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<DIV><FONT face=Arial size=2>TOMADO DE <A title=ALGAE-L@LISTSERV.HEANET.IE
href="mailto:ALGAE-L@LISTSERV.HEANET.IE">ALGAE-L@LISTSERV.HEANET.IE</A></FONT></DIV>
<DIV><FONT face=Arial
size=2>************************************************************************************</FONT><BR></DIV>Hello
All! <BR><BR>Apparently, the attachment didn't work for most. I am just going to
paste the text into the body of the email. Apologies!<BR><BR>I had a number of
people request that I post responses to my request for information on Hardcover
KLB 2:1-4 and "the perfect microscope and imaging system for diatom microscopy",
so here is my best attempt at condensing the wealth of information I received.
<BR><BR>Thank you to everyone who responded, I greatly appreciate and respect
your opinions. I'm sure this information will help a lot people. Please see
pasted document below for condensed responses concerning KLB, microscopes, and
imaging systems.<BR><BR><U><B>Concerning the hardcover KLB
references:</B></U><BR>A number of people suggested checking <A
href="http://Amazon.com">Amazon.com</A>, Ebay, and Koeltz. Perhaps if
I<BR>continue to check back I may happen across a set being sold used, but
currently, to my<BR>knowledge and those that responded, there is no supplier
that has a stock of hardcover<BR>copies.<BR>A number of people also made
suggestions, such as having the paperback books spiral<BR>bound or bound in
hardcover. This is a great idea, but the paperback copy that I saw<BR>also
seemed to have a lower quality, non-glossy paper, reducing the quality of
the<BR>photomicrographs in the plates. Hopefully this is something we can
resolve in the future.<BR><BR><U><B>Concerning Microscopes:</B></U><BR>In
general, a number of things were recommended by many:<BR><B>1.</B> Differential
Interphase Contrast or DIC (and/or phase contrast as a
cheaper<BR>alternative)<U> to increase contrast</U><BR><B>2. About
Objectives:</B><BR><B>a. </B>The best available 100x oil immersion objective,
generally 'Apochromatic'<BR>(has Numerical Aperature or NA >1.4), <U>to
increase resolution</U>. In order to use<BR>DIC with this objective you will
also need to purchase both an interference<BR>contrast objective prism and a
condenser prism.<BR><BR>As a note: this one objective may potentially be the
single most expensive<BR>piece on the microscope ($5000.00 US or more), but is
also one of the most<BR>important pieces of the microscope. It was suggested by
many, that every<BR>effort should be made to get a 100x objective with an NA of
1.4 or greater<BR>(best if matched to the NA of the condenser) even if it meant
it was the only<BR>objective you could afford to purchase for the
scope.<BR><BR><B>b.</B> Although apochromatic is best, because the 100x
objective is used most<BR>frequently for identifying diatoms, in order to save
money, a 'Semi-<BR>Apochromatic' 40x or 60x objective was suggested (this should
cost<BR>significantly less, but is still somewhat expensive). Also, a number of
people<BR>recommended having the 60x objective be oil immersion as well
(<U>this<BR>increases resolution)</U>. Again, in order to use DIC with this
objective, you will<BR>also need to purchase both an interference contrast
objective prism and a<BR>condenser prism.<BR><BR><B>c. </B>5x, 10x, and 20x
objectives may be useful, but are not generally necessary to<BR>identify diatoms
to the species level. If these objectives are desired a simple<BR>"Achromatic"
objective can be purchased. For many companies, you could<BR>get all three (5x,
10, and 20x) for under $1000 US. Generally, for diatoms, it<BR>is not necessary
to have the DIC slider or prisms for any of these objectives.<BR><BR><B>d.</B>
The final suggestion for objectives is that a diamond scribe (aka
diamond<BR>stylus) is purchase for the nosepiece position. This will allow you
to circle<BR>specimens (so you can find them later) as well as draw lines across
the<BR>coverslip (to aid in counting). This is helpful for a number of reasons:
1. You<BR>can create a personal reference collection by circling species, then
be able to<BR>easily locate them in the future. 2. Circling specimens also
allows for easier<BR>collaboration. You can circle a specimen, then send it to a
colleague for<BR>verification, or help with identification. 3. It is also
helpful for QA/QC (Quality<BR>Assurance). For example, if you have a line drawn
across the coverslip, you<BR>can easily re-count a slide along the same path
used before.<BR><BR><B>Conclusion: </B>Start with the best 100x objective you
can afford, fill in the rest<BR>when you can. Remember that you can almost
always, unless they are<BR>discontinued, choose to add on objectives and prisms
at a later date. As you<BR>move from lower quality to higher quality objectives,
you will have fewer<BR>aberrations, essentially increasing resolution to the
eye.<BR><BR><B>3. About the Condenser:</B><BR><B>a.</B> The NA (1.4) of the
substage condenser should match the NA of the<BR>objective, in order to get the
best <U>resolution</U>.<BR><B><BR>b.</B> Many suggested that the substage
condenser should be oil immersion (just<BR>as the objective is). This will
increase resolution. Essentially, an oiled<BR>condenser allows an oil immersion
interface between the slide and the<BR>condenser main element.<BR><B>4. About
the stage:</B><BR><B>a.</B> Some complained of faulty specimen holders, so be
sure that the specimen<BR>holder is of a good quality and very
sturdy.<BR><B><BR>5. Illumination:</B><BR><B>a.</B> DIC creates a "light starved
environment", so be sure to have, at the least,<BR>high wattage (100w) halogen
illumination. To upgrade to a tin halide<BR>illumination, it is VERY expensive
($5000.00 US or more), so it is mostly<BR>unnecessary. But be sure to replace
bulbs often to get the best images.<BR><BR><B>6. About specific
manufacturers:</B><BR><B>a. Leica </B>I had the most recommendations from those
who owned Leica<BR>microscopes. Many have the Leica DMLB, which is now
discontinued, but the<BR>Leica DM 2500 appears to be comparable. All of those
who wrote to me<BR>about Leica were very happy with the quality of the scope.
One person<BR>warned not to get a scope with preset illumination or programming
because<BR>you have better contrast and options when setting this manually. I
recently<BR>demoed a Leica scope and really appreciated the ergonomic features
they<BR>have added. As diatomists, we often spend hours in front of the scope
and<BR>these features made it much more comfortable. For example, the
eyepieces<BR>could be shifted up and down, allowing you to look straight-forward
rather<BR>than down into the scope. The focus knobs and stage movement knobs
are<BR>very low on the scope, allowing you to comfortably rest your hand on
the<BR>table and reducing wrist/arm fatigue. Also, you can switch the
stage<BR>movement from left to right in seconds by using a magnet. As a note:
other<BR>companies are advertising some of these ergo benefits as well, I just
haven't<BR>had the chance to try them yet. <B>Bottom Line: Leica offers high
quality<BR>scopes and optics at a good price.</B><BR><B><BR>b. Zeiss</B> Nearly
as many responded about Zeiss scopes as did about Leica, and<BR>they were all
very positive. More than one person said that the Zeiss optics<BR>cannot be
beat, but they are often a bit more expensive. The only complaint<BR>about Zeiss
was that they only had a one-year warranty (compared to as<BR>much as 5 years
elsewhere). If you aren't investing in automated stages and<BR>other
pre-programmed equipment, this probably isn't a problem, though.<BR><B>Botttom
Line: Zeiss offers what some say are the highest quality optics<BR>and great
scopes, but it may cost you a bit more.</B><BR><B><BR>c. Nikon</B> A few people
wrote about Nikon scopes, all were happy with the<BR>quality of the optics. The
current recommended scope is the 80i (manual) or<BR>the 90i (automated).
Apparently there were a number of glitches with the<BR>automated stage during
the first month, but all problems were solved with a<BR>bit of back in forth.
Now the scope comes highly recommended. Nikon was<BR>said to have a 5 year
warranty, which could prove to be helpful, especially if<BR>you choose to go
with automated parts. <B>Bottom Line: Nikon also offers<BR>high quality scopes
and optics, but fewer researchers seam to end up<BR>with these scopes.<BR><BR>d.
Olympus </B>A couple of people also wrote about Olympus scopes. Both had
the<BR>BX51 with DIC, one also had phase contrast. Both were extremely happy
with<BR>their scopes. According to others that wrote, Olympus used to be
regarded as<BR>not quite as good as other companies, but they are apparently
stepping up to<BR>the plate in recent years. Bottom Line: Olympus may not have
always had<BR>the highest quality optics for DIC, but more recently are more
than<BR>capable competing with other companies, providing both high
quality<BR>optics and scopes.<BR><BR><U><B>Concerning Cameras and Imaging
software</B></U><BR><B>1. </B>There is a bit of debate on whether or not a
cooled camera system is necessary or<BR>not. Many have cooled cameras and many
don't. I haven't decided which is best<BR>for our purpose, yet. I do, however,
know that cooled cameras cost significantly<BR>more than uncooled.<BR><B>2.
</B>Although this is coming from knowledge garnered from different dealers and I
don't<BR>completely understand it myself, I thought I would pass along this
information, you<BR>may take it for what it is worth: when taking pictures at
100x, you will not get better<BR>images with a higher megapixel (MP) camera. The
optimal would be a 2MP<BR>camera. It has something to do with the number of
times that the area is "sampled"<BR>and the amount of "noise" you will get in
images if a 8MP camera "over-samples" a<BR>highly magnified image. Like I said,
I'm no expert on this, but the bottom line is that<BR>a 2MP camera is what you
want. If you have information otherwise, please share it.<BR><B>3.</B> I got
very mixed reviews on imaging systems from Zeiss, Leica, Nikon, and<BR>Olympus.
One company, Diagnostic Instruments with SPOT imaging systems<BR>came out on top
in the end.<BR><B>a. Zeiss</B> One user felt quality was okay, while another
said the software was<BR>horrible, difficult to understand and essentially a
waste of time.<BR><B><BR>b. Leica </B>One user said there was certainly a
learning curve with software, so<BR>they export to photoshop to manipulate
photos, but it does the job. Another<BR>user said the software was difficult to
use with erroneous functions that don't<BR>allow you to take the pictures you
want and also that the camera breaks<BR>easily and can be a pain. When I had a
demo here, the system crashed and<BR>we were unable to use any of the functions
of the $5000 US software<BR>package he was trying to sell during the demo. I
took it as a sign to run, if the<BR>Leica dealer couldn't get it to work, I
don't know what I would do.<BR><B><BR>c. Nikon</B> One user has the Nikon DS-U2
5mp with Nikon NIS Elements D<BR>documentation and measuring software, they
consider it to be user friendly<BR>and of good quality. Another user is not
satisfied with images captured with a<BR>DS 5M camera from Nikon.<BR><B><BR>d.
Olympus </B>I only heard from one user that had an Olympus camera, he
was<BR>very satisfied with the camera and the microsuite software.<BR><B><BR>e.
Diagnostic Instruments SPOT </B>I heard from numerous users about how<BR>great
SPOT cameras are. One great thing is that you do not have to<BR>purchase
software separately; it comes free with their cameras. Some have<BR>the much
cheaper, but uncooled Insight 1800 series of cameras. You can<BR>choose to have
these in color or monochromatic. Still others have the much<BR>more expensive,
but higher quality and cooled RT2500 series. Again, you can<BR>choose to have
monochromatic or one with a slider allowing high quality color<BR>images. One
user said that they found the Insight to be more user friendly<BR>and preferred
it to the "better" RT series. Overall SPOT cameras and<BR>software were said to
offer:<BR>i. Great measuring and scale-bar technology<BR>ii. Fast
focusing<BR>iii. Simple brightness/contrast manipulation is simple<BR>iv. Easy
use, with hardly any learning curve<BR>v. Great photos all the time even with
limited computer<BR>knowledge<BR>vi. Photoshop brand software is integrated into
their software<BR>package. Many people are already familiar with its
capabilities.<BR><B><BR>Good Luck with finding the scope of your dreams. I hope
that at least some of this can<BR>help you. It certainly helped
me.<BR></B><BR><BR clear=all> <BR>-- <BR>Emily K. Hollingsworth<BR>Benthic
Phycologist<BR>Aquatic Biology Section<BR>Dept of Health & Environmental
Control<BR>2600 Bull Street<BR>Columbia, SC 29210<BR><BR>Work:
803.898.4331<BR>Cell: 616.516.9901<BR><A
href="mailto:HollinEK@dhec.sc.gov">HollinEK@dhec.sc.gov</A><BR><A
href="mailto:EKBiofilm@gmail.com">EKBiofilm@gmail.com</A> </BODY></HTML>