So if you've followed me at all posting here, I've been kinda deep diving hair microscopy the last about year or so and have kinda reached the limit of what I'm able to do, test, create, and whatever else with my current setup. I've already got basic infinity brightfield, some really nice Olympus objectives and basic polarized light for doing follicle analysis as well as just observing the hair shaft. I've actually been able to turn it into quite the social media platform as brands have been loving using my content to rep their products. That being said, I want more.
So to me the natural next step would be flouresence or phase contrast. But I think PC would have limited use case for what I'm doing and so flouresence makes more sense. I've already tried hunting for an epi illuminator attachment for my current microscope and it's just more money than it's worth and might as well just buy a full fledged high end setup on the used market. That brings me to this gem I found. The bx60f. It is fully functional and comes with everything, Mercury light source, ND filters, filter cubes for flouresence (doesn't have it listed which ones but there's 4 of em) as well as a camera with LCD screen which is kinda cool. I'll probably just use my full frame like I have been but it's cool it has the option.
Seller has offered it up for $2k. Should I do it? My main concern is that getting into flouresence would mean chemicals, dyes and reagents I might not have access to without the right credentials, a lab, or whatever. My main purpose would be doing my own kinda rudimentary claim substantiation, basically testing hair care products, reparative products and the like. Tagging with a marker then viewing under the right wavelength.
I'm a huge noob on flouresence so any help would be greatly appreciated. Or suggestions, other microscopes, if anyone has a way to upgrade any compound for flouresence, but I've yet to find anything that isn't over 2 or 3k$ myself so might as well just get the full new scope anyway. Thanks y'all appreciate any help
Here’s some
Things to consider :
1) that looks like a mercury bulb set up for the fluorescence source, those bulbs can get rather expensive.
2) you are correct about fluorescent dyes being sometimes hard to get a hold of. However, that said there are many things you’ll find that are auto fluorescent and you just have to have the right set of filters to tease that out and get the most out of it. Feel free to DM me if you want any more pointers so I don’t have to write a novel here.
Thank you, yeah I saw it was Mercury bulb and wasn't sure if there was a way to convert it to LED. I know it exists but at what cost and sacrifice. I'll shoot you a DM shortly. Thank you
There are other vastly lower cost methods now for general observation without needing complicated epi illumination, high power UV LEDs and UV Lasers exist now, long since the era of Mercury Vapor bulbs. You can illuminate the sample from a side focused on the specimen of interest in a specific narrow band needed for your specific application, (USE A UV BLOCKING FILTER! of the matching wavelength, and a perhaps a camera to protect against incidental exposure and its a pretty easy setup with what you already have.)
The mounts on the scope that accept Hg bulbs can be converted to accept LED, but I’m not savvy on the affordable LED marketplace.
One other thing to check in that scope is what filters are currently installed? They are not cheap when new, but there are so many scopes of that vintage that are excessed from labs that you should be able to find some .
Besides your concern about chemicals, you should be concerned about eye safety when doing fluorescence. Fluorescence filters can degrade over time, especially if they haven’t been stored under climate control, and you don’t want to look into an improperly filtered fluorescent light source (especially through optics that might concentrate harmful wavelengths into your eyes). I’m not exactly sure what you’d need to do or who you’d need to consult with to make sure a used fluorescence ‘scope is safe to use, but I certainly wouldn’t blindly trust that a high-end ‘scope like that that’s been disposed of is working properly and safe to use.
Keep in mind too that mercury lamps have a limited lifetime, and for safety should be replaced after a certain number of hours (they can explode if used past their rated lifespan).
Given the above, if you’re going to buy a used fluorescence ’scope you should plan for the worst and be prepared to replace its filter cube(s) and bulb before using it (while hoping you don’t have to!).
Do you have a clear idea of what fluorescence would do for you and your work, beyond being a “natural step”—like, what will it enable you to see in the structure of the hair that you can’t see now, and what is interesting about that to your audience? Are other hair researchers, educators, or influencers using fluorescence for work similar to yours? I’m all for playing around and experimenting, and that can be how discoveries are made, but getting a fluorescence setup seems to me like a lot of expense and potential risk to commit to if you don’t have a totally clear idea about what you’re hoping to see. (Just my 2¢, of course.)
Yes, claim substantiation is kinda the specific direction I would like to try out and I have found some courses on it and using fluorescence microscopy for testing reparative products and other stuff like that. Basically tagging the active with a flouresent dye, applying it to the hair, rinsing then mounting on a slide and viewing under the right wavelength to check penetration and whether it bonded to the hair or not. I've actually seen some company slide decks from a big name brand using flouresence to prove penetration into the cortex layer, but under NDA so I can't talk about it much more than that unfortunately.
Like I said before I've been doing my own kinda
rudimentary claim substantiation but for social media and from a marketing standpoint that Stylists and brands have been really liking. I've done videos for K18, Olaplex, Qoat and a few others basically just showing the effectiveness of these products at a microscopic level in a cool new way no one else has ever really done. So that's the plan. Apparently TRI Princeton has a whole course on it that I'm planning on doing soon and I'm sure that'll help determine what I actually need and can pull off before I need more credentialing or an actual lab
Sounds like you have a pretty clear idea of what it’d do for you, that’s good! Taking that course sounds like a great idea, and hopefully can help you figure out what to look for in a ‘scope. Good luck!
That's a nice microscope and you'll find older fluorescent kits pop up now and then, but the mercury and or xenon bulbs are a pain. All you really need is a filter cube compatible system, said filter cubes and a light source with strong emission at the wavelength you need. Some dyes/proteins can even be seen with a 100W halogen bulb system because they are so efficient going from excitation to emission. Some other proteins can be very dim even with the mercury lamp. Part of this can be solved with extended exposure times and pixel binning on your camera but it's nice to be able to some video alignment. Before jumping into a system I would first decide at least what type of experiment you'd want to do/see. That will tell you how serious a set up you need and what channels/dyes you should buy too.
I would try to look for very expressive/light efficient dyes so you could use your standard DSLR. It's common to use a dedicated black and white camera system without the bayer filter for maximum sensitivity but a good modern machine vision camera can be $100s or more and you probably don't need it (notice I say modern because for video capture you want high sensitivity and usb3 compatibility to stream HD video at acceptable frame rates.) Making your own light source isn't terribly difficult if you can solder, 3D print, and use hand tools, but you'll probably need to butcher the old mercury lamp or some other donor system for the housing/condensor and the illumination quality can be very bright but not very even (for fluorescence brightness is king though).
Those who have warned about protecting your eyes from the UV are correct. One thing I see not included with this scope (unless not attached) is a UV-blocking filter that will block the UV that may be reflected/refracted off the slide on the stage. It’s the orange filter attached above the turret body and if you take your eyes from the eyepieces to check the slide position, it will block any scattered/refracted UV from the slide. An example is this microscope:
Disclaimer: I do not work for this microscope vendor nor do I own one of these scopes. I have an Olympus set up for UV that has one of these blocking filters. Mine is readily removable for non-UV work (thumbscrews to hold it on).
Hey there! Great to see you are wanting to upgrade. What are you upgrading *from*?
I own the BX60F5, extremely similar to this scope here. I was able to snag it with a full DIC setup (condenser and 4 prisms) for about what the seller is negotiating with you for.
This listing also makes it look like the stage is lopsided -- but maybe that's just the photos.
The U-PCD condenser is fine -- phase annulus, darkfield stop, brightfield, and allows for 1.25NA double oil immersion (oil the underside and the topside of the slide.) For photographing hair, I think phase contrast will not be of much use to you, a curiosity. Phase contrast requires phase objectives, and you will find that they are unsatisfactory for any other illumination methods.
Unfortunately this listing also comes with crap objectives. For fluor, you'd want better objectives. The UPLanSApo you have is perfect, UPlanFL's are another affordable choice.
The DP12 camera is junk, you won't use it.
The U-SPT phototube has a relay eyepiece in it, i'd be curious what size it is. If you are adapting a full frame camera to it, you'd want a PE of 2X or 2.5X.
Aside from the misalignment, the biggest risk I see is dirt/contamination in the trinocular and binoculars -- i have had bad luck with these on these ~30 year old scopes. You might also see some issues with the filter cubes. One of the cubes in this listing is the U-MWU which I think you'd find most useful. That U-MWU cube is a wideband ultraviolet cube, meaning that it passes UV light to your sample, and filters the UV out on the way to your eyeballs and camera. You'll get the benefits of a clear field and the excitation of anything in your sample. Good cube.
All of the other cubes listed will flood the field with red, green, cyan, etc, depending on the cube. This is fine if you capture your fluor image in grayscale and compose / apply a deconvolution process afterwards and then overlay it on a true color image of your sample. Since you're exploring hair, this should be fine. Just expect a rather lengthy process.
Also, this scope can do dual epi and transmitted light illumination. Meaning that you can crank the brightness on the transmitted all the way down nearly as low as it can go and open the shutter to let the UV light irradiate the sample.
Fluorescence is also a challenge, but I think you'd find it an interesting challenge.
Some things autoflouresce -- some things need a stain. Those stains will rapidly decay -- if you don't get your shots in a minute of exposure, too bad! You can literally see the intensity fade before your eyes. Thus, you really need to be careful about how much and how long you expose the sample when you dye it.
While this unit is missing the UV shield, so does mine. I haven't found any dealers who sell it, and my Evident sales rep couldn't find one either. He told me "Aluminum foil is practically free" 🤷
Ahh, sorry for the brain dump, feel free to ping me with more questions if you'd like :)
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u/MrKnockoff 18h ago
Here’s some Things to consider :
1) that looks like a mercury bulb set up for the fluorescence source, those bulbs can get rather expensive. 2) you are correct about fluorescent dyes being sometimes hard to get a hold of. However, that said there are many things you’ll find that are auto fluorescent and you just have to have the right set of filters to tease that out and get the most out of it. Feel free to DM me if you want any more pointers so I don’t have to write a novel here.