To design a Standard Operating Procedure that describes the procedure for operation, software, and troubleshooting non-invasive Blood Pressure Systems for small animals.
2.0 SCOPE
This Standard Operating Procedure (SOP) is applicable for the operation, monitoring troubleshooting of non-invasive Blood Pressure Systems for studies conducted at Animal house facilities (AHF)
3.0 RESPONSIBILITY
3.1 All personnel in the Pharmacology Department shall be responsible for implementation and compliance with the SOP.
3.2 Head/Designees/Study Directors shall be responsible for ensuring the implementation and compliance to the SOP.
4.0 DISTRIBUTION
4.1 The Quality Assurance department is responsible for keeping SOP ‘Master Copy’ approved through the Quality Manager.
5.0 DEFINITION(S) & ABBREVIATION(S)
5.1 Definition(s)
5.1.1 NIBP Amplifiers with built-in pumps automatically inflate the tail cuff to occlude the vessel in the tail of a rat or similar small animal, and then slowly deflate the cuff when the inflation point is reached, providing a linear pressure drop. A single control starts both the inflation and deflation cycles, making the system very operator-friendly. Amplifiers have two analog outputs for pressure and pulse waveforms, plus gain adjustment to amplify or attenuate the pulse signal. Systolic, diastolic, and mean BP values.
5.2 Abbreviation(s)
5.2.1 SOP: Standard Operating Procedure
5.2.2 AHF: Animal House Facility
5.2.3 NIBP: Non-invasive Blood Pressure
5.2.4 MRI: Magnetic Resonance Imaging
5.2.5 mmHg: Millimetre of Mercury
6.0 PROCEDURE
6.1 Equipment description
6.1.1 Amplifier order NIBP200A
6.1.2 One tai cuff sensor
6.1.3 One small animal restrainer
6.1.4 MRI-conditional sensor (Optional)
6.1.5 Analog output: pressure 0-3 V DC, Pulse 0-4 V DC
6.1.6 Output cables: Pressure cable and pulse cable
6.1.7 Interface cables: to BIOPAC or third-party A/D hardware
6.1.8 User’s manual
6.2 Specifications
|
Cut-off pressure range |
: |
100 – 300 mmHg (adjustable by 1mmHG steps) |
|
Pressure Accuracy |
: |
300 mmHg Full scale 1% |
|
Pressure Sensitivity |
: |
0.1 mmHg |
|
Pressure Signal output |
: |
300 mmHg/3 Volt DC |
|
Pulse gain levels |
: |
X1, x2, x4, x5, x8, x32 (adjustable) |
|
Pulse display |
: |
Pulse intensity is displayed on A2, derived from plethysmographic
measure The tail sensor detects blood flow and pulse intensity is increased
or decreased, depending on the flow ratio. |
|
Power supply |
: |
12 Volt 2 Amp - External |
|
|
|
|
6.3.1 Connect the CBL150-PRE cable (or CBL35-PRE cable for MP36/35 hardware).
6.3.1.1 BNC to the PRESSURE output on the back panel of the unit.
6.3.1.2 Other end to A1 on the front of the HLT100C/UIM100C (or CH 1 of the MP36/35 unit).
6.3.2 Connect the CBL150-PLS cable (or CBL35-PLS for MP36/35 hardware).
6.3.2.1 BNC to the PULSE output on the back panel of the unit.
6.3.2.2 Other end to A2 on the front of the HLT100C/UIM100C unit (or CH 2 of the MP36/35 unit).
6.3.3 Connect the IRSENSOR.
6.3.3.1 Black cord to the sensor input on the front panel of the NIBP200A.
6.3.3.2 Tubing in the cuff on the front panel of the NIBP200A.
6.3.4 Connect the power.
6.3.4.1 AC300 adapter to the 12 V DC input on the back panel of the NIBP200A.
6.3.4.2 AC300 to Mains power.
6.3.5 Switch the POWER on.
Figure 1: NIBP 200A Front Panel Figure 2:
NIBP 200A Rear Panel
6.4 Animal preparation
6.4.1 Turn the animal chamber on.
6.4.2 Set the temperature value (press and hold P.Set and then press the up or down arrow to reach the desired value).
6.4.2.1 For accurate noninvasive blood pressure measurement, the animal or its tail should be warmed to 32° C.
6.4.3 Press the Heater button to start heating to the selected temperature value.
6.4.4 Place the animal inside the RESTRAINER “Animal Holder” (select the suitable size for the animal volume).
6.4.4.1 Leave the tail outside.
6.4.4.2 Adjust the length to obtain a position where the animal has limited movement.
6.4.5 Place the RESTRAINER (with the animal) in the heating section of the Animal Heating Chamber.
6.4.6 Wait approximately 30 minutes for the animal to reach the selected temperature.
6.4.7 Remove the RESTRAINER from the Animal Heating Chamber.
6.4.8 Connect the IRSENSOR to the tail of the animal inside the RESTRAINER.
6.4.9 Check if the sensor just fits to the tail. The sensor should be between the midpoint of tail and tail end (spinal column). To achieve this, a suitable sensor should be selected.
6.4.10 Wait for the animal to relax and become inactive before starting measurements.
Figure 3:
Restrainer animal holder
Figure 4: Tail cuff/sensor
Figure 5: Optional heating chamber
6.5 Software setup
6.5.1 Launch AcqKnowledge 4.x.
6.5.2 Select the “Create/Record a new experiment” option.
6.5.3 Select “MP160/150 > Set Up Data Acquisition > Channels > “Add New Module...”
6.5.3.1 From the new module list, select HLT100C-A1 (MP160) or UIM100C-A1(MP150), (or whichever channel the CBL150-PRE pressure cable is connected to) and click “Add.”
6.5.3.2 From the HLT100C (MP160) or UIM100C (MP150) Transducer list, select “NIBP200A – Small Animal Tail BP, Pressure” or “NIBP250 – Small Animal Tail BP, Pressure” and click OK.
6.5.3.3 Click “Calibrate” in the resulting Calibration dialog.
6.5.4.1 From the new module list, select HLT100C-A2 (MP160) or UIM100C-A2 (MP150) (or whichever channel CBL150-PLS pulse cable is connected to) and click “Add.
6.5.4.2 From the HLT100C (MP160) or UIM100C (MP150) Transducer list, select “NIBP200A – Small Animal Tail BP, Pulse” or “NIBP250 – Small Animal Tail, Pulse” and click OK.
6.5.5 Launch the BIOPAC software.
6.5.6 Choose “MP menu > Set up Channels.”
6.5.7.1 If desired, enter channel Labels: A1 Pressure and A2 Pulse.
6.5.8 Calibrate for the pressure measurement of IRSENSOR.
6.5.8.1 Select A1 (Pressure) click Setup and establish these settings:
6.5.10 Calibrate for the pulse measurement of IRSENSOR.
6.5.10.1 Ensure that the tail is not inside the IRSENSOR it is empty, and the sensor resides freely.
6.5.10.2 Select A2 (Pulse) click Setup and establish these settings:
6.5.11 Choose “MP menu > Set up Acquisition” and establish the following settings:
6.5.11.1 Mode = Record and Append to Memory
6.5.11.2 Sample Rate = 200 samples/second
6.5.11.3 Total Length = 24 seconds
6.5.11.4 Repeat = every 3 seconds for 10 times
6.5.13 Choose “MP menu > Setup Trigger” and establish the following settings:
6.5.13.1 Trigger = CH 1, Pos Edge
6.5.13.2 Trigger Level = 0.33 Volts
6.5.13.3 (based on 1 V ≈ 100 mmHg)
6.5.13.4 Delay = 0 samples
6.5.15 Software setup for acknowledge 4.x or BSL 4.x with MP3x Hardware
6.5.15.1 Launch the software.
6.5.15.2 Select the “Create/Record a new experiment” option.
6.5.15.3 If necessary, choose “MP3x > Set up Data Acquisition > Channels.”
6.5.15.4 Enable analog inputs CH1 and CH2 and select the Acquire, Plot, and Value options.
6.5.15.6 Click “New Channel Preset,” enter “NIBP200A-Pressure” and click OK.
6.5.15.7 Establish the following settings:
6.5.15.7.1 Channel Preset = NIBP200A-Pressure
6.5.15.7.2 Channel Label = CUFF PRESSURE
6.5.15.7.3 Gain = x10
6.5.15.7.4 Input Coupling = DC
6.5.15.7.5 Filter = 1
6.5.15.7.6 Type = Low Pass
6.5.15.7.7 Frequency= 30
6.5.15.7.8 Q = 0.5
6.5.15.8.1 Click the “Scaling” button and establish the following settings:
6.5.15.8.2 Map values
6.5.15.8.3 Cal1 = 0
6.5.15.8.4 Cal2 = 100
6.5.15.8.5 Units label = mmHg
6.5.15.8.6 Click the Cal 1 button.
6.5.15.8.7 Add “333” to the Cal 1 Input value, and enter the result in
6.5.15.8.8 Cal 2 Input value (Cal 2 = Cal 1 + 333)
6.5.15.8.9 Click OK as needed to exit the CH1 “Scaling” and Input “Channel” setup dialogs.
6.5.15.10 Click “New Channel Preset,” enter “NIBP200A-Tail Pulse” and click OK.
6.5.15.11 Establish the following settings:
6.5.15.11.1Channel Preset = NIBP200A-Tail Pulse
6.5.15.11.2Channel Label = TAIL PULSE
6.5.15.11.3Gain = x10
6.5.15.11.4Input Coupling = DC
6.5.15.11.5Filter = 1
6.5.15.11.6Type = Low Pass
6.5.15.11.7Frequency = 50
6.5.15.11.8Q = 0.5
6.5.15.12.1 Ensure that the tail is not inside the IR SENSOR and that the sensor resides freely.
6.5.15.12.2Click “Scaling” button and establish the following settings:
6.5.15.12.3Map values
6.5.15.12.4Cal 1 = 0
6.5.15.12.5Cal 2 = 1000
6.5.15.12.6Units label = mV
6.5.15.12.7 Click the Cal 1 button.
6.5.15.12.8 Add “333” to the Cal 1 Input value and enter the result in the Cal 2 Input value (Cal 2 = Cal 1 + 333).
6.5.15.12.9 Click OK as needed to exit the CH2 “Scaling” and “Input Channel” setup dialogs.
Mode = Record and Append using Memory
Sample Rate = 200 samples/second
Acquisition Length = 24 seconds
Repeat = every 3 seconds for 10 times
6.5.15.14 Choose “Trigger” and establish the following settings.
Trigger = CH 1, Pos Edge
Trigger Level = 30 mmHg
6.5.15.15 Exit the Data Acquisition
Settings dialog.
6.5.16.1 Confirm that the animal is ready and that the IRSENSOR is attached to the tail.
6.5.16.2 Click “Start” in the BIOPAC software window.
6.5.16.3 Press the START button on the front panel of NIBP200A.
6.5.16.4 IRSENSOR will pump up the Cuff automatically.
6.5.16.5 When the Cuff Pressure on A1 reaches 30 mmHg. the cuff pressure and tail pulse signals will be generated.
6.5.16.6 The recording will stop automatically after 24 seconds.
6.5.16.7 Press START to continue with the next measurement and repeat as necessary.
6.5.16.8 Choose File > Save or Save as when done.
6.5.16.2 Click “Start” in the BIOPAC software window.
6.5.16.3 Press the START button on the front panel of NIBP200A.
6.5.16.4 IRSENSOR will pump up the Cuff automatically.
6.5.16.5 When the Cuff Pressure on A1 reaches 30 mmHg. the cuff pressure and tail pulse signals will be generated.
6.5.16.6 The recording will stop automatically after 24 seconds.
6.5.16.7 Press START to continue with the next measurement and repeat as necessary.
6.5.16.8 Choose File > Save or Save as when done.
6.5.17
NIBP200a analysis
Calculation of Systolic, Diastolic, and Mean
6.5.17.2 Select from the list Max, Min, and Mean for three different Labels.
6.5.17.3 Select Channel 1 as a channel option.
6.5.17.4 Select cursor ‘I’ from the cursor option on the bottom right of the screen.
6.5.17.5 On the graphical display, starting from the point of the first pulse, select an area to the maximum.
6.5.17.6 Review the results for Max (Systolic), Min (Diastolic), and Mean measurements.
6.5.17.8 Use the I-beam cursor to select the maximum points of the peaks of the CH2 pulse waveform.
6.5.17.9 Review the results for BPM (Heart Rate value) for each peak.
6.6 Troubleshooting
6.6.1 Tail pulse signals are not regular
6.6.1.1 The animal may be under stress, resulting in excessive tail movement. Remove the animal from the restrainer holder until it calms down before continuing with the experiment.
6.6.1.2 The tail may not be sufficiently warmed or cooled down. Put the animal in the Tail Heater Chamber and repeat the heating process. Make sure the tail temperature is 32° C.
6.6.1.3 Tail Cuff sizing may be incorrect. Check Table 5 on the following page for sizing descriptions.
6.6.1.4 Tail Cuff Sensor position may be incorrect. Try re-attaching the sensor in a different location on the tail. The optimal location is between the mid-point of the tail and the base of the tail (spinal column).
6.6.2 Compressor is working continuously
6.6.2.1 Immediately turn off the NIBP system.
6.6.2.2 Remove the Tubing from the Cuff connector on the panel of the NIBP system.
6.6.2.3 Turn the system back on.
6.6.2.4 Close the air outlet by pressing the finger on the Cuff output and press the “Start” button. The compressor will work for a few seconds and stop (please inform BIOPAC if the Compressor does not stop). The pressure chart should be viewable on the screen.
6.6.2.5 If the Compressor stops automatically, it means that the system is working normally.
6.6.3 There is leakage in the tubing connections and Cuff of the IRSENSOR
6.6.3.1 Make sure the tubing is securely attached.
7.0 PRECAUTIONS
7.1 General laboratory procedures should be followed, which include: no eating, no chewing gum, no drinking, and no applying cosmetics in the work area. Laboratory coats and gloves must be worn at all times in the work area unless the protocol specifically describes the appropriate attire for the procedure.
8.1 Reference
8.1.1 Refer to the BIOPAC Multi-Channel, Computerized, Non-Invasive Blood Pressure System for Mice and Rats operation manual for additional information.
Format – I Format for Log Book Maintenance
Format- II Format for Document History Sheet
9.0 REVISION HISTORY
9.1.1 Maintain a record of all SOP revisions, including the version number, date, description of changes, and the names of the individuals who prepared, reviewed, and approved the revision. This record should be kept along with the current version of the SOP.
END OF THE DOCUMENT
You may like to read these links:
1. List of All SOPs and Documents for the Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for the In-vivo Laboratory.
4. List of All SOPs and Documents for Clinical Research
5. List of Instruments and Equipment Used In Laboratory Animal Facility
6. Animal Facility Design - Small Laboratory Animals (Rat, Mice, Rabbit, Guinea Pig)
7. List of Chemicals Needed for In-vivo Laboratory
8. List of Physiological Data for Small Laboratory Animals
9. List of All SOPs and Documents for the Animal House Facility
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