Seahorse XF Flex Analyzer
The Agilent Seahorse XF Flex analyzer measures and reports the oxygen consumption rate (OCR), proton efflux rate (PER) or extracellular acidification rate (ECAR), as well as ATP production rates in a 24-well format. It offers optimized workflows and dedicated consumables to enable real-time metabolic analysis with 3D study models, such as organoids and tissue materials. It also provides wide signal dynamic range through the use of dedicated consumables.
OCR, PER, and ATP production rates are critical attributes of energy metabolism and vital indicators of mitochondrial health, toxicity, glycolysis, and overall cellular (dys) function. The ability to analyze these parameters in more complex three-dimensional structures will reveal deeper insights and enhance the translatability of your research.
The XF Flex analyzer is also equipped with advanced software and advanced data analytics available in the Agilent Seahorse Analytics software. These features greatly simplify the entire XF assay experience, from assay design to data QC and interpretation.
特性
- A flexible 24-well system that allows real-time metabolic analysis with improved sensitivity and consistency, using two-dimensional (2D) and three-dimensional (3D) study models
- Multiple parameters reported from each assay well, including OCR, PER or ECAR, and ATP production rates
- Tailored workflows and dedicated consumables for working with organoids and tissue materials
- Wide detection range and great for high respiration study models
- Four-port injection system with automated mixing feature for assessing immediate cellular, tissue organoid responses to substrates, inhibitors, and other compounds in real time
- Verified instrument performance to ensure intraplate and interplate data consistency
- Broad operation environment temperature of 4 to 30 °C, allowing the analyzer to maintain internal assay temperature of 16 to 42 °C
- Intuitive, user-guided software for easy assay protocol creation
- Automated data quality check in data analysis software to flag abnormal assay behavior
- Data files compatible with advanced data analysis features in the Seahorse Analytics which is a secure web-based data analysis tool
性能指標(biāo)
工作原理
實時監(jiān)測微孔板中的活細胞生物能量代謝
線粒體呼吸和糖酵解這兩個主要的能量產(chǎn)生途徑,分別涉及細胞耗氧量和質(zhì)子釋放率。 Seahorse XF 技術(shù)使用無標(biāo)記傳感器檢測這些分析物中的細胞外變化,以測定細胞呼吸率、糖酵解和 ATP 產(chǎn)生。該技術(shù)可用于研究培養(yǎng)細胞、懸浮細胞以及組織、類器官和/或多細胞簇等三維材料的代謝變化。
形成微室,并計算每分鐘細胞外流量速率
儀器將探針板降低至檢測孔中。傳感器位于孔底上方特定距離處,形成非常小的瞬態(tài)微室,這種設(shè)計對實現(xiàn) Seahorse XF 技術(shù)獨有的高靈敏度檢測至關(guān)重要。 隨著氧氣和 pH 水平的變化,儀器可記錄傳感器中的相應(yīng)變化。測量時間通常為 3 分鐘,并自動執(zhí)行速率計算。每次測量結(jié)束后,升高探針,使細胞外培養(yǎng)基恢復(fù)到基礎(chǔ)條件。
最多注入 4 種化合物,實時測試響應(yīng)或研究生物學(xué)機理
探針板還配置有加藥口(每孔 4 個),可在分析過程中將調(diào)節(jié)因子注入細胞孔中。當(dāng)完成儀器方案配置后,系統(tǒng)會將化合物“A”注入分析孔中,緩慢混合,確保化合物在分析培養(yǎng)基中均勻分布。所有孔以此方式同步處理。系統(tǒng)將自動執(zhí)行后續(xù)測量周期、方案規(guī)定的任何額外加藥及速率計算。
應(yīng)用
探索細胞代謝的強大功能
安捷倫 Seahorse XF 平臺可實時測量活細胞的兩個主要代謝通路(線粒體呼吸和糖酵解),提供細胞生物能量代謝的功能動力學(xué)測量。了解生物能量參數(shù)如何提供有價值的信息,并作為疾病模型、關(guān)鍵細胞過程和療法發(fā)現(xiàn)的指標(biāo)。
免疫代謝
包括激活、增殖和記憶細胞發(fā)育在內(nèi)的免疫細胞過程都是由代謝重編程驅(qū)動的,代謝重編程可以被調(diào)節(jié)以增強性能和控制免疫細胞結(jié)局。通過功能性實時代謝測量,了解激活、增殖和記憶細胞發(fā)育等免疫細胞過程。
癌癥代謝
新陳代謝是癌癥惡性腫瘤細胞生長的關(guān)鍵驅(qū)動因素,為了總體上向糖酵解表型轉(zhuǎn)換,癌細胞增殖通常需要進行上調(diào)或“代謝轉(zhuǎn)換”,從而加快能量需求并生成結(jié)構(gòu)單元,最終促進癌細胞生長。通過對活細胞進行實時功能性生物能量代謝分析,揭示癌癥代謝特性,更深入地了解癌細胞生物學(xué)。
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