I SUPPOSE IF GROWN WITH prdm 16 A BROWN ADIPOCYTE SWITCH CAN BE GENERATED to obtain enough brown adipocytes as the 32c develop from a common precursor myf5 although not white adipocytes which have a digfferent lineage .

If you want to measure the total amount of fat in the muscle, there are standard methods for this. You can extract the fat from the saponification process and suspend the fat by gravimetric method. However, differentiate the fatty acid by ELISA I've never heard.

Dear All,

Thank you for your responses. What standard methods are you referring to? We currently quantify fat and muscle fat radiologically but are interested in extending this understanding. Can you refer me to good protocols for the saponification and gravimetric methods?

Many thanks

Distinguishing between lipid droplets inside muscle syncytia and adipocytes interchelated between muscle fibres is essential for molecular studies of 'intramuscular adipose tissue'. Both may be present in the calf muscle, at least in fairly obese people, along with much aponeurosis and other tendinous material. But standard biopsies are probably too small to rely upon being able to distinguish the two sources.

Dear Alexander,

I agree with Caroline Pond. In fact, standard biopsies are too small to evaluated lipid content by chemical methods.  For the implementation of the technique I mentioned earlier is necessary 1g of tissue. Follows the reference used:

Stansbie D, Brownsey RW, Crettaz M, Denton RM. (1976). Acute effects in vivo of anti-insulin serum on rates of fatty acid synthesis and activities of acetyl-coenzyme A

carboxylase and pyruvate dehydrogenase in liver and epididymal adipose tissue of fed rats. Biochemical Journal 160, 413–416.

Perhaps quantify fat is simpler than qualify. Try non-invasive methods such as magnetic resonance imaging or ultrason.

Sorry for not helping much.

Dear All,

Thank you for this welcome input. In brief, I want to be able to conduct cytokine assays from IMAT. From what I gather, this may not be possible due to the limited amount of tissue that will be harvested from the biopsy?

We already conduct DXA and pQCT to quantify fat non-invasively.