I am interested to check ROS as well as energy level of PBMC of same patient at different time point after Liver transplant ,is it okay i make freez down of PBMC and study ROS as well other energy parameter at the end point of my study?
does freezing change the energy parameter of the cells?
how to proceed my experiment ?
This is possible if you will take small samples of the transplanted liver, by freeze clamp method and determine metabolites such as ATP, AD, AMP, adenosine and Pi using HPLC. Freeze clamp is very important. The size of the sample you determine experimentally on rats (not mice). There exist also a method for thick needle sampling. Finally, you can select a biochemical blood test for your purpose to evaluate the liver's function. Glucose and albumin reflect synthetic capacities.
Dear Alexander,
I am interested to study the energy level during liver regeneration after liver transplantation,And my main focus is PBMC at different time point after liver transplant,
So taking tissue sample at singlepoint does not help me.
Thanking you your kind response.
I don't feel its a good choice to freeze down any samples for ROS estimations..as there maybe significant changes in ROS levels during thawing and processing..also to check energy levels..please try checking ATP, NADH, NADPH and ion levels like Ca2+ etc..i suggest doing them immediately or not delaying it beyond 10-12 h after sample collection when snap frozen cryogenically..
- Badges
- Science topic
- Similar topics
- Medicine
- Disease
- Infectious Diseases