> Dear confocalists
>
> Like many other confocal users I want to study processes in
> living cells. To do this we use a Zeiss 410 confocal microscope setup,
> however there are no facilities to keep our cells at 37 degrees
> celsius. We can buy from Zeiss a heating plate, but this is a very
> expensive solution. I wonder what methods are used by others to keep
> cells alive. One of the problems we expect by using a heating
> plate is the heat drain from the lens just underneath our
> preparation. How do you measure the temperature ?. We are using cell
> preparations cultured on glass.
>
> All reactions are welcomed.
 
 
We have successfully used a system based on lab-tek chambered
coverglass dishes together with a heating system built by Brook
Industries in Illinois. There is a linearly controlled heated plate
designed for the specific stage on the microscope (they can make
them for any stage. The 8 chambered
dishes also have a metal casing which surrounds the dish while
maintaining contact with the heated base. A small probe can be used
to monitor sample temperature. We have a heater unit from Bioptechs
for the objective, but I find this unit unsatisfactory and so we are
trying to design one that better suits our needs. We currently have
this system operating on a Bio-Rad MRC 1000 UV/vis with a Nikon
300 inverted scope. It is very stable, does not "bounce" around as
the controller maintains a steady voltage to the plate. We do a lot
of calcium, frap, oxidative measurements etc on live cells on a
daily basis using the time course software on the Bio-Rad instrument.
Our assays can be fast (3 mins total for calcium) or quite long (up
to 3 hours for some of the oxidative studies).
 
Our heater unit is from:
Brook Industries is at 25570 Lehmann Blvd, Lake Villa, Il 60046, ph
708 356 1045.
 
 
Paul
 
J.Paul Robinson, Purdue University Cytometry Labs
[log in to unmask] PH:317-494 6449 FAX:317-494 0517
web http://www.cyto.purdue.edu