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Dear Alex,
For long term imaging having a heated cage incubator is certainly the most stable option, especially if your cells are sensitive to changes in temperature. Hypoxic physiology can obviously be temperature sensitive depending on what you are interested in. You then have a stage-top incubator for localised gaseous and humidity control. Sometimes people even like to have this heated as well for added thermal stability.
If your purchase includes a microscope, then you can opt for one with a hardware autofocus, all 4 main companies have this, and that removes any issue of focal drift attributable to thermal changes. This means that you could then potentially go with just a stage-top incubator, and drop the cage. However, please note. If using immersion objectives in combination with a stage top incubator (no cage), your objective will be at room-ish temperature, and so acts as a heat sink. If you are concerned about the actual temperature your cells are at, then you need to be aware of this. Stage-top incubators can be set to account for the offset, but I think you should still double check the actual temperature at the coverslip. The added available component to compensate for this is an objective heater. Also note that some immersion objectives are temperature corrected when investigating your purchase.
Finally. If running hypoxic experiments, be sure of the oxygen tensions you want to drop down to. I used to work in a lab where they found available solutions a challenge to drop below a certain value. Sorry, cannot remember the finer details at this moment in time. They did find a solution in the end.
Not sure this really answers any questions, but hopefully provides more information with which to make a decision.
I have run stage top incubators with hardware autofocus in overnight experiments and held focus, though was less certain about temperature stability on the cells themselves! Personally, I would go with a cage incubator.
Best
Geraint
Slightly commercial. Have you considered spinning disk confocal technology?
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Alex Paredez
Sent: 24 July 2012 22:35
To: [log in to unmask]
Subject: [CONFOCALMICROSCOPY] Full environmental chamber vs stage top incubator
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Hello,
I am looking into a DeltaVision microscope. The sales rep wants to sell me an environmental chamber which controls temp and CO2. I work on an intestinal parasite (Giardia) that needs a hypoxic environment. The sales rep would like me to put a smaller chamber inside their larger cabinet and control atmospheric conditions with a gas mixer. He argues that by heating the entire stage and all the objectives there will be less thermal drift and better temperature control. I don't know what the gas system is going to cost yet but list on the Environmental chamber is around 11K. I can get a complete stage top system from Pathology Devices for around 5K more than the chamber. I like the idea of not having a giant box in the way when we are imaging fixed samples. Also I am not sure how easy it is going to be to load a smaller chamber with samples when it is inside of a larger cabinet. I could use some advice. I am also not set on the DV or any particular environmental system. That said my department has plenty of confocal microscopes and I think deconvolution is probably the best solution for my small cells.
Thanks,
Alex
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