荧光显微镜下纤毛虫批量培养和固定的改进方法

IF 1.9 3区生物学 Q4 MICROBIOLOGY

Protist Pub Date : 2022-10-01 DOI:10.1016/j.protis.2022.125905

Brandon Kwee Boon Seah, Christiane Emmerich, Aditi Singh, Estienne Carl Swart

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引用次数: 0

摘要

Loxodes是最具生态学特征的纤毛虫属之一，具有许多有趣的生理能力，包括重力感应细胞器和硝酸盐呼吸。然而，这些细胞被认为难以批量培养，并且用荧光显微镜使用的常规固定液保存得很差。在这里，我们描述了用液体土壤提取物培养基大量生长和收获Loxodes细胞的方法，以及一种名为ZFAE(硫酸锌、甲醛、乙酸、乙醇)的新型固定剂，它比缓冲的甲醛或甲醇更有效地固定Loxodes细胞。我们发现ZFAE与免疫荧光和核染色DAPI兼容。因此，Loxodes现在可以在实验室条件下对单克隆菌株进行长期维护，大规模生长和现代细胞生物学研究。

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Improved Methods for Bulk Cultivation and Fixation of Loxodes Ciliates for Fluorescence Microscopy

Loxodes is one of the best ecologically characterized ciliate genera with numerous intriguing physiological abilities, including gravity-sensing organelles and nitrate respiration. However, these cells have been considered challenging to cultivate in bulk, and are poorly preserved by conventional fixatives used for fluorescence microscopy. Here we describe methods to grow and harvest Loxodes cells in bulk with liquid soil extract medium, as well as a new fixative called ZFAE (zinc sulfate, formaldehyde, acetic acid, ethanol) that can fix Loxodes cells more effectively than buffered formaldehyde or methanol. We show that ZFAE is compatible with immunofluorescence and the nuclear stain DAPI. Loxodes is thus now amenable to long-term maintenance, large-scale growth, and modern cell biology investigations of monoclonal strains in laboratory conditions.

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来源期刊

Protist 生物-微生物学

CiteScore

3.60

自引率

4.00%

发文量

审稿时长

18.7 weeks

期刊介绍： Protist is the international forum for reporting substantial and novel findings in any area of research on protists. The criteria for acceptance of manuscripts are scientific excellence, significance, and interest for a broad readership. Suitable subject areas include: molecular, cell and developmental biology, biochemistry, systematics and phylogeny, and ecology of protists. Both autotrophic and heterotrophic protists as well as parasites are covered. The journal publishes original papers, short historical perspectives and includes a news and views section.