Great Tool To Assess Metabolic Activity In Cells

June 29, 2018

Johns Hopkins School of Medicine
Oncology
Graduate Student

Overall

Performance

Ease-of-Optimization

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Company:

Agilent Technologies

Product Name:

Seahorse XFe96 Analyzer

Catalog Number:

Seahorse XFe96 Analyzer

Our lab studies the role of mTOR signaling in T cell differentiation and function. One way of controlling this process is through the metabolic reprogramming of T cells to either a glycolytic or oxidative phosphorylation program. To monitor these states, the seahorse analyzer allows us to determine glycoltyic state through acidification rate (ECAR) and oxygen consumption (OCR) with small amount of cells using the 96 well format. This machine allows rapid and easy convenience to monitor the metabolic state of cells.

Experimental Design and Results Summary

Application

Metabolic State Monitoring

Protocol Overview

Using the 96 well format, we seed the appropriate number of T cells in the plate. We use a DMEM unbuffered media along with glucose, glutamine, and pyruvate. The cells are seeded onto the plates using this media for analysis. Depending on the assay, the media can be adjusted for the assay. We use metabolic inhibitors to block certain parts of the respiration to induce maximal activity of ECAR and OCR for maximal reserve capacity.

Tips

Titrate cell numbers and also drug concentrations depending on assay and type of cells.

Features Summary

Small scale 96 well format allows minimal cell number if rare population of interest.

DOI or PMID #

25893604

Additional Notes

N/A

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Summary

The Good

Really allows rapid assessment of metabolic state of cells.

The Bad

Machine is quite expensive and the cost per experiment can be pricey since the cartridges are expensive.

The Bottom Line

Recommend to labs studying metabolism since it provides a basic assessment of metabolic state. Once you get accustomed to the assay, it is quite straightforward and reliable.

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