CryoAPEX Technique for Sub Cellular Preservation

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Abstract

Researchers at Purdue University have developed a method of preparing live cells for analysis that achieves reduced morphological damage to cellular membranes and membrane bound organelles. A limitation of current cellular preservation techniques is that in order to make a sample compatible with staining, the membrane of the cell becomes damaged and the cellular architecture is lost. Researchers at Purdue University have created a method (known as cryoAPEX) of preserving cells in a way that combines chemical fixation processes with high pressure freezing of cells with peroxidase tagging for localization of membrane proteins. Unlike previously available techniques, this method can be used on cells grown in tissue cultures. This method also requires fewer steps than currently available methods of cellular preservation and is compatible with electron tomography. This technology has applications in biological sample analysis and the preservation of biological samples. Advantages: – Fewer steps than previously available cellular preservation methods – Prevents damage to cell membranes -Compatible with Electron Tomography – Compatible with cells grown in tissue cultures Applications: – Biological sample preservation – Analysis of membrane proteins Technology Validation: The cryoAPEX technologywas validated to obtain a high-resolution three dimensional contextual map of the human FIC (filamentation induced by cAMP) protein, HYPE (also known as FICD. CryoAPEX analysis shows that, under normal and/or resting conditions, HYPE localizes robustly within the subdomains of the ER and is not detected in the secretory pathway or other organelles Publication: CryoAPEX – An electron tomography tool for subcellular localization of membrane proteins Journal of Cell Science (2019) 132, jcs222315. doi:10.1242/jcs.222315

Advantages

Fewer steps than previously available cellular preservation methods

Prevents damage to cell membranes

Compatible with Electron Tomography

Compatible with cells grown in tissue cultures

Potential Applications

Biological sample preservation

Analysis of membrane proteins

Contact Information

Name: Ayantika Ghosh-Bhattacharjee

Email: aghoshbh@purdue.edu

Phone: 765-588-3825