Help with what's what on my new microscope

Here you can discuss different microscopic techniques and illumination methods, such as Brightfield, Darkfield, Phase Contrast, DIC, Oblique illumination, etc.
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NJH
Posts: 13
Joined: Tue Jul 17, 2018 12:50 pm

Help with what's what on my new microscope

#1 Post by NJH » Wed Aug 15, 2018 6:26 pm

Hi All,
I need some help identifying the parts on my new microscope. I got it from a surplus auction; it's a Labophot 2 POL - so....the condenser is definitely marked as Phase - 2; the lamphouse is for a 100w Mercury bulb (although I will need to get a power source). But...what I'm not sure of is 1) do I need to replace the Phase condenser with a regular abbe condenser for regular brightfield viewing? 2) what are the ND2/ND4 filters for (Phase/Fluoro/Darkfield?) 3) what is the shutter slider for - again, only "open" when using fluor/polar/phase???

I have the manual(s) for the Labophot, but this particular scope has "more" than just the Labophot that is pictured in all the manuals/brochures; I need to determine what all I have and what else I need to find!!

Can anyone help with the various "bells & whistles" on this Labophot???


I'll attach a few pics here too...if I can figure out how to do it properly!!!
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Hobbyst46
Posts: 4277
Joined: Mon Aug 21, 2017 9:02 pm

Re: Help with what's what on my new microscope

#2 Post by Hobbyst46 » Wed Aug 15, 2018 7:35 pm

From the photos, I think that your 100W mercury lamphouse is for epi illumination - from the top, down through the objective onto the specimen, then (reflected/scattered/fluorescence) back again up through the objective towards the eyepieces and/or camera. This illuminator is useful for fluorescence and you might use it also for epi-darkfield, under precautions.

The ND2, ND4 filters are two grades of neutral density filters - useful, since the 100W mercury emits very intense light - sometimes too intense even for fluorescence.

The shutter is an ON/OFF manual shutter, to open or block the light beam, respectively. It is useful and important because the life-time of mercury bulbs is relatively short, and is further shortened by frequent turning it on and off. Hence, you want to turn on the lamp and leave it ON for the day (or for the session). During the observations, if you need to block the light beam, push/pull the shutter instead of turning off the lamp.

In fact, the mercury lamp is essential for fluorescence. It can be used for epi-darkfield, but is not optimal - you must use appropriate filters to protect your eyes, that is, to avoid reflected/scatterd UV light back into your eyes. Which filters and dichroic mirrors are installed in your scope? Although, if this scope has been used for fluorescence before, I am sure it has the appropriate filters.

The only POL I know is trans illumination, then again it would not be based on the mercury lamp.

So, if I am not completely wrong, this mercury illumination is not used for brightfield, neither for phase contrast or POL. For those, you need the lower lamphouse, for trans illumination, and it should be halogen or LED or something else - not mercury.

It would be nice to see a photo of the whole setup - microscope. Thanks in advance.

NJH
Posts: 13
Joined: Tue Jul 17, 2018 12:50 pm

Re: Help with what's what on my new microscope

#3 Post by NJH » Wed Aug 15, 2018 10:55 pm

Hobbyist46:

Thanks for the detailed information!! This scope was listed as being used in a Serology Lab and I got it via a surplus auction. The ND filters are fairly easily deciphered (via Google Search) and I figured the shutter was to "close" off the light when needed...glad to know I wasn't too far off on those!!! There are two filter cubes in the scope, but I don't know what excite/emit wavelengths they are...haven't looked far enough (yet) to see if they are marked. Could you tell me about the "Bertrand lens"?? Also, can the phase-2 condenser be used for brightfield...or is this scope not designed for brightfield at all??? (OH NOOOOO...that will break my heart!!) I have a polar objective (I assume, with a "target" on it), two Phase 40X, 1 E 100X oil,
another 4X, 10X, 40X and 100X objectives from my Alphaphot YS scope, and I have an abbe condenser from that scope as well. If you are familiar with this scope...the stage is x/y, the condenser lowering knob doesn't move the condenser down. (there's something that'll need to be fixed too!!) I paid $400 for the scope - do you think it was an okay deal? It looks like I might be able to make that back by piecing it out (if it won't fit my needs) - the objectives, the base, the bi-head, the illumination tube, the lamphouse, filters, cubes...but I'd really like to get it working so that I can enjoy it myself - and take it to my workplace (research lab) and use it for fluorescent stains. I'll attach a picture of the entire unit at the bottom of this post. Any additional speculation that you could add would be much appreciated...I know you can only do so much by photos, but you clearly are very well-versed in microscopes!!
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billbillt
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Re: Help with what's what on my new microscope

#4 Post by billbillt » Thu Aug 16, 2018 12:13 am

LED Light Sources: A Major Advance in Fluorescence Microscopy..

https://microscopy-analysis.com/editori ... microscopy

PeteM
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Re: Help with what's what on my new microscope

#5 Post by PeteM » Thu Aug 16, 2018 1:35 am

Most phase condensers, almost surely including yours, have an open portion (served by an iris) that is designed for brightfield. You should be ready to go on that front.

It isn't entirely clear from your picture - but most Labophots had a 20 watt halogen bulb at the rear of the base for brightfield, darkfield, and phase contrast work. Looks to me like yours does as well; but some of these microscopes were epi only. You could always swap a base from another Labophot or cobble a new illumination system together if, by remote chance, yours doesn't have substage illumination.

Hobbyst46
Posts: 4277
Joined: Mon Aug 21, 2017 9:02 pm

Re: Help with what's what on my new microscope

#6 Post by Hobbyst46 » Thu Aug 16, 2018 7:19 am

You are most welcome, I restrict my answer to what I know from other microscopes, I have used an Optiphot, not a Labophot, but some principles are general.
NJH wrote:...There are two filter cubes in the scope, but I don't know what excite/emit wavelengths they are...haven't looked far enough (yet) to see if they are marked....
These filters and dichroic mirrors are surface coated, and the coating is often very sensitive, I would not touch them with bare fingers, but wear rubber gloves and hold them by the rim if possible.
Could you tell me about the "Bertrand lens"??
AFAIK a Bertrand lens is installed between the nosepiece and the head, and serves as a visual aid (instead of a phase telescope) to align the phase rings of the condenser with those of the phase objectives. The Bertrand lens comes within an Optovar or similar attachment.
...or is this scope not designed for brightfield at all???
It is POSSIBLE to use a fluorescence microscope without brightfield (or any other non-fluorescence illumination) but, for me at least, would be highly inconvenient. So, I speculate that your scope IS designed for BF and has an illumination port in the low rear end, for a second illuminator which would be, I believe, halogen. A photo view of the rear side will help. This second illuminator can be halogen or, alternatively, a LED replacement. The light intensity of a LED illuminator for BF and phase contrast can be much lower than the light intensity for fluorescence. Even more importantly, the former is usually white or warm white light, whereas the color of the latter depends on the application. For example if you wanted to detect a fluorescein stain, you need to excite it at 490-495nm. That means an intense 495nm beam. But, a white or warm white light emits a wide spectrum, so the intensity at a specific wavelength is just a fraction of the nominal intensity of the illuminator. (see below more about LEDs).

BTW, did the phase condenser come with the Alphaphot or was it included with the Labophot?
I paid $400 for the scope - do you think it was an okay deal?
- I do not know Labophot prices, but I think that the answer to your question depends on which features of the microscope scope did you seek in the first place. If your wish was fluorescence, and if the illuminator is working (if you still need a power supply, note that this must be a dedicated PS for that high pressure mercury lamp!), and if the cubes and their components are usable, then the price was reasonable, since such accessories, would by themselves cost much much more than $400. My opinion.
but I'd really like to get it working so that I can enjoy it myself - and take it to my workplace (research lab) and use it for fluorescent stains.
This is a well focused and important purpose! Given that you are familiar with fluorescence, there are some requirements to be met:
1. Dedicated power supply for the 100W Hg illuminator.
2. A working high pressure 100W Hg bulb.
3. Initial alignment of the lamp in the lamphouse.
4. Adjustment and optimization of the illumination on the scope.
5. Appropriate cubes for the specific use in the research lab.

All those subjects are probably covered in the instruction manual of the microscope.

Incidentally, there are LED illuminators for fluorescence microscopes, for example the PE-100 and PE-300 from coolLED (a US firm), that are meant to replace mercury and xenon illuminators. They are sufficiently bright, IMO. If the wavelengths you need in the lab are already known (I believe so), you may want to look for LED illuminators. Then you will not need the mercury illuminator and some of the cube components. Please keep in mind that excitation illumination should be much more intense than BF or phase or illumination (I have not compared excitation intensity with DF intensity though).
Also, commercial LED illuminators for fluorescence microscopy are expensive (very expensive from a hobby aspect, but that is subjective...). Trouble is, that missing parts for the Hg lamp are also expensive... sorry, fluorescence is costly!

I note that the scope has no camera port, you may want to upgrade this as well. Also, if you decide to use it for fluorescence without BF illumination, you will not use the condenser at all, so no need to fix the rack...

P.S. Note, that if the excitation light that you need in the lab is UV (e.g. 365nm), better results are obtained by using UV-specific objectives, that transmit these wavelengths. But this would probably be of lower priority on your list, if at all.

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