The imidazole buffer allows the retention of the most difficult intracellular lipids to preserve.
The imidazole buffer allows lipid retention. associated with osmium during fixation as a buffer at 0.1M
here is a standard protocol
- Glutaraldehyde ‘2 to 2.5% in 0.1M imidazole buffer pH7.5 – RT
- Wash in 0.2M imidazole buffer pH7.5 (2 baths 3 mins) in RT
- Osmium 1% to 2% in imidazole buffer 0.1 pH 7.5, 30 min – RT
- Last wash same as point 2- (Wash in 0.2M imidazole buffer pH7.5 (2 baths not more, 3 mins) in RT
Replace your usual Sorensen PBS or cacodylate buffers with imidazole , when your study gives importance to lipid inclusions.
important to note: if you want to preserve the contents of the fixed cells, avoid washing too much, 1 wash is enough, if simple lipids are the most difficult to preserve, washing should even be avoided.