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  IONOVATION SCOUT 人工脂質膜重組電生理系統

IONOVATION SCOUT 人工脂質膜重組電生理系統

產品概述:


德國Ionovation公司生產的Ionovation scout人工脂質膜重組電生理分析系統,是該領域最新的產品之一,該產品克服了傳統膜片鉗的一系列缺點,無論從產品的技術含量還是從產品的應用領域上來看,在電生理分析技術中始終處于領先的地位,代表著電生理分析技術發展的方向,是國內外細胞電生理分析實驗室首選實驗儀器。這種高度靈活的桌面檢測系統可適應多種實驗條件,目前已經被用于對細胞膜離子通道、多種動物和植物轉運蛋白和它們的細胞器進行研究。

Ionovation scout 技術特點:


√不需要建立千兆歐姆的封阻

√是唯一一種采用雙層封阻的測量方式

√可用于檢測各種細胞膜上的離子通道、囊泡、配體

√容易實現對單個離子通道進行分析檢測

√可以在膜的兩側改變條件,形成雙信道進行分析    

         

技術原理:


Ionovation Scout采用了人工生物脂質膜來構建離子通道的測量方法。該系統使用兩個獨立的電生理測量室進行測量。該雙層室由一個25μm厚、直徑120微米的聚四氟乙烯隔膜隔開(a)。兩個聚碳酸酯小室的體積分別為1.2 ml。聚碳酸酯小室頂部有孔,方便接入的Ag / AgCl電極和注入緩沖液(a)。當兩個腔室中填充有合適的緩沖液時,溶解在正癸烷的脂質被分散到直徑120微米的特氟隆隔膜的微孔( b)中。緩沖液的液面經過連續地降低和升高過程,直到特氟隆隔膜上的過量脂質被除去,特氟隆隔膜上形成脂雙層自組織形成的人工生物脂質膜(C)。 

然后根據具體的目的蛋白或者細胞器等進行操作。比如類似于α-溶血素的毒素,可以直接加入聚碳酸酯小室中進行測試。而需要重組的離子通道則需要將其與脂肪酸結合為重組脂蛋白,然后將這些脂蛋白或者從生物膜上“剪下”囊泡與人工生物脂質膜通過離子梯度或融合肽方法進行融合。

準備好樣品后,開始進行電生理學測試。通過施加不同的電壓,來監測細胞單離子通道的電流,其中在5-10 kHz帶寬的信號會存儲在系統的硬盤上,測試通常采用的模式是電壓鉗模式(voltage clamp mode),這種模式下應用不同的電壓來得到目的信號。

應用領域:


1.脂質觸發器

2.細胞膜轉運處理

3.膜活性劑比如突觸核蛋白

4.藥物安全分析

5.信號轉導模式分析

應用案例


Ionovation Scout測試系統可用于多種離子通道的分析:

1. 測量對象:可實現對上萬種離子通道、轉運體和孔隙活動的精確測量;

2. 軟件:電生理數據采集分析軟件Patchmaster,采用用戶界面友好的軟件實現測量全部控制;具備預定義協議式的用戶定義實驗工作流程;

3. 自動操作:全自動設備操作(雙層生產和驗證、雙層完整性的電容控制、膜雙側灌注系統);雙層分子可視化操作;

4. 電流測量:穩定低噪聲Ag/AgCl電極,采用鹽橋記錄電流范圍從pA到幾個nA

5. 耗材:一次性雙分子層室可簡單、快速置換;

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