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 fi...

Full description

Saved in:
Bibliographic Details
Published in:Protist 2022-10, Vol.173 (5), p.125905-125905, Article 125905
Main Authors: Seah, Brandon Kwee Boon, Emmerich, Christiane, Singh, Aditi, Swart, Estienne Carl
Format: Article
Language:English
Subjects:
antibody
Ciliophora
culture
cytoskeleton
Ethanol
Fixatives
fluorescence microscopy
Formaldehyde
Karyorelictea
Methanol
Microscopy, Fluorescence
Nitrates
Soil
Zinc Sulfate
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1434-4610
1618-0941
DOI:10.1016/j.protis.2022.125905