How many XFe-96 cartridges ?

In the updated XFe-96 cartridge well plate set-up below, we can compare between the age groups and genders between healthy controls and Parkinson's patients for study 1, as well as running a duplicate on the same plate.

This offers the advantage of both initial experiment and confirmation run being done under the same experimental conditions. 

One of the issues yet to be understood is the expectations for the number of runs required due to the variability or accuracy of the XFe-96 results, ie how many technical references are needed for this type of experiment - is one duplicate sufficient or do we need to run another plate to increase the testing from two cases to four cases ?

The good news is that if we need further plates the costs seem to be affordable if we assume we can buy the extra cartridges from the Binger laboratory rather than a bulk order from Agilent

Study 1 - Bioenergetic changes ...

The first study will look at analysing changes in bioenergetic levels between Parkinson disease participants (PD)and healthy controls (HC) across differing age groups and by gender.

Initially in this study the levels of mitochondrial oxidative respiration will be measure using a Seahorse XFe-96 to simultaneously measure the oxygen consumption rate (OCR) and changes in glycosylation via the Extracellular Acidification Rate (ECAR).

Both oxygen and glucose are necessary in the production of ATP, so by measuring the OCR & ECAR we can estimate the ATP level of production. By measuring OCR & ECAR for healthy individual over the two age groups (40-50 & 60-70 yrs.), we can set up a benchmark to identify how mitochondrial bioenergetic levels change as individuals age.

These levels can be then used as a benchmark for comparison to the same measures in Parkinson disease participants as they age and also between gender.

Research Studies slowly coming together ...

Overall the focus of my PhD research will be on changes in mitochondrial bioenergetics and how the lack of available energy in the form of adenosine triophosphate (ATP) is linked to a wide variety of disease pathologies. 

Specific Studies : 

1. Impact of mitochondrial bioenergetic dysfunction/failure on aged Parkinson’s Disease patients
2. Identify impaired bioenergetic signalling pathways in PD 
3. Proteomic analysis of specific pathway changes in protein abundance
4. Investigate the viability of Leukocytes as a bio-marker of Parkinson’s Disease