Therapeutic Irrigation - Science topic

Hello-

I have been doing BAL fluid collections on mice, with the intention to extract RNA from the cell pellet. I know there are many ways to do this, but the optimal method I landed on is to use 3mls of PBS, one at a time. I've been lavaging/washing each ml 3 times before finally aspirating it and collecting it in a tube. With this method, I was able to get a big enough pellet to get sufficient RNA from a single naive mouse, and have been able to consistently replicate that.

The next step was doing this on mice that were treated with something that causes lung inflammation. I've tried it twice and both time, the RNA yield was significantly lower. I did notice that more of the PBS ended up in the thoracic cavity, so I imagine that's where the cells went and why I was unable to get more RNA.

So, my questions are: is it likely that I didn't do the lavage gently enough, for damaged lungs? Should I limit each ml to one or two washes, or use smaller amounts at a time? Do I need to dispense and aspirate the PBS more slowly? How can I maximize my cell yield in this situation?

Thank you-

Amy

Hello. The question I wanted to pose to you is concerning in vivo assays. One of the possibilities of assays is a bronchoalveolar lavage (BAL). This technique entails que lavage with PBS (for example) of the airways, and collect that fluid, and further analyse it (proteins, enzymes and cells).

In your expertise, can we store the liquid from BAL? Like freezing, for example.

Or do we have to analyse right away?

Do you have any advice on how to tackle this?

Thank you all for your help in advance!

How to analyse the synovial fluid collected from rats. Have collected the synovial fluid lavaged with PBS for 3 times. Now isn`t that concentration of drug in synovial fluid get diluted?If yes, whats the dilution factor to be applied and used for multiply the concentration attained in the synovial fluid post MS analysis.?

Thanks in advance.