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JoVE Encyclopedia of Experiments
Microbiology
磁铁矿嵌入细菌纳米纤维素球膜的生成
磁铁矿嵌入细菌纳米纤维素球膜的生成
Encyclopedia of Experiments
Microbiology
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Encyclopedia of Experiments Microbiology
Generation of Magnetite-Embedded Bacterial Nanocellulose Pellicles

磁铁矿嵌入细菌纳米纤维素球膜的生成

Protocol
155 Views
05:15 min
November 7, 2025
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Please note that some of the translations on this page are AI generated. Click here for the English version.

Transcript

以细菌纳米纤维素薄膜为例,这是一种胶状纤维素薄膜。

将其放入一个三颈烧瓶中,瓶内有铁盐溶液。

装一个冷凝器,并用水流过,以防止加热时体积流失。然后通过入口用氮气清洗烧瓶,以防止铁氧化。

加热并搅拌烧瓶,在硅油浴中促进铁盐扩散到膜中。

缓慢加入碱基以提高pH值,使铁盐在片膜基质内形成磁铁矿纳米颗粒。

降低温度并继续搅拌,以确保纳米颗粒分布均匀。

冷却混合物后,转移到干净的容器中。

使用磁铁固定胶膜,并倒出未掺入的物质。

用脱氧水反复清洗膜膜,直到pH值变为中性,上清液看起来清澈。

转移胶膜并用紫外线消毒,然后将其浸入无菌脱氧水中进行进一步研究。

用1000毫升高纯度的水和氮气泡泡,去除水中溶解氧并用氮气替代。

使用三颈圆底烧瓶,以2比1摩尔比例的六水化铁(III)和四水化铁(II)四水合物混合,并用脱氧高纯度水稀释。利用容器的两个颈部,通过将氮气供应连接到插入鼻中隔塞并固定在容器颈部的针头上,从而保证氮气的持续进出。将一层BNC膜放入反应物容器中。

确保样品完全浸没在液体中。然后,将所有玻璃接缝注入真空润滑脂。将容器剩余的颈部连接到一个冷凝管,管上装有充满无水硫酸钙的干燥管。

把水从冷凝管里流过。接着,将溶液在硅油浴中加热至80摄氏度,使用搅拌热板保持该温度。用小型磁性搅拌棒以350转/分钟混合反应物5分钟。

确保BNC充分浸渍了铁质溶液,反应物完全溶解。将搅拌速度提高到700转/分钟。在五分钟内,用穿孔塞子的移液针向10毫升亚铁溶液中加入5毫升氢氧化铵。加入氢氧化铵后,溶液颜色从黄橙色变为黑色。

继续以80摄氏度搅拌溶液五分钟。避免高速搅拌以保持样品完整性。用搅拌热板的温控底部将溶液温度降至30摄氏度,继续搅拌五分钟。

然后,关闭电热板。此时,氧化铁纳米颗粒已被纳入BNC网格中。接着,将混合物冷却至室温,转移到容器瓶中,用强永久磁铁分离磁性纳米颗粒(MNPs)和BNC。

为此,将磁铁靠近容器,以固定MNP和BNC,同时倒出上清液。将MNPs和BNC重新浸泡在100毫升水中。轻轻摇晃溶液,去除所有未被BNC强力掺入的MNPs。

然后再次倾倒上清液,使用磁铁固定MNPs和BNC。用水多次清洗MNP和BNC,直到上清液达到中性pH值,使用比色试纸测量。用镊子将磁功能化BNC(MBNC)与MNP分离,并多次用水冲洗MBNC直到水变清。

通过让MBNC在紫外线下过夜消毒。将MBNC储存在20毫升高纯度的脱氧高纯水中,水温在120摄氏度高压灭菌20分钟中。在无菌情况下,将样品浸入1%的聚乙二醇(PEG)中,并在室温下搅拌两小时。PEG涂层提高了沉积在BNC中的氧化铁纳米颗粒的生物相容性和稳定性,因为这些纳米颗粒通过MBNC 3D网络分布。

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