Can anyone advise on techniques to allow imaging of actin and
microtubules/tubulin in living (macrophages) ? I have a user who wants to look at
bacterial infection of the macrophages, but for various reasons does
not want to adopt the fix, permeabilise and immunostain protocol...
mainly because they would also like to do pH measurments.
[They, but I guess that means we, need to try and follow events over
2-3 days, which include the labelling of the cells, challenge by
bacteria, how/if  they regulate pH  and whether they survive.]
 
Staining for ACTIN:  I have read/heard that one can load cells with
labelled-phallotoxins - by microinjection or scrape loading - and
still retain cell viability. Has anyone on the list done something
similar in sensitive cells like macrophages ?
[We are thinking of trying to load in a solution containing DMSO.. ]
 
Staining for TUBULIN: any ideas here ? Does anyone know whether we
could approach this by loading (again how ?) stained tubulin dimers.
 
I must confess that we haven't tried anything yet, except for the pH
measurements.
 
Comments and/or references for the above would be greatly
appreciated. Thanks
 
Ian.
 
 
 
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Dr Ian Harper
Experimental Biology Programme
Medical Research Council
PO Box 19070              Tel: 027-21-938 0347
Tygerberg 7505            Fax: 027-21-938 0456
South Africa
Internet: [log in to unmask]
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Microscopy Society of Southern Africa
           Web Page at
http://www.uct.ac.za/depts/emu/mssa
 
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