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Liu;

It is just optics.

For example for a 1.4 NA objective the Z resolution is 1.7 of the X-Y resolution; therefore, an object will never appear to be as small in Z as it is in X-Y.

This problem is exacerbated if you have spherical aberration caused by refractive index mismatches.

Regards,

Tom

 Tom Donnelly          Applied Precision, LLC
 Biotechnology Group   1040 12th Ave. N.W.
 (425)313-4549         Issaquah, WA 98027-8929
 (425)557-1055 fax     [log in to unmask] http://www.api.com/products/bio/deltavision.html

  

-----Original Message-----
From: Zhiqian Liu [mailto:[log in to unmask]]
Sent: Sunday, May 05, 2002 9:27 PM
To: [log in to unmask]
Subject: Cuticle thickness measurement


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Dear list members,


I am currently measuring leaf cuticle thickness using the vertical section
images acquired from CLSM (either using reflection mode or after staining
with a lipophilic fluorescent dye). The problem I am having is that the
thickness values I get this way is far greater than those obtained with TEM
for the same plant species and growth stage. Does anyone have experience
with similar situations?

I would appreciate your ideas and comments.


Liu
====================
Zhiqian Liu, PhD
Scientist
Forest Research
Private Bag 3020
Rotorua, New Zealand
Tel:  64 (0)7 343 5886
Fax: 64 (0)7 343 5811