real-time cerebral blood flow monitoring with periorbital doppler probe in rodent middle cerebral artery occlusion stroke models

<meta charset="utf8"></head><body>眶周激光多普勒血流测定法治疗啮齿动物 MCAO 诱导的缺血性卒中

The middle cerebral artery occlusion (MCAO) method has been widely used in rodents since introduced to the scientific community for application to rats in 19861 with the Longa adaption described in 19902, and adaptations for mice soon following3. Although not described in Longa&#39;s publication, the use of laser Doppler flowmetry (LDF) signal to confirm the filament placement was soon described in the literature4. LDF employment during the MCAO procedure is prominently featured in the literature but is designated as an optional step in the current Stroke Preclinical Assessment Networks (SPAN) Standard Operative Procedures (SOPs)5.
The use of LDF confirms correct filament placement during the MCAO procedure and, therefore, contributes to study design rigor and subsequent results, particularly in experiments designed to explore drug efficacy. LDF use reduces surgical mistakes stemming from incorrect filament placement, which results in the dichotomous situation of either no injury when the filament is not placed far enough or animal deaths due to vessel perforation occurring when the filament is inserted too far. On the other hand, the use of LDF is not associated with infarct size variability commonly observed following the MCAO procedure6. The use of LDF in the MCAO procedure may be perceived as difficult and onerous, particularly in rats, because the skull is thicker than in mice and may require skull thinning prior to LDF placement7,8. Also, dorsal probe placement is often described with some protocols requiring specialized equipment or preparation7,8,9. With any of these barriers, implementation of the LDF to confirm filament placement may not take place.
In this protocol, we describe the placement of the laser Doppler probe at the skull and over the distal region of the middle cerebral artery-a periorbital placement-for the assessment of blood flow during the MCAO procedure in both mice and rats. Our rationale was to develop a procedure that has multiple advantages over some methods reported in the literature7,8,9,10 in that it is minimally invasive, fast, and does not require skull thinning or specialized equipment beyond the laser Doppler probe.

<meta charset="utf8"></head><body>作者聚焦：通过啮齿动物模型中区域脑血流的眶周多普勒监测推进中风研究

啮齿动物模型中激光多普勒探头在大脑中动脉闭塞前眶周放置用于脑血流监测

使用眶周多普勒探头实时监测啮齿动物大脑中动脉闭塞卒中模型中的脑血流

real-time-cerebral-blood-flow-monitoring-with-periorbital-doppler

Research

JoVE Journal

Neuroscience

Real-Time Cerebral Blood Flow Monitoring with Periorbital Doppler Probe in Rodent Middle Cerebral Artery Occlusion Stroke Models

73 次观看. 首先，将准备好并麻醉的动物放在手术台上进行手术。使用棉签，从手术区域的中心开始，以螺旋运动将 Betadine 涂抹在准备好的皮肤上。然后用酒精棉片擦拭该区域。使用小剪刀，在眼角和耳褶之间做一个垂直切口，露出浅筋膜和下面的肌肉。然后使用锋利的镊子和弯曲的剪刀，小心地切开肌肉层，直到到达颅骨。使用镊子烧灼工具，在必要?...

视频: 使用眶周多普勒探头实时监测啮齿动物大脑中动脉闭塞卒中模型中的脑血流

Periorbital Placement of a Laser Doppler Probe for Cerebral Blood Flow Monitoring Prior to Middle Cerebral Artery Occlusion in Rodent Models

Middle cerebral artery occlusion (MCAO) is the gold-standard method for preclinical modeling of ischemic stroke in rodents. However, successful occlusion is not guaranteed by even the most skilled surgical hands. Errors primarily occur when the filament is not placed at the correct depth and include instances of either no infarction or vessel perforation, which can cause death. Laser Doppler flowmetry (LDF) is a reliable technique that provides real-time feedback on regional cerebral blood flow (CBF) during the MCAO procedure. Here we demonstrate a rapid technique for periorbital placement of a laser Doppler probe for measurement of CBF in both mice and rats. Our rationale was to simplify LDF implementation, encouraging widespread usage for improved surgical reliability. The technique eliminates the need for skull thinning and specialized equipment, with placement at the periorbital region rather than dorsal placement, promoting efficiency and ease of adoption. The protocol described here encompasses presurgical preparations, periorbital Doppler probe placement, and post-operative care. Representative results include visual depictions of procedural elements along with representative LDF tracings illustrating successful MCAO surgeries, with instances of unsuccessful filament placement leading to complications. The protocol illustrates LDF in confirming proper filament placement and offers a simplified procedure compared to alternative methods.

A minimally invasive surgical procedure is shown here, which involves placing the laser Doppler probe onto the skull over the distal region of the middle cerebral artery (MCA), a periorbital location suitable for rats and mice, to assess blood flow during transient MCA occlusion.

Middle cerebral artery occlusion (MCAO) is the gold-standard method for preclinical modeling of ischemic stroke in rodents. However, successful occlusion ...