A methodological basis for SEM autoradiography: biosynthesis and radioligand binding.

Area of Science:

• Cell Biology
• Microscopy
• Radiochemistry

Background:

• Scanning electron microscopy (SEM) is crucial for visualizing cell surface topography.
• Autoradiography allows for the localization of biomolecules but can compromise high-resolution imaging.
• Existing methods often struggle with preserving fine cellular details and silver grain integrity.

Purpose of the Study:

• To develop an improved SEM autoradiography technique.
• To enable high-resolution imaging of cell surface-associated molecules.
• To facilitate the study of cell surface receptors and antigens.

Main Methods:

• A novel method for SEM autoradiography was developed.
• The technique involves degelatinating the emulsion while preserving silver grains.
• It allows for the use of secondary, backscattered electron, and X-ray imaging.

Main Results:

• High-resolution cell surface details are successfully retained.
• Emulsion degelatination is achieved without significant loss or redistribution of silver grains.
• The method provides a reliable medium-sized marker for SEM topographic studies.

Conclusions:

• This SEM autoradiography method offers a significant advancement for visualizing cell surface structures.
• It is suitable for studying biosynthesized molecules and cell surface receptors/antigens.
• The technique facilitates detailed topographic analysis using radio-labelled ligands.