What would you advise for in-vivo two photon calcium imaging between Fura-2 and Oregon Green 488-BAPTA? I cannot do ratiometric analysis because the set-up does not allow it. Any suggestions?
Our lab and most labs doing similar work use OGB-1 AM (http://products.invitrogen.com/ivgn/product/O6807) for our in vivo imaging in the cortex for two photon. The signal to noise ratio is good, as is the signal strength for excitatory neurons. Inhibitory neurons are a bit trickier since the calcium buffering in those cells tends to be higher. Astrocytes will also be labelled by OGB-1 AM in vivo, so you may need to combine it with SR101 loading to differentiate between neurons and astrocytes depending on what you are doing.
Eh..the fun thing is that i am doing live imaging on honey bee...i d love to look at the interaction between astrocytes and neurons..but sr101 does not work so well in insects...but i m getting to it somehow..
Ha, that's right, I remember discussing that with you before. But definitely try OGB-1AM first.
Hi,
Fura 2 is not as good in vivo in mammals brain as in vitro. Don't know why... anyway as in any two photon recording the best answer is try it in your own conditions/model and let see !
Dear Marca,
for sure you did it already, but just check the in vivo work by Arthur Konnerths group in Munich. You will find a lot of excellent studies using OGB1 in vivo.
Kind regards
http://www.ifn.me.tum.de/new/
http://www.ncbi.nlm.nih.gov/pubmed?cmd=search&db=pubmed&orig_db=pubmed&term=Konnerth+A.
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