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Dear Mr. Moreno,

The OptiGrid converts the illumination system of a conventional 
widefield microscope
to a structured light illumination system. Control of Grid movement and 
image
processing of the raw structured illumination images by a computer 
system allow for
the generation of images with only in focus detail (Neil et al 1997). 
This technique
effectively confers confocal capabilities to conventional wide field 
microscopes.
from: Thales-Optem brochure "Grid intensity study"

OptiGrid is a different approach to gaining confocal images than 
deconvolution or any other software solution.
It is a hardware solution in combination with software.

When planning to use OptiGrid it is a very important point whether you 
want to capture fluorescence
images at different wavelengths or at a single wavelength.

The reason is the chromatic aberration:
A lens will not focus different  colors in exactly the same place 
because the focal length depends on
refraction and the index of refraction for blue light (short 
wavelengths) is larger than that of red light
(long wavelengths). The amount of chromatic aberration depends on the 
dispersion of the glass.
**
That means if you want to capture a confocal image with OptiGrid with a 
DAPI, a GFP and a
Texas Red filter you will capture images from different planes in the 
object. When you will do an
overlay of all three images (DAPI, GFP, Texas Red)  you will get a nice 
image but of three different
z-levels of the object. Depending on field size, problems may increase

This is also relevant in confocal microscopy where pinhole misalignments 
typically manifest themselves
between different bands with very bad effects.

The solution for this problem is the OptoLine ViCo confocal system which 
our company is selling and which
was developed and is still improved by Mr. Pier Alberto Benedetti.

OptoLine ViCo is based upon structured light illumination. ViCo OptoLine 
allows capturing many different
fluorescence ranges from one image.  The motorised spatial modulator 
corrects the chromatic aberration.
For OptoLine Vico the misalignment is not a problem since emission 
pinholes are materially absent and the
computer based detection approach used is intrinsecally self-adaptive 
about this aspect. However, as in
conventional fluorescence microscopes, chromatic pixel-shift can 
represent a limitation, especially in specific
cases such as in colocalization studies. In OptoLine ViCo the 
longitudinal or axial chromatic aberration can
be properly corrected focusing the spatial modulator in the excitation 
path (as well as, eventually, the objective
in the emission one).

Finally it can be said that OptiGrid is an easy-to-use confocal system 
for capturing images with a single wavelength.
Images from visible UV to near infrared can be captured. The sales price 
for the system is reasonable. There will be
hardly any future maintenance costs.

I hope this information gave you some additional information.

Please contact me if you have further questions.

Anneliese Schmaus
Product Manager

klughammer bio gmbh
Strassbach 9
85229 Markt Indersdorf
Germany
Tel. +49 (0)8136 6011
Fax  +49 (0)8136 7098
[log in to unmask] <mailto:[log in to unmask]>
www.klughammer.de <http://www.klughammer.de>




 


moreno wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear all,
>
> we are looking for a live imaging system for an inverted microscope.
>
> I used to work with metamorh but after the change in policy after the 
> acquisition from molecular devices, I would radder prefer Image Pro 
> Plus 5. I just receive a letter saying that they (Molecular Devices) 
> will not support version bellow 6.3....the last one. They are 
> definitively looking for money and not user happiness!!!
>
> Together with this software the portuguese rep is trying to sell 
> optigrid as well (price for reference center). I do know that it will 
> take 3 pictures (=>bleaching) for making the "confocality" but in some 
> cases that is not the problem and it can fill a gap in our facility 
> (for long time experiments on embryos it gets much cheaper and easy to 
> operate than a laser scanning system). However, I a bit "arriere 
> pensee" on system performance and overall functionality.
>
> Does any one already experiment this device?
>
> For the moment the desired setup is:
> ImageProPlus 5
> Opigrid
> Hamamatsu Orca AG (similar to ER)
>
> All the best,
> Nuno Moreno
>