Name: using laser doppler imaging and monitoring to analyze spinal cord microcirculation in rat
Uploaded: 2018-05-30T13:00:00
Description: Watch this Scientific Journal Video about Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat at JoVE.com

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Protocolli degli animali che coinvolgono gli animali sperimentali seguivano linee guida stabilite dai National Institutes of Health (NIH) e sono stati approvati dalla cura degli animali e uso Comitato di Capital Medical University.
Le procedure di introduzione di SCI e di misurazione BF del midollo spinale utilizzando laser Doppler apparecchiature descritte di seguito sono state utilizzate in un studio pubblicato1.
1. preparazione per la chirur...

<ol>
	<li>Jing, Y. L., Bai, F., Chen, H., Dong, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Meliorating+microcirculatory+with+melatonin+in+rat+model+of+spinal+cord+injury+using+laser+Doppler+flowmetry.">Meliorating microcirculatory with melatonin in rat model of spinal cord injury using laser Doppler flowmetry.</a> Neuroreport. 27 (17), 1248-1255 (2016).</li><li>Jing, Y. L., Bai, F., Chen, H., Dong, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Melatonin+prevents+blood+vessel+loss+and+neurological+impairment+induced+by+spinal+cord+injury+in+rats.">Melatonin prevents blood vessel loss and neurological impairment induced by spinal cord injury in rats.</a> J Spinal Cord Med. , 1-8 (2016).</li><li>Han, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rescuing+vasculature+with+intravenous+angiopoietin-1+and+alpha+v+beta+3+integrin+peptide+is+protective+after+spinal+cord+injury.">Rescuing vasculature with intravenous angiopoietin-1 and alpha v beta 3 integrin peptide is protective after spinal cord injury.</a> Brain. 133 (Pt 4), 1026-1042 (2010).</li><li>Gerzanich, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=De+novo+expression+of+Trpm4+initiates+secondary+hemorrhage+in+spinal+cord+injury.">De novo expression of Trpm4 initiates secondary hemorrhage in spinal cord injury.</a> Nat Med. 15 (2), 185-191 (2009).</li><li>Phillips, J. P., Cibert-Goton, V., Langford, R. M., Shortland, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perfusion+assessment+in+rat+spinal+cord+tissue+using+photoplethysmography+and+laser+Doppler+flux+measurements.">Perfusion assessment in rat spinal cord tissue using photoplethysmography and laser Doppler flux measurements.</a> Journal of Biomedical Optics. 18 (3), 037005 (2013).</li><li>Garcia-Lopez, P., Martinez-Cruz, A., Guizar-Sahagun, G., Castaneda-Hernandez, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+spinal+cord+injury+changes+the+disposition+of+some,+but+not+all+drugs+given+intravenously.">Acute spinal cord injury changes the disposition of some, but not all drugs given intravenously.</a> Spinal Cord. 45 (9), 603-608 (2007).</li><li>Brown, A., Nabel, A., Oh, W., Etlinger, J. D., Zeman, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perfusion+imaging+of+spinal+cord+contusion:+injury-induced+blockade+and+partial+reversal+by+&#946;2-agonist+treatment+in+rats.">Perfusion imaging of spinal cord contusion: injury-induced blockade and partial reversal by &#946;2-agonist treatment in rats.</a> Journal of Neurosurgery-Spine. 20 (2), 164-171 (2014).</li><li>Olive, J. L., McCully, K. K., Dudley, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+flow+response+in+individuals+with+incomplete+spinal+cord+injuries.">Blood flow response in individuals with incomplete spinal cord injuries.</a> Spinal Cord. 40 (12), 639-645 (2002).</li><li>Yamada, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+blood+flow+and+pathophysiological+changes+after+transient+spinal+cord+ischemia+in+cats.">Spinal cord blood flow and pathophysiological changes after transient spinal cord ischemia in cats.</a> Neurosurgery. 42 (3), 626-634 (1998).</li><li>Gordeeva, A. 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Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-treatment+with+LCZ696,+an+orally+active+angiotensin+receptor+neprilysin+inhibitor,+prevents+ischemic+brain+damage.">Pre-treatment with LCZ696, an orally active angiotensin receptor neprilysin inhibitor, prevents ischemic brain damage.</a> Eur J Pharmacol. 762, 293-298 (2015).</li><li>Vertiz-Hernandez, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=L-arginine+reverses+alterations+in+drug+disposition+induced+by+spinal+cord+injury+by+increasing+hepatic+blood+flow.">L-arginine reverses alterations in drug disposition induced by spinal cord injury by increasing hepatic blood flow.</a> J Neurotrauma. 24 (12), 1855-1862 (2007).</li><li>Garcia-Lopez, P., Martinez-Cruz, A., Guizar-Sahagun, G., Castaneda-Hernandez, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+spinal+cord+injury+changes+the+disposition+of+some,+but+not+all+drugs+given+intravenously.">Acute spinal cord injury changes the disposition of some, but not all drugs given intravenously.</a> Spinal Cord. 45 (9), 603-608 (2007).</li><li>Li, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post+pressure+response+of+skin+blood+flowmotions+in+anesthetized+rats+with+spinal+cord+injury.">Post pressure response of skin blood flowmotions in anesthetized rats with spinal cord injury.</a> Microvasc Res. 78 (1), 20-24 (2009).</li><li>Boyle, N. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scanning+laser+Doppler+is+a+useful+technique+to+assess+foot+cutaneous+perfusion+during+femoral+artery+cannulation.">Scanning laser Doppler is a useful technique to assess foot cutaneous perfusion during femoral artery cannulation.</a> Critical Care. 3 (4), 95-100 (1999).</li><li>Emmanuel, A. V., Chung, E. A. L., Kamm, M. A., Middleton, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+gut-specific+autonomic+testing+and+bowel+dysfunction+in+spinal+cord+injury+patients.">Relationship between gut-specific autonomic testing and bowel dysfunction in spinal cord injury patients.</a> Spinal Cord. 47 (8), 623-627 (2009).</li><li>Sheu, J. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combination+of+cilostazol+and+clopidogrel+attenuates+rat+critical+limb+ischemia.">Combination of cilostazol and clopidogrel attenuates rat critical limb ischemia.</a> J Transl Med. 10, 164 (2012).</li><li>Basso, D. M., Beattie, M. S., Bresnahan, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Graded+histological+and+locomotor+outcomes+after+spinal+cord+contusion+using+the+NYU+weight-drop+device+versus+transection.">Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection.</a> Experimental Neurology. 139 (2), 244-256 (1996).</li><li>Basso, D. M., Beattie, M. S., Bresnahan, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Sensitive+and+Reliable+Locomotor+Rating-Scale+for+Open-Field+Testing+in+Rats.">A Sensitive and Reliable Locomotor Rating-Scale for Open-Field Testing in Rats.</a> Journal of Neurotrauma. 12 (1), 1-21 (1995).</li><li>Oberg, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+motion--a+disturbance+in+the+laser-Doppler+blood+flow+signal?.">Tissue motion--a disturbance in the laser-Doppler blood flow signal?.</a> Technol Health Care. 7 (2-3), 185-192 (1999).</li><li>Tenland, T., Salerud, E. G., Nilsson, G. E., Oberg, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+and+temporal+variations+in+human+skin+blood+flow.">Spatial and temporal variations in human skin blood flow.</a> Int J Microcirc Clin Exp. 2 (2), 81-90 (1983).</li><li>Kernick, D. P., Tooke, J. E., Shore, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biological+zero+signal+in+laser+Doppler+fluximetry+-+origins+and+practical+implications.">The biological zero signal in laser Doppler fluximetry - origins and practical implications.</a> Pflugers Arch. 437 (4), 624-631 (1999).</li><li>Rudolph, A. M., Heymann, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+circulation+of+the+fetus+in+utero.+Methods+for+studying+distribution+of+blood+flow,+cardiac+output+and+organ+blood+flow.">The circulation of the fetus in utero. Methods for studying distribution of blood flow, cardiac output and organ blood flow.</a> Circ Res. 21 (2), 163-184 (1967).</li><li>Dubory, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contrast+Enhanced+Ultrasound+Imaging+for+Assessment+of+Spinal+Cord+Blood+Flow+in+Experimental+Spinal+Cord+Injury.">Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury.</a> Jove-Journal of Visualized Experiments. (99), e52536 (2015).</li><li>Kuliga, K. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamics+of+Microvascular+Blood+Flow+and+Oxygenation+Measured+Simultaneously+in+Human+Skin.">Dynamics of Microvascular Blood Flow and Oxygenation Measured Simultaneously in Human Skin.</a> Microcirculation. 21 (6), 562-573 (2014).</li><li>Li, Z. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post+pressure+response+of+skin+blood+flowmotions+in+anesthetized+rats+with+spinal+cord+injury.">Post pressure response of skin blood flowmotions in anesthetized rats with spinal cord injury.</a> Microvascular Research. 78 (1), 20-24 (2009).</li><li>Muck-Weymann, M. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory-dependent+laser-Doppler+flux+motion+in+different+skin+areas+and+its+meaning+to+autonomic+nervous+control+of+the+vessels+of+the+skin.">Respiratory-dependent laser-Doppler flux motion in different skin areas and its meaning to autonomic nervous control of the vessels of the skin.</a> Microvasc Res. 52 (1), 69-78 (1996).</li><li>Stefanovska, A., Bracic, M., Kvernmo, H. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wavelet+analysis+of+oscillations+in+the+peripheral+blood+circulation+measured+by+laser+Doppler+technique.">Wavelet analysis of oscillations in the peripheral blood circulation measured by laser Doppler technique.</a> Ieee Transactions on Biomedical Engineering. 46 (10), 1230-1239 (1999).</li></ol>

Alcuni dettagli dovrebbero essere notati durante l'esecuzione di questo protocollo. In primo luogo, il processo di anestesia e chirurgia dovrebbe essere effettuato in modo più rapido ed elegantemente possibile per ridurre al minimo lo stress introdotto all'animale. Per ridurre il disturbo ai risultati, è possibile mantenere l'animale in uno stato relativamente tranquillo e stabile. In secondo luogo, si dovrebbe prestare attenzione più al sanguinamento durante la misurazione utilizzando apparecchiature Doppler laser, p...

Il tessuto del midollo spinale è estremamente sensibile all'ipossia indotta da lesione del midollo spinale (SCI) e altamente vascularized. Nostri studi precedenti hanno mostrato che il flusso di sangue del midollo spinale è stato diminuito significativamente dopo trauma cranico lesioni1,2, che potrebbe essere collegato al deficit della funzione motoria. Recenti studi hanno dimostrato che l'integrità dei vasi sanguigni dopo SCI è ben correlato con il miglioramento della funzione motoria sensoriale3. È stato segnalato che una migliore vascolarizzazione potrebbe salvare la materia bianca, che indirettamente conduce a miglioramento della funzione4. Di conseguenza, il mantenimento della perfusione di alberino-ferita del midollo spinale è sembrato essere di primaria importanza per preservare la vitalità e funzionalità.
Gli effetti dei vari trattamenti su aspersione dopo SCI sono stati esaminati da numerosi ricercatori usando una varietà di tecniche in modelli sperimentali di SCI5,6,7. Laser Doppler, come una tecnica ben consolidata, era senza dubbio un metodo utile per la quantificazione di aspersione in parecchi studi animali ed umani8,9,10,11. La tecnica si basa sulla misurazione l' effetto Doppler12 indotta da globuli rossi in movimento alla luce illuminante. Dalla commercializzazione della tecnica nei primi anni 1980, grandi progressi compiuti nella tecnologia laser, fibre ottiche ed elaborazione dei segnali per aspersione di misurazione laser Doppler strumenti13, che ha reso LDF in una tecnologia affidabile.
Nello studio corrente, entrambi i metodi del laser Doppler misura sono stati applicati per valutare il flusso sanguigno (BF) nei midolli spinali dei ratti concussive. A causa della natura non invadente della tecnologia e sua installazione semplice, il nostro protocollo fornisce un metodo sensibile, rapido e affidabile per misure di BF del midollo spinale. Ancora più importante, questo metodo consente uno studio longitudinale di BF post concussiva SCI senza sacrificio animale in ogni momento.
Grazie alla capacità di valutare il BF del tessuto e veloci cambi di aspersione durante la stimolazione, è possibile applicare questo protocollo per valutare cerebrale BF14,15 , nonché di misurare altri tessuti quali fegato16, 17, pelle18,19e viscere20. In un modello del ratto dell'occlusione transitoria dell'arteria cerebrale centrale, le letture di Doppler del laser sono state utilizzate per assicurare la corretta riduzione del tasso di BF a livelli che sono attesi nella penombra ischemica14. In ratti che sono stati sottoposti a induzione (CLI) l'ischemia critica degli arti, scansione Doppler laser è stata applicata per osservare dell'arto BF prima e dopo la procedura CLI e durante diversi periodi dopo trattamento21. Inoltre, la biodisponibilità e la clearance metabolica di alcuni farmaci dipendeva dal BF epatica, che è stato rilevato da LDF16. Di conseguenza, LDF potrebbe essere ampiamente usato nella valutazione del modello sperimentale, di farmacodinamica e farmacocinetica.

<table>
	<tbody>
		<tr>
			<td>Laser Doppler Line Scanner</td>
			<td>Moor Instruments</td>
			<td>moorLDLS2</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Doppler Monitor</td>
			<td>Moor Instruments</td>
			<td>moorVMS-LDF</td>
			<td></td>
		</tr>
		<tr>
			<td>Probe for Monitor</td>
			<td>Moor Instruments</td>
			<td>VP3</td>
			<td>Blunt needle end delivery probe</td>
		</tr>
		<tr>
			<td>Impactor</td>
			<td>Precision Systems and Instrumentation</td>
			<td>IH-0400</td>
			<td></td>
		</tr>
		<tr>
			<td>Phenobarbital sodium</td>
			<td>Sigma-Aldrich</td>
			<td>P3761</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Sigma-Aldrich</td>
			<td>B-908</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe</td>
			<td>Becton Dickinson Medica (s) Pte.Ltd</td>
			<td>300841</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical suture needles with thread</td>
			<td>Shanghai Pudong Jinhuan Medical Products Co., Ltd</td>
			<td></td>
			<td>18T0329 (batch number) /4-0</td>
		</tr>
		<tr>
			<td>Scalpel</td>
			<td>Operation instrument factory of Shanghai Medical Instrument Co., Ltd.</td>
			<td>J11030 4#</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel blade</td>
			<td>Operation instrument factory of Shanghai Medical Instrument Co., Ltd.</td>
			<td>J12130 20#</td>
			<td></td>
		</tr>
		<tr>
			<td>Ophthalmic forceps</td>
			<td>Operation instrument factory of Shanghai Medical Instrument Co., Ltd.</td>
			<td>JD1040</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemostatic forceps</td>
			<td>Operation instrument factory of Shanghai Medical Instrument Co., Ltd.</td>
			<td>J31050</td>
			<td></td>
		</tr>
		<tr>
			<td>Benzyl penicillin sodium</td>
			<td>North China Pharmaceutical Co., Ltd</td>
			<td></td>
			<td>F6072116 (batch number)</td>
		</tr>
		<tr>
			<td>75% alcohol</td>
			<td>Dezhou Anjie Gaoke disinfection products Co., Ltd</td>
			<td></td>
			<td>150421R (batch number)</td>
		</tr>
		<tr>
			<td>Iodine</td>
			<td>Shandong Lierkang Medical Technology Co., Ltd</td>
			<td></td>
			<td>20170102 (batch number)</td>
		</tr>
		<tr>
			<td>Rat</td>
			<td>Laboratory Animal Center, The Academy of Millitery Medical Sciences</td>
			<td></td>
			<td>Sprague-Dawly (rat strain)</td>
		</tr>
	</tbody>
</table>

using laser doppler imaging and monitoring to analyze spinal cord microcirculation in rat

Flussometria del laser Doppler (LDF) è un metodo non invasivo per la misura di portata (BF) di sangue, che lo rende preferibile per la misurazione microcircolatorie alterazioni del midollo spinale. In questo articolo, il nostro obiettivo era di utilizzare sia Laser Doppler imaging e monitoraggio per analizzare il cambiamento di BF dopo la ferita del midollo spinale. Sia il laser scanner di immagini Doppler e il sonda/monitor sono stati impiegati per ottenere ogni lettura. I dati di LDPI fornito una distribuzione locale di BF, che ha dato una panoramica di aspersione intorno al sito di lesione e reso accessibile per l'analisi comparativa di BF tra posizioni diverse. Intensamente misurando l'area sondaggio per un periodo di tempo, una sonda combinata è stata utilizzata per misurare simultaneamente la saturazione di ossigeno e BF del midollo spinale, mostrando perfusione generale del midollo spinale e rifornimento di ossigeno. LDF stessa ha alcune limitazioni, quali flusso relativo, segnale di sensibilità al movimento e zero biologico. Tuttavia, la tecnologia è stata applicata in studio clinico e sperimentale, grazie alla sua semplice installazione e misura rapida di BF.

LDPI fu utilizzato per misurare il BF nel midollo spinale, che è stato quantificato lungo l'asse rostro-caudale del midollo spinale tramite l'estrazione di profili lineari (Figura 4). Figura 5A e 5B figura rappresentano rispettivamente l'imaging di flusso del midollo spinale del gruppo SCI e del gruppo finto. Figura 5 e Figura 5 rappresenta...

Watch this Scientific Journal Video about Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat at JoVE.com

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Qui presentiamo una combinazione della perfusione Doppler laser imaging (LDPI) e monitoraggio di laser Doppler aspersione (LDPM) per misurare spinale cord flussi sanguigni locali e saturazione dell'ossigeno (SO2), così come una procedura standardizzata per l'introduzione del midollo spinale trauma da ratto.

Utilizzo di Laser Doppler Imaging e monitoraggio per analizzare il microcircolo del midollo spinale nel ratto

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory ...

The overall goal of this procedure is to present a combination of laser doppler imaging and monitoring to measure spinal cord local blood flows and oxygen saturation, as well as a standardized procedure for introducing spinal cord trauma on a rat. This method can help answer key questions in the field of micro circulation such as how blood flow redistributed over time and its impact on the surrounding area after spinal cord injury. The main advantage of this technique is by using both laser doppler imaging and monitoring device we could rapidly derive data of the local distribution of the blood flow as well as the blood flow variation over a period of time, which allows more sophisticated analysis of the blood flow in the problem area.Begin by checking the anesthetized rat for a surgical plane of anesthesia by the absence of a reaction to a toe pinch. Shave the dorsal area of the rat from the lower back to the neck. Then place the rat dorsal side up on a 40-degrees Celsius heating pad to maintain a constant body temperature.Sterilize the shaved area by wiping with cotton balls soaked in iodine followed by 75%alcohol. After making a skin incision over the laminectomy site covering thoracic vertebrae T7 to T11, cut the attached muscles on both sides from T8 to T10 to expose the spinous processes, the laminae, and the facet joints. Use the scalpel to make incisions that disconnect the junction between T10 and T11.Further expose the junction by carefully dissecting the muscle layer away to expose the bone. Use the scissors to further clear muscle away from the lamina and around the pedicle with small snips. This will open a small space between the vertebrae at T10 and T11.Slowly and delicately insert hemostatic forceps into this gap and break the pedicle. Repeat on the other side. Expose the spinal cord and carefully lift and break off the lamina without leaving free or jagged bone fragments behind.Repeat the process to further remove T9 and T8 laminae. An impactor may now be used to induce concussion injury in experimental groups. To scan the exposed spinal cord, place the rat dorsal side up on a black, non-reflective background, then set up the scanning parameters in the software by clicking Measure to enter the measurement graphical user interface and then the Scanner Setup button to open the Scanner Setup interface.To scan small areas such as in this experiment, click Scan Size and Display options and select High Resolution for a fine scanning mode with higher resolution. Click on the Image Scan option to check the scanning parameters. Check the live video image by clicking on the Video and Distance option.Position the scanner 10 to 13 centimeters above the surgical window and move the background with the animal to center the exposed spinal cord on the scanning window. Select the Auto Distance function to fine adjust the scanning height, which should be kept consistent across all measurements in the experiment. Use a non-reflective cover with a window to expose only the surgical area to further minimize background and mark the animal's direction.Click on Repeat Scan to set the number of scans, then click Okay to open the Repeat Scan interface. Click the Start button to start scanning. The whole process will take approximately three to four minutes.A scanner monitor with a VP3 blunt needle and delivery probe is used here to monitor blood flow and oxygen saturation over time. Begin by attaching the probe perpendicular to a stereotaxic instrument to set up the monitoring equipment. Put the rat on the stereotaxic apparatus, dorsal side up, and underlay the animal with a small piece of Styrofoam to level the exposed spinal cord.Examine the incision and remove any excessive liquid or blood using a cotton pad. Use the apparatus's x-and y-axes to locate the probe to two millimeters rostral to the center point of the exposed spinal cord or lesion point and devoid the central vein. Use the z-axis to slowly lower the probe to the level just touching the surface of the spinal cord.The probe should just touch the surface of the spinal cord but not be so loose as to allow bright light to escape from the side of the contact point. Proper positioning is critical for adapting and implementing the monitoring protocol. The probe should be perpendicular to the measured surface and excessive pressure should be avoided.To record data, click on the New Experiment button in the software to open the setup interface. Under the General option, check the system configuration, then click Next. In the Display setup, select the channel for blood flow and oxygen saturation and click Next.Input file information and click Next to enter the data recording interface. Click on the green triangle button to start recording data from the probe. Once the signal is stable, record data for eight consecutive minutes, then lift the probe and remove the animal from the stereotaxic apparatus.After suturing the muscle and the skin, place the rat on its side in a clean cage on a heating pad, avoiding contact between the surgery site and the cage bottom. Monitor the animal until it wakes up from anesthesia to ensure no post-surgery bleeding and that the sutures remain closed. These images represent the flux imaging of the spinal cord of the sham group on the left and the spinal cord injury group on the right.Flux imaging demonstrated that spinal cord injury induced reduction of blood flow and blood flow of the epicenter was lower than rostral cord and caudal cord. These images show the alternating blood flow along the rostral-caudal axis of the spinal cord of the sham group on the left and the SCI group on the right. As shown here, spinal cord blood flow of the spinal cord injury group significantly decreased compared to the sham group.Simultaneously, the oxygen saturation of the spinal cord was remarkably lower after spinal cord concussion, which was consistent with the change in blood flow after injury. After watching this video, you should have a good understanding of how to measure blood flow of the spinal cord using both laser doppler perfusion imaging and the laser doppler perfusion monitoring. Once mastered, the surgery of introducing spinal cord injury can be done in 20 minutes and the measurements, including use both laser doppler instruments, will take roughly 20 minutes if it is performed properly.While attempting this procedure, it's important to remember to use a non-reflective cover with a window to expose only the surgical area to further minimize background and mark the animal's direction. This technique paved the way for researchers in the field of microcirculation to explore the changes of blood flow for its pharamacodynamics and pharmacokinetics evaluation.

Research

JoVE Journal

Behavior

How to Detect Amygdala Activity with Magnetoencephalography using Source Imaging

Come rilevare amigdala attività con Magnetoencefalografia utilizzando Imaging Source

In trace fear conditioning a conditional  stimulus (CS) predicts the occurrence of the unconditional stimulus (UCS),  which is presented after a brief ...

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Comportamento

Measurement of Fronto-limbic Activity Using an Emotional Oddball Task in Children with Familial High Risk for Schizophrenia

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Adolescence is a critical developmental period where the early symptoms of schizophrenia frequently emerge. First-degree relatives of people with ...

Uncovering Beat Deafness: Detecting Rhythm Disorders with Synchronized Finger Tapping and Perceptual Timing Tasks

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A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here ...

Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

Misure comportamentali e locomotore Utilizzo di un campo aperto Sistema di Monitoraggio Attività per le Malattie muscolo scheletrico

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing ...

Using Fiberless, Wearable fNIRS to Monitor Brain Activity in Real-world Cognitive Tasks

Utilizzando Fiberless, fNIRS indossabili per il monitoraggio attività cerebrale in compiti cognitivi del mondo reale

Functional Near Infrared Spectroscopy (fNIRS) is a neuroimaging technique that uses near-infrared light to monitor brain activity. Based on neurovascular ...

The Other End of the Leash: An Experimental Test to Analyze How Owners Interact with Their Pet Dogs

L'altra estremità del guinzaglio: un Test sperimentale per analizzare come i proprietari interagiscono con i loro animali domestici cani

It has been suggested that the way in which owners interact with their dogs can largely vary and influence the dog-owner bond, but very few objective ...

Treating Low Back Pain in Failed Back Surgery Patients with Multicolumn-lead Spinal Cord Stimulation

Il trattamento di dolore lombo-sacrale in pazienti non riuscito ambulatorio posteriore con stimolazione del midollo spinale Multicolumn-piombo

Failed back surgery syndrome (FBSS) refers to persistent, chronic pain following spinal surgery. Spinal cord stimulation with dorsal epidural leads can be ...

Paradigms of Lower Extremity Electrical Stimulation Training After Spinal Cord Injury

Paradigmi della formazione di stimolazione elettrica dell'arto inferiore dopo la ferita del midollo spinale

Skeletal muscle atrophy, increased adiposity and reduced physical activity are key changes observed after spinal cord injury (SCI) and are associated with ...

Simultaneous Recordings of Cortical Local Field Potentials and Electrocorticograms in Response to Nociceptive Laser Stimuli from Freely Moving Rats

Registrazioni simultanee di potenziali di campo locale corticale ed Electrocorticograms in risposta a stimoli nocicettivi Laser da liberi di muoversi

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and ...

Modified Blood Collection from Tail Veins of Non-anesthetized Mice with a Vacuum Blood Collection System and Eyeglass Magnifier

Raccolta del sangue modificate dalle vene di coda dei topi Non anestetizzati con un sistema di raccolta di sangue sotto vuoto e la lente di ingrandimento per occhiali

Blood sample collection is the basis of experimental animal research. It is of importance to obtain adequate blood samples for various research purposes. ...