Preparation of Diplonemid Samples for Microscopy

Area of Science:

• Microscopy and cell biology

Background:

• Diplonemids are marine stramenopiles with complex cellular structures that are challenging to study using traditional microscopy techniques.
• Understanding diplonemid cell biology requires robust and adaptable protocols for high-resolution imaging.

Purpose of the Study:

• To provide a comprehensive set of standardized protocols for preparing diplonemid cells for various microscopy applications.
• To enable researchers to obtain high-quality imaging data for the cellular biology of diplonemids.

Main Methods:

• Detailed protocols for slide preparation, cell harvesting, chemical fixation, and permeabilization.
• Instructions for fluorescent labeling, including immunofluorescence assay (IFA) and fluorescence in situ hybridization (FISH).
• Adaptable methods for live-cell imaging and fixed-cell fluorescence microscopy.

Main Results:

• Established a suite of reliable protocols for diverse microscopy techniques.
• Highlighted critical steps to ensure successful cell preparation and high-quality imaging.
• Demonstrated the adaptability of protocols for different experimental objectives.

Conclusions:

• These protocols facilitate advanced imaging of diplonemid cells, advancing the study of their cellular biology.
• The standardized methods will aid researchers in exploring complex cellular processes in diplonemids.
• High-quality imaging data is achievable through meticulous adherence to these preparation techniques.