Hi y'all, it's been a rough year and I've been unsuccessful in most experiments so here I am, seeking help.

I've been trying new treatments to impair my tissue. There's no protocol reference for ex vivo work, so I've been trying to optimise things based on in vitro references. It worked the first 5 times and now there is little to no impairment seen. Even my other treatment, that has been optimised and protocols established in the lab, stopped working and is similar to controls now.

My PIs are getting impatient and honestly so am I. I don't know what went wrong or what's wrong, there are so many additional variables due to ex vivo work. Both my advisors came to look at my results today and my main advisor told me to look into alternative treatments and suggested that I get my C57 mice from another source. My other advisor disagrees and thinks different batches of mice aren't affecting results.

My question is, has anyone had issues with their mice, like physiologically? Our department doesn't breed mice, we get them from our university's animal facility in another building.

I had great results earlier on, when I was just practicing. Now that I'm more experienced, it seems that I had gotten worse at basically everything. It's demoralising. Any help or advice is appreciated, thank you.