Dear Goran, My name is Katarzyna and for over two years now I have been
working in a confocal microscopy lab. The method you describe as vertical
stacking the depth of field is, I assume, confocal microscopy. Thanks to a
system of pinholes out-of-focus light is eliminated and only focal plane is
captured. This allows to capture several levels/stacks (you actually can define
how many stacks you need – the more the resolution is better; or which should
be the step size of each stack ie. 0.1μm)
in the z dimension and later
combine them to obtain 3D image. In our lab we work with Leica Systems (SP5 and
SL), unfortunately I don´t know anybody who would use the 3D images obtained
this way for geometric-morphometrics, but I am pretty sure that it is possible.
Leica SP5 has a z range of 1.5mm, so if your specimens are bigger than that it
can be a problem, also xy dimensions are limited due to the objective
magnification;
however I´m not familiar with Leica DVM system you mentioned, so this can be
different there. I do not know it this is of any help, but please feel free
to contact me if you have any questions about confocal microscopy. Best regards, Katarzyna Górka
----- Forwarded message from Kasia
Górka
Date: Wed, 10 Oct 2012
09:07:02 -0400
From: Kasia Górka
Reply-To: Kasia Górka
Subject: Re: 3D
digital microscopy
To: morphmet@morphometrics.org
Department of Anthropology
Faculty of
Biology
University of Barcelona
----- Forwarded message from Göran Arnqvist -----
Date: Tue, 9 Oct 2012 17:45:52 -0400
From: Göran Arnqvist
Reply-To: Göran Arnqvist
Subject: 3D digital microscopy
To: morphmet@morphometrics.org
Dear all,
I wonder if anyone has experience of 3D digital microscopy for
gathering good quality 3D data for geometric morphometrics? It seems
like 3D systems, like the Leica DVM’s or the Hirox KH’s, should be
able to generate reasonably good 3D surface or landmark data? These
systems are based on the principle of focus variation - the limited
depth of field of the optics is utilized to determine depth
information for the specimen. Vertical movement of the specimen
relative to the objective determines the focus information along with
the distance to the optics. They then create 3D images by vertically
stacking the depth of field, focus point to focus point in the depth
gradient created by a motorized stage, followed by 3D tiling. Hirox
claims a Z “resolution” of 0.05 um and a “repeatability” of 0.5 um.
Leica claims “resolution” 0.5 um. However, these numbers refer to the
resolution of the motorized focusing drive and may or may not reflect
the actual resolution/repeatabbility along the Z-dimension in
reconstructed 3D images. I work primarily with insects (or parts
thereof) in the size range 0.1 – 3 mm. Anyone out there who could
share insights and/or experience of these sorts of 3D microscopy
systems?
Cheers,
/Göran Arnqvist
Email: goran.arnqv...@ebc.uu.se
********************************************************************
Prof. Göran Arnqvist
Animal Ecology
Department of Ecology and Evolution
Evolutionary Biology Centre
University of Uppsala
Norbyvägen 18d
SE - 752 36 Uppsala
Sweden
Email: goran.arnqv...@ebc.uu.se
Phone: +46-(0)18-471 2645
Cell phone: +46-(0)70-2935032
Fax: +46-(0)18-471 6484
NEW homepage with PDF downloads and more at:
http://www.anst.uu.se/goarn789/index.html
********************************************************************
----- End forwarded message -----
----- End forwarded message
-----
2012/10/10 <morphmet_modera...@morphometrics.org>
- 3D digital microscopy morphmet_moderator
- Re: 3D digital microscopy morphmet_moderator
