Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear Mr. Kozubek,
nice to hear from you again.
I would like to give some answers and explanations in
response to your email.
It is right that the confocal CGS OptoLine is a complete
system which includes the OptiGrid technique developed by
Tony Wilson (University of Oxford).
I would like to extend the list you gave us
System Technique Light_Throughput Light_Source Imaging
--------------------------------------------------------------------------
CARV Nipkow_disk 5% Mercury_lamp Direct
CSU-10 Nipkow_disk+Microlenses up to 40-60% Laser Direct
CSU-10 Nipkow_disk+Microlenses 4% Mercury_lamp Direct
OptoLine Single_frequency_grid up to 65-75% Mercury_lamp Indirect
OptoLine Single_frequency_grid up to 65-75% Halogen_lamp Indirect
--------------------------------------------------------------------------
your Point 6)
The OptoLine aquires a 1.3 MPixel confocal image/sec. It
depends also on the integration time of the camera. If you are using a
camera which is not so light intense it might take longer.
So this means OptoLine is at least as fast as the fastest
full frame CLSM at MPixel resolution.
Your point 7)
The algorithm is not unpublished! The confocal image is formed by
the algorithm which was already published by Tony Wilson in his
original article.
Your point 8)
The resolution of the confocal images captured with the OptoLine
system depends on the resolution of your microscope
objectives. The images are not processed by any additional
steps. The algorithm mentioned in point 7) is simply subtractive.
Your point 9)
During the demonstration we made together I was using an
earlier hardware and software version of the OptoLine
system.
If anybody would like to see the results of CGS OptoLine they can
contact us for a demonstration.
mfg / regards
Anneliese Schmaus
Product Manager
klughammer gmbh
Strassbach 9
85229 Markt Indersdorf
Germany
Tel. +49 08136 6011
Fax +49 08136 7098
www.klughammer.de
MK> Search the CONFOCAL archive at
MK> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
MK> On Tue, 5 Mar 2002, Philip Oshel wrote:
>> First, is the CGS Optoline "confocal" the Optigrid under another
>> name? If I try to link to the Optigrid site, I get Klughammer
>> instead, and a tiny bit of information on this Optoline.
>> Second, does anyone have valid email addresses for Klughammer Gmbh?
>> Their web site barely works, and trying to send them email results in
>> a "server not found" error.
>> Or, does anyone have any information on the CGS Optoline?
MK> Dear list,
MK> CGS Optoline is an alternative to the other cheap confocals such as
MK> CARV (Atto) or CSU-10 (Yokogawa) (distributed mainly by Perkin-Elmer
MK> as Ultraview). All three systems are at approximately the same price
MK> level (about $40000 exclusive microscope, light source and camera).
MK> All three systems use a CCD camera as the detector and are easily
MK> attached to any microscope (in principle). The following table
MK> summarizes the main differences:
MK> System Technique Light_Throughput Light_Source Imaging
MK> --------------------------------------------------------------------------
MK> CARV Nipkow_disk 5% Mercury_lamp Direct
MK> CSU-10 Nipkow_disk+Microlenses up to 40-60% Laser Direct
MK> OptiGrid Single_frequency_grid up to 65-75% Mercury_lamp Indirect
MK> --------------------------------------------------------------------------
MK> Note that the light throughput values are taken from the manufacturers'
MK> specifications, i.e. the real values may be lower (especially in the
MK> case of the very clever "up to" prefixes :-)
MK> Note also that it is possible to use mercury_lamp light source with
MK> CSU-10 but then the light_throughput reduces to about 4%
MK> (see the technical note by Watson-Juskaitis-Wilson in the
MK> latest (February) issue of the Journal of Microscopy).
MK> Concerning the OptiGrid and OptoLine:
MK> -------------------------------------
MK> 1) OptoLine is a complete system based on OptiGrid,
MK> i.e. OptiGrid is a part of OptoLine
MK> (OptoLine = OptiGrid + Driving_Electronics + Software)
MK> 2) OptiGrid (similarly to CARV and CSU-10) can be mounted
MK> in principle to any commercial microscope.
MK> 3) OptiGrid is a trademark of Optem, Inc. and it is sold
MK> by Klughammer (Germany) in Europe and Triptar (US) in America.
MK> 3) The web pages are:
MK> www.klughammer.de (equals to www.optigrid.de)
MK> www.triptar.com
MK> 4) Principles of operation are shortly explained at:
MK> http://www.triptar.com/optigrid.htm (section Principles of Operation)
MK> 5) The idea is based on the principle developed in Tony Wilson's
MK> group in Oxford. The original article is:
MK> Neil MAA, Juskaitis R, Wilson T
MK> "Method of obtaining optical sectioning by using
MK> structured light in a conventional microscope"
MK> OPT LETT 22: (24) 1905-1907 DEC 15 1997
MK> Abstract: We describe a simple method of obtaining optical sectioning
MK> in a conventional wide-field microscope by projecting a
MK> single-spatial-frequency grid pattern onto the object.
MK> Images taken at three spatial positions of the grid are processed
MK> in real time to produce optically sectioned images that are
MK> substantially similar to those obtained with confocal microscopes.
MK> Subsequent modifications:
MK> Neil MAA, Juskaitis R, Wilson T
MK> "Real time 3D fluorescence microscopy by
MK> two beam interference illumination"
MK> OPT COMMUN 153: (1-3) 1-4 JUL 15 1998
MK> (interfering two beams)
MK> Wilson T, Neil MAA, Juskaitis R
MK> "Real-time three-dimensional imaging of macroscopic structures"
MK> J MICROSC-OXFORD 191: 116-118 Part 2 AUG 1998
MK> (macroscopic system)
MK> Neil MAA, Squire A, Juskaitis R, et al.
MK> "Wide-field optically sectioning fluorescence
MK> microscopy with laser illumination"
MK> J MICROSC-OXFORD 197: 1-4 Part 1 JAN 2000
MK> (laser illumination)
MK> 6) All these articles describe systems, in which the image is formed
MK> indirectly in computer memory based on 3 images with a different
MK> grid positioning. This means that for each fluorochrome and
MK> for each z-position you have to acquire 3 images! This implies
MK> that the specimen must be stable and must not move during
MK> these 3 acquisitions! Therefore, this technique is not very
MK> suitable for live cell imaging.
MK> 7) The image is formed using UNPUBLISHED algorithms!!!
MK> You have to believe the software developers that
MK> what they compute is true!
MK> 8) It is very hard to assess the resolution of the system
MK> because you do not know what image processing steps
MK> the software performs.
MK> 9) Mrs. Schmaus from Klughammer was so kind and acquired images
MK> of our FISH-stained microscope slides. Although she was very
MK> kind and willing to repeat the acquisitions, the resulting
MK> images computed by the software looked a bit strange and
MK> artificial. Especially the DAPI-stained cell nuclei had
MK> some strange black holes inside and other parts of the
MK> nucleus were set to maximum intensity level (as if overexposed).
MK> The images looked very similar to what you can find on
MK> the Klughammer web page in the section Fluorescence images
MK> (the blue cell in the third image). I do not think this
MK> is what the software should generate.
MK> 10) Although I have worked in Tony Wilson's group for a year
MK> and highly appreciate their inventions, I was quite
MK> disappointed by the OptiGrid image results. I think
MK> the idea is good but there is some bug in the software
MK> or some other technical problem with this system.
MK> Regards,
MK> Michal
MK> ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
MK> Michal Kozubek, Ph.D.
MK> Laboratory of Optical Microscopy, Faculty of Informatics,
MK> Masaryk University, Botanicka 68a, CZ-60200, Brno, Czech Republic
MK> Tel/Fax/Ans: +420-5-41512467 E-mail: [log in to unmask]
MK> Internet home page: http://www.fi.muni.cz/~kozubek
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