Lumafluor逆向示蹤熒光微球Retrobeads研究?jī)?nèi)側(cè)前額葉mPFC和上丘SC解決方案

前額葉皮層（Prefrontal Cortex, PFC）對(duì)上丘（Superior Colliculus, SC）的調(diào)控如何影響先天逃跑行為的研究，如何使用Lumafluor的fluorescent tracer microspheres逆向示蹤熒光微球標(biāo)記研究，可以參考這篇文獻(xiàn)。

論文信息：

論文題目：Prefrontal control of superior colliculus modulates innate escape behavior following adversity

期刊名稱：Nature Communications

時(shí)間期卷：15, Article number: 2158 (2024)

在線時(shí)間：2024年3月9日

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

產(chǎn)品信息：

貨號(hào)1：R180

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

貨號(hào)2：G180

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

規(guī)格：100ul

品牌：Lumafluor

產(chǎn)地：美國(guó)

名稱：fluorescent tracer microspheres

現(xiàn)貨授權(quán)：Target Technology（靶點(diǎn)科技）

Lumafluor逆向示蹤熒光微球Retrobeads研究?jī)?nèi)側(cè)前額葉mPFC和上丘SC解決方案

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

通過注射酮-甲苯噻嗪溶液 （0.04 ml/20 g，ip） 麻醉所有小鼠，并固定在立體定位框架 （Kopf Instruments） 中。異氟醚 （1%） 由低流量麻醉輸送系統(tǒng) （Kent Scientific） 輸送，以在手術(shù)過程中保持深度麻醉狀態(tài)。通過使用熱探針和自動(dòng)控制器系統(tǒng)控制體溫，將體溫保持在 37 °C。小鼠接受鎮(zhèn)痛劑 Carpofen （0.01 ml/20 g， i.p.） 注射，并在小鼠眼睛上涂抹眼膏 （Duratears），以保護(hù)角膜在手術(shù)過程中不干燥。手術(shù)后，小鼠連續(xù) 4 天 （每 48 小時(shí)注射 1 次） 接受全身性抗生素 （Amoxy LA，0.04 ml/20 g，ip），包括手術(shù)當(dāng)天，并允許小鼠在行為實(shí)驗(yàn)開始前恢復(fù)至少 1 周。根據(jù)具體實(shí)驗(yàn)，注射/植入光電極、光纖、病毒載體或熒光示蹤劑（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.

為了解剖追蹤 SC 傳入神經(jīng)和 DMS 傳入神經(jīng)，將綠色和紅色熒光示蹤微球（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） 稀釋儲(chǔ)備液，每個(gè)進(jìn)樣面積的體積為 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.