|
Dear list,
I would like to take a 3D view of the branchial arches of 9.5 and 10.5 dpc mouse embryos. I would need to look approximately 400um into the tissue with three Alexa-conjugated fluorophores + DAPI.
My immunofluorescence protocol works with cryosections and I can cut at pretty much any thickness through the tissue. The confocal microscope I have access to is the Leica TCS SP2.
So, my questions are:
1) What is the maximum depth of a z-stack that can be imaged on this type of system?
2) If I take thick cryosections (say around 100um), and image them individually would I be able to assemble a "stack of stacks" from the resulting data and generate a single, 3D view?
3) More generally does anyone have any tips on how to approach such an imaging task?
Many thanks,
Joe
|
Locked
