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

<meta charset="utf8"></head><body>げっ歯類のMCAO誘発性虚血性脳卒中のための眼窩周囲レーザードップラー流量計

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>著者スポットライト:げっ歯類モデルにおける局所脳血流の眼窩周囲ドップラーモニタリングによる脳卒中研究の進歩

げっ歯類モデルにおける中大脳動脈閉塞前の脳血流モニタリングのためのレーザードップラープローブの眼窩周囲配置

げっ歯類中大脳動脈閉塞脳卒中モデルにおける眼窩周囲ドップラープローブによるリアルタイム脳血流モニタリング

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 ビュー. まず、準備して麻酔をかけた動物を手術台の上に置きます。綿棒を使用して、手術領域の中心かららせん状に準備した皮膚にベタジンを塗布します。次に、アルコールパッドでその部分を拭きます。小さなハサミを使用して、目の角と耳のひだの間に垂直に切り込みを入れて、表在性筋膜とその下の筋肉を露出させます。次に、鋭い鉗子と...

ビデオ: げっ歯類中大脳動脈閉塞脳卒中モデルにおける眼窩周囲ドップラープローブによるリアルタイム脳血流モニタリング

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 ...