Here we present a combination of laser Doppler perfusion imaging (LDPI) and laser Doppler perfusion monitoring (LDPM) to measure spinal cord local blood flows and oxygen saturation (SO2), as well as a standardized procedure for introducing spinal cord trauma on rat.
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Method Article
Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat
* These authors contributed equally
In This Article
Summary
Abstract
Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory alterations of the spinal cord. In this article, our goal was to use both Laser Doppler imaging and monitoring to analyze the change of BF after spinal cord injury. Both the laser Doppler image scanner and the probe/monitor were being employed to obtain each readout. The data of LDPI provided a local distribution of BF, which gave an overview of perfusion around the injury site and made it accessible for comparative analysis of BF among different locations. By intensely measuring the probing area over a period of time, a combined probe was used to simultaneously measure the BF and oxygen saturation of the spinal cord, showing overall spinal cord perfusion and oxygen supply. LDF itself has a few limitations, such as relative flux, sensitivity to movement, and biological zero signal. However, the technology has been applied in clinical and experimental study due to its simple setup and rapid measurement of BF.
Introduction
The tissue of the spinal cord is highly vascularized and extremely sensitive to hypoxia induced by spinal cord injury (SCI). Our previous studies showed that blood flow of the spinal cord was significantly decreased after concussion injury1,2, which might be related to the deficit of motor function. Recent studies have shown that the integrity of blood vessels following SCI is well-correlated with the improvement of sensory motor function3. It has been reported that improved vascularity might rescue white matter, indirectly leading to improved function4. Therefor....
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Protocol
Animal protocols involving experimental animals followed guidelines established by the National Institutes of Health (NIH) and were approved by the Animal Care and Use Committee of Capital Medical University.
The procedures of introducing SCI and measuring BF of spinal cord using laser Doppler equipment described below were used in a published study1.
1. Preparation for the Surgery
- Prepare pentobarbital sodium solution 3% (w/v) in saline and administrate at the dose of 35 mg/kg.
Caution: pentobarbital sodium is a controlled substance. Detailed records should be kept....
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Results
LDPI was used to measure BF in the spinal cord, which was quantified along the rostral-caudal axis of the spinal cord by extracting linear profiles (Figure 4). Figure 5A and Figure 5B represent the flux imaging of the spinal cord of the sham group and SCI group, respectively. Figure 5C and Figure 5D represent the altering BF along the rostra.......
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Discussion
A few details should be noticed when performing this protocol. Firstly, the process of anesthesia and surgery should be carried out as quickly and elegantly as possible to minimize the introduced stress to the animal. To reduce disturbance to the results, keep the animal in a relatively peaceful and stable state. Secondly, more attention should be paid to bleeding during the measurement using laser Doppler equipment, since blood could potentially interfere with the reading. Finally, during the data recording, animals sho.......
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Disclosures
The authors have nothing to disclose.
Acknowledgements
The authors have no acknowledgements.
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Laser Doppler Line Scanner | Moor Instruments | moorLDLS2 | |
| Laser Doppler Monitor | Moor Instruments | moorVMS-LDF | |
| Probe for Monitor | Moor Instruments | VP3 | Blunt needle end delivery probe |
| Impactor | Precision Systems and Instrumentation | IH-0400 | |
| Phenobarbital sodium | Sigma-Aldrich | P3761 | |
| Buprenorphine | Sigma-Aldrich | B-908 | |
| Syringe | Becton Dickinson Medica (s) Pte.Ltd | 300841 | |
| Surgical suture needles with thread | Shanghai Pudong Jinhuan Medical Products Co., Ltd | 18T0329 (batch number) /4-0 | |
| Scalpel | Operation instrument factory of Shanghai Medical Instrument Co., Ltd. | J11030 4# | |
| Scalpel blade | Operation instrument factory of Shanghai Medical Instrument Co., Ltd. | J12130 20# | |
| Ophthalmic forceps | Operation instrument factory of Shanghai Medical Instrument Co., Ltd. | JD1040 | |
| Hemostatic forceps | Operation instrument factory of Shanghai Medical Instrument Co., Ltd. | J31050 | |
| Benzyl penicillin sodium | North China Pharmaceutical Co., Ltd | F6072116 (batch number) | |
| 75% alcohol | Dezhou Anjie Gaoke disinfection products Co., Ltd | 150421R (batch number) | |
| Iodine | Shandong Lierkang Medical Technology Co., Ltd | 20170102 (batch number) | |
| Rat | Laboratory Animal Center, The Academy of Millitery Medical Sciences | Sprague-Dawly (rat strain) |
References
- Jing, Y. L., Bai, F., Chen, H., Dong, H. Meliorating microcirculatory with melatonin in rat model of spinal cord injury using laser Doppler flowmetry. Neuroreport. 27 (17), 1248-1255 (2016).
- Jing, Y. L., Bai, F., Chen, H., Dong, H.
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