Posted by
Stanislav Vitha on
URL: http://confocal-microscopy-list.275.s1.nabble.com/DIC-or-Phase-contrast-for-image-morphometry-mesasurements-tp7068286p7071285.html
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Use blue or green filter for imaging to minimize chromatic aberration.
Brightfield with properly set up Köhler illumination (open the condenser
diaphragm as much as you can while still getting acceptable contrast) should
provide for best resolution without the edge artifacts of DIC or phase
contrast.
With a 100x/1.3 or 1.4 oil immersion objective and a typical interline
transfer CCD camera (6.45 um pixel size), you should be OK as far as Nyquist
sampling.
I find DIC quite tricky for measurements of cell outlines, given the
shadowing effects.
If you are OK with somewhat lower resolution of the phase contrast
technique, you can minimize the phase halo around the cells by increasing
the refractive index of the medium - you could add some BSA (if I remember
correctly, something like 10 % is used by some).
Stan Vitha
On Tue, 6 Dec 2011 14:23:26 -0600, Vasseur Monique
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>Dear All,
>
>I wonder what is better for yeast morphometry shape and size image analysis:
>should we better use phase contrast images or DIC images? Is one more
>accurate for measures? )Which threshold will be more precise: the one of DIC
>shear or the one of phase contrast halo?) Do some of you have experience on
>this question? Any input is welcome Thanks a lot!
>Monique