Lumafluor逆向示踪荧光微球Retrobeads研究内侧前额叶mPFC和上丘SC解决方案

前额叶皮层（Prefrontal Cortex, PFC）对上丘（Superior Colliculus, SC）的调控如何影响先天逃跑行为的研究，如何使用Lumafluor的fluorescent tracer microspheres逆向示踪荧光微球标记研究，可以参考这篇文献。

论文信息：

论文题目：Prefrontal control of superior colliculus modulates innate escape behavior following adversity

期刊名称：Nature Communications

时间期卷：15, Article number: 2158 (2024)

在线时间：2024年3月9日

DOI：doi.org/10.1038/s41467-024-46460-z

产品信息：

货号1：R180

品名1：Red fluorescent tracer microspheres：Red Retrobeads™ IX

货号2：G180

品名1：Green fluorescent tracer microspheres：Green Retrobeads™ IX

规格：100ul

品牌：Lumafluor

产地：美国

名称：fluorescent tracer microspheres

现货授权：Target Technology（靶点科技）

Lumafluor逆向示踪荧光微球Retrobeads研究内侧前额叶mPFC和上丘SC解决方案

To confirm the presence of DMS-projecting mPFC neurons that also branch to the SC, two different approaches were used. First, 3 animals were injected with green fluorescent microbeads (Retrobeads, Lumafluor) in the DMS and red retrobeads in the SC ( Fig. a, b); the other way around, 2 more animals were injected ( Fig. c, d). This allowed visual evidence of an abundant number of neurons in the mPFC, labeled with both red and green, suggesting that they were bifurcating mPFC neurons. However, because the microbeads aggregated within the cells, no reliable quantification of the number of those cells could be made using this method.

Stereotaxic surgery

通过注射酮-甲苯噻嗪溶液 （0.04 ml/20 g，ip） 麻醉所有小鼠，并固定在立体定位框架 （Kopf Instruments） 中。异氟醚 （1%） 由低流量麻醉输送系统 （Kent Scientific） 输送，以在手术过程中保持深度麻醉状态。通过使用热探针和自动控制器系统控制体温，将体温保持在 37 °C。小鼠接受镇痛剂 Carpofen （0.01 ml/20 g， i.p.） 注射，并在小鼠眼睛上涂抹眼膏 （Duratears），以保护角膜在手术过程中不干燥。手术后，小鼠连续 4 天 （每 48 小时注射 1 次） 接受全身性抗生素 （Amoxy LA，0.04 ml/20 g，ip），包括手术当天，并允许小鼠在行为实验开始前恢复至少 1 周。根据具体实验，注射/植入光电极、光纤、病毒载体或荧光示踪剂（Retrobeads、Lumafluor）。All mice were anesthetized by injection of a Ketamine-Xylazine Solution (0.04 ml/20 g, i.p.) and secured in a stereotaxic frame (Kopf Instruments). Isoflurane (1%) was delivered by a low-flow anesthesia delivery system (Kent Scientific) to maintain a deep anesthetized state over the course of the surgery. Body temperature was maintained at 37 °C by controlling their temperature with a thermal probe and an automatic controller system. Mice received an injection of analgesic agent Carpofen (0.01 ml/20 g, i.p.) and ophthalmic ointment (Duratears) was applied on the mice eyes to protect the cornea from drying during the surgery. Following surgery, mice received systemic antibiotics (Amoxy LA, 0.04 ml/20 g, i.p.) for 4 consequent days (1 injection every 48 h) including the day of the surgery and were allowed to recover for at least 1 week before initiation of behavioral experiments. Depending on the specific experiment, optrodes, optic fibers, viral vectors or fluorescent tracers (Retrobeads, Lumafluor) were injected/implanted.

为了解剖追踪 SC 传入神经和 DMS 传入神经，将绿色和红色荧光示踪微球（Retrobeads、Lumafluor）注射到 SC（AP：-3.80 mm，ML：± 0.8 mm，DV：-1.5 mm）和 DMS（AP：0.5 mm，ML：± 1.25 mm，DV-3 mm）中。用 pbs （1/4） 稀释储备液，每个进样面积的体积为 600 μL。

For anatomical tracing of SC afferents and DMS afferents, Green and Red fluorescent tracer microspheres (Retrobeads, Lumafluor) were injected into the SC (AP:−3.80 mm, ML: ± 0.8 mm, DV:−1.5 mm) and into the DMS (AP: + 0.5 mm, ML: ± 1.25 mm, DV-3mm). The stock solution was diluted in pbs (1/4) and the volume per injection area was 600 μL.