. intensity. Enter the I.R. setting menu, by depressing F1 (OPR), F2 (SET) and F1 (IR ). The following display will appear:
6.5.5 Now use the F1 (UP) or F2 (DWN) keys to set intensity to the desired value.
6.5.6 How to Set a Customized Experimental Configuration
7.5.1.7 This function enables the operator to personalize the basic datum by adding, in sequence:
7.4.1.4.5 Sequence number
7.4.1.4.6 Experiment number
7.4.1.4.7 Animal gender
6.5.7 From the main menu, depress the F2 (FNC) key, followed by the F1 (OUT) key. Depress again the key F2 (FTR); the display shows the first setting, the sequence number.
6.5.8 All the available selections for each additional information are displayed in a loop. Scan them by the F1 (UP) or F2 (DWN) keys, select the suitable one and confirm the selection by depressing the F4 (OK) key, or avoid one or more features by depressing the F3 (NUL) key.
6.5.9 The selected features will be maintained even if the instrument is turned off. To erase one or more features previously set, simply enter this software level, highlight the concerned feature and depress the F3 (NUL) key.
6.5.10 After entering the last feature (M or F), the software goes automatically back to the main menu.
6.5.11 Sequence number
6.5.12 The first additional information to be defined is the sequence number. This section is highlighted as soon as the F2 (FTR) key is depressed.
6.5.13 Use the F1 (UP) or F2 (DWN) key to set the desired sequence number from 01 to 99 and confirm by depressing F4 (OK).
6.5.14 The sequence number will automatically increase by one figure at any subsequent measurement. It trips once a datum has been acquired and recorded, indicating the fresh sequence number ready for the next measurement.
6.5.15 If a sequence number is not necessary, depress the F3 (NUL) key.
6.5.16 Experiment Number
7.5.1.8 As soon the F4 (OK) or F3 (NUL) keys are depressed, followed by the F4 key again, the second option is highlighted.
7.5.1.9 The experiment number, from 1 to 9, can now be selected via the F1 (UP) or F2 (DWN) key and confirmed via the F4 (OK) key.
6.5.17 Animal Gender
7.5.1.10 After the experiment number, it is possible to define the gender of the animal to be processed: MALE/FEMALE.
7.5.1.11 Select it by depressing the F1 (U) or F2 (DWN) key and confirm it via the F4 (OK) key; avoid the gender selection by depressing the F3 (NUL) key.
6.5.18 Cut-Off Time
7.5.1.12 To get over a “false start”, i.e., in case the beam, misaligned with the aimed tail area, has been accidentally triggered, and to avoid useless overheating of the tail when the animal does not react within the expected reaction time, a cut-off time can be set.
7.5.1.13 The present value indicates the time (in seconds) after which the experiment will be interrupted.
7.5.1.14 Select the cut-off time by depressing theF1 (OPR), F2 (SET) and F2 (CUT).
7.5.1.15 Depress the F1 (UP) or F2 (DWN) key until you reach the desired value and confirm it via the F4 (OK) key.
6.5.19 Configuration File
7.5.1.16 Up to 6 different experiments can be configured, following the instructions outlined in previous paragraphs, saved in the memory key, ready to be recalled to the PC and/or to the instrument.
7.5.1.17 The involved parameters are:
7.4.1.4.8 I.R. Intensity
7.4.1.4.9 Cut-Off Time
7.4.1.4.10 Experiment Number
7.4.1.4.11 Gender
7.4.1.4.12 Sequence Number
7.4.1.4.13 MiniPrinter
6.6 Calibration
6.6.1 The calibration consists of the measurement of the I.R. output power of the Tail Flick with an I.R. Radiometer1, which in turn has been calibrated with a standard I.R. source.
6.6.2 To enter the calibration menu, depress F1 (OPR), followed by the F3 (CAL) key.
6.6.3
Radiometer
7.6.3.1
The
Radiometer enables us to deliver, and the user to maintain, the I.R. source of
each.
7.6.3.2
Tail
Flick Unit with identical power flux output (mW per square cm).
7.6.3.3
There is
no way to calibrate the Tail Flick Unit (as any other I.R. emitting device)
without a Radiometer; the researcher should consider this extremely useful
accessory, the optional I.R. Radiometer Cat. 37300, a battery operated self-sufficient
instrument, complete with I.R. probe, digital meter and adaptors for the Tail
Flick (and Plantar Test), all parts neatly lodged in a sturdy plastic case.
6.6.4
Calibration without I.R. Radiometer
7.6.4.1
Calibrating
the Tail Flick Unit without the I.R. Radiometer, means to recall the default
calibration value set at our factory during the testing phase.
7.6.4.2
From the
Calibration Menu, depress the F1 (DEF) key and proceed by the F3 (YES) key, to
confirm the change. The default calibration value is thus recalled.
6.7 Maintenance
6.7.1 Unplug the main cord before carrying out any maintenance job.
6.7.2 To inspect and/or replace the fuses, disconnect the mains cable first! Insert a miniature Screw driver in the slot indentation and snap out the slide which houses the fuses.
6.7.3 Tail Flick Unit universal mains input accepts any voltage from 85 to 264 Volt AC., for which we recommend the use of 2A fast-blow fuses (type F2A).
6.7.4 Battery replacement
7.7.4.1 Tail Flick Unit memory is supported by a battery snaphat type M4T32-BRI2SH1 of ST. The rated battery endurance is 10 years, therefore replacing the batteries is not a frequent task.
7.7.4.2 When the batteries are exhausted, the display shows the caption “MEMORY LOSS!!”
6.7.5 Replacing the bulb
7.7.5.1 You will realize, in the course of an experiment that the bulb is burnt-out (glow lamps have unfortunately a finite life span) because the missing I.R. source causes the counter to stop automatically after the preset cut-off time.
7.7.5.2 But even before that, you will have noticed the absence of any animal reaction! The bulb is located into the I.R. Source.
7.7.5.3 To replace the bulb and, in general, to service the I.R. Source, it is necessary to remove the top and bottom panels.
7.7.5.4 To take the bulb out, first loose the two miniature contact screws which fasten the socket to the bulb current-carrying studs. Swing the socket apart.
7.7.5.5 Free the bulb by removing its holding collar. You will notice the small protrusion on the glass rim of the bulb reflector.
7.7.5.6 Position the fresh bulb into its pentagonal receptor and rotate it until the mentioned protrusion mates the rectangular cut of the supporting winglet. Try to turn the bulb, both CW & CCW. If it doesn’t, it means the fit is O.K. Reposition the collar.
7.7.5.7 Then fit the socket to the studs of the fresh bulb and retighten the contact screws. There is no polarity to worry about. Avoid leaving “finger prints” on the concave reflector, while you are handling the bulb.
7.7.5.8 Needless to add, recalibrate.
6.7.6 Cleaning the optics thoroughly
7.7.6.1 Fan cooling, using room air, and protecting the I.R. optics from dust are both laudable but – alas – conflicting requirements.
7.7.6.2 After 2-3 years and during major instrument overhauls, a thorough cleaning of the optics is advisable: it can be done at the factory only, where appropriate tools and facilities are available for dismantling, cleaning (ultrasonic bath), reassembling, realigning, etc.
7.7.6.3 Please contact our Customer Support.
6.7.7 Long inactivity
7.7.7.1 The instrument does not require any particular maintenance after long inactivity, except cleaning. Calibration is recommended, if the Radiometer 37300 is available.
7.0 PRECAUTIONS
7.1 Do not attempt to open or perform any service work
7.2 Do not connect up human subjects
7.3 Tail Flick instrument shall be placed on a flat surface.
7.4 Do not obstruct free and comfortable access to the power module.
7.5 Use original accessories and spare parts only.
7.6 Immediately disconnect and replace a damaged mains cable.
7.7 Do not operate the instrument in hazardous environments or outside prescribed environmental limitations (i.e. +18C°/+24C°, 60% relative humidity, non-condensing).
7.8 Do not spray any liquid on the connectors.
7.9 Keep inflammables far from the heating source.
7.10 Read and understands the following key abbreviations as shown in table 1 before proceed to work.
8.0 REFERENCES
8.1 References
8.1.1 F.E. D’Amour & D.L. Smith: ”A Method for Determining Loss of Pain Sensation” J. Pharmacol. Exp. Therap. 72: 74-79, 1941.
8.1.2 Ugo Basile. Instruction Manual. Tail-Flick Unit. Cat. No. 37360
END OF THE DOCUMENT
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