Mitochondrial and Cell Metabolism: Sea Horse XF assays

The Technology

The Sea Horse XF96 extracellular flow analyzer uses solid state fluorescence probes to measure the rate of oxygen consumption (OCR) and extracellular acidification (ECAR) by cells or isolated mitochondria in 96-well plates. These measures are used to delineate the contribution of oxidative mitochondrial and non-mitochondrial energy production to cellular bioenergetics in real time. The automated delivery of pathway-specific substrates and chemical inhibitors/activators of mitochondrial respiratory complexes subunits during the assay gives further insight into the inner workings of the cells. These metabolic strategies are the key to essentially every single cellular process, be it a physiological adaptation to high performance exercise or the highly pathological brain toxicity seen in neurological diseases or the very detrimental cell proliferation seen in cancer.

Inquiries

Melissa Kazantzis: (561) 228-2898

Location

 B148 (Scripps Core Labs Area -Building B ground floor)

Getting It Done

Scientists can obtain a free training to become independent certified users of the Instrument. Contact Melissa for the training. Certified Scripps FL users must pre book the instrument on our Outlook calendar system and operate the instrument according to its posted guidelines.

Rates

  • Scripps Research Community: $125 per assay
  • External Academic: $125 per assay plus indirect costs (plus labor, if applicable)
  • External for profit: $125 per assay plus indirect costs and labor

For Cloud-Based Data Analysis, Click Here.

To Download Desktop SeaHorse Data Analysis Software, Wave by Agilent, Click Here.

Data Analysis: Calculating ATP Production Rate from OCR and ECAR

The Brand group at the Buck Institute for Research on Aging in Novato, California has been a pioneer and key contributor to the development of Sea Horse Bioenergetic assays and data analysis. The group has published extensively on the fundamental biochemistry principles connecting ATP production with OCR and ECAR which form are the basis of its recommendations on Sea Horse data analysis. Along these lines, for meaningful comparisons of metabolic profiles to be attained, OCR and ECAR ought to be both converted to ATP. We adopt such recommendations in our analysis and can assist our users with these calculations. The link below will take you to one of many references from the Brand group on the subject.

Reference: https://www.jbc.org/content/292/17/7189.full.pdf