I have attempted several times to stain whole leaves of Arabidopsis for
detection of necrosis / micronecrosis. I have used a Trypan Blue method
(in lactophenol), whith which I have vacuum infiltrated, boiled, or just
let the stain absorb into the leaves. With this method I try destaining
with Ethanol (later I'll try chloral hydrate). The leaves stain nicely,
but do not destain (even after more than 4 days), and there is no
detection of necrotic tissue, even with positive control.
The second method I have used consists of staining with Evan's Blue,
rinsing free dye, grinding tissues, and taking the A600 of the
homogenate. Neither of these methods are giving me good, or even
satisfactory, results. If anyone has some experience or suggestions on
these or other methods for staining micronecrosis, please let me know.
Any comments will be very appreciated.
Thank you all in advance.
jaydub at mindless.com