We have cleaned the system thoroughly, prepared new buffers and used the control rats. However, the problem remains the same.
Not enough preload. May need to adjust flow or pressure to maintain adequate preload to support the afterload.
Make sure you don't have leaks through the aorta.
You could try a 10-12ml/min flow for rats
Check the temperature of the water bath. Do you work with constant pressure? If so, you should check the perfusion pressure. It would also be useful to check the pH and oxygen levels of the Krebs solution.
Also check the quality of salt you use in your buffers. Did you change providers?
We had this situation for months without being able to know what was wrong.
So... good luck !
Keep us informed If you can successfully find the problem.
We have had similar problem for several months. The main problem was probably in pH of Krebs-Henseleit solutions. Another big issue is temperature, which affects not only the heart itself (contractility), but also pH and amount of oxygen in solution.
You need good nerves to go through this. I wish you good luck!
I don't know what your working model is ? Could you precise and may be i'll be able to answer.
As as I got no answer, If your post charge is high (too high) your organ will be rapidly in jeopardy. Don't forget that besides all factors (cleaning, salts and so on) the oxygen pressure is very high in your bath, but the oxygen content is very very low, so it is not difficult to observe the preparation "stalling" after a few minutes. Remember that any organ perfused with a Krebs solution is by definition ischemic.
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