For a hobbiest is there much of interest in Fluorescence?

[Gatorengineer64]

I have added an Axioscope with the upper lamp assembly and some fluorescence cubes of unknown type. Is this an avenue that many home hobbiest go down, or is this more for specialty medical? If so whats the easiest way to put your toe in the water?

[Greg Howald]

The most difficult thing about fluorescence where I am is the obtaining of fluorescent dye. Here fluorescence must be a professional thing because you must have a license to purchase it. So,,, I am leaving it alone as I am not aware of a source for this. I've googled it more than once without success. Amazon has a book on fluorescence microscopy which is a really good book at $350.00. I'd buy the book and the scope of the dye was readily available to me. If it is available it's not very popular because I can't find it in the USA.
Good luck.
Greg

[Tom Jones]

Autofluorescence. No dyes necessary. You use the intrinsic fluorescence of the organism.

https://www.microscopyu.com/references/autofluorescence

https://www.nikonsmallworld.com/galleri ... rine-algae

https://www.nikonsmallworld.com/galleri ... sp.-pollen

https://www.nikonsmallworld.com/galleri ... ing-spores

[Alexander]

Most of the dyes used to stain plant cuts produce significant fluorescence if excited by blue light. A 50W halogen lamp is enough for that.

Fluorescence stain are considered dangerous because they couple to DNA with makes them suspicious to promote cancer. On the other hand a careful person doesn't drink the stuff or rub it into their eyes.

Ebay may not be the best source for dyes but I easily found sellers for acridine orange, auramine, rhodamine b and 6G in the US or shipping into the US.
It might be a better source to ask the department of biology at the local university. You don't need a lot of stain. 4 ounces 1:10.000 solution of acridine orange in buffered water will last your whole life. All the stains I mentioned are excited by blue or green light. No need for uv lightning.

[MicroBob]

Fluorescence is beautiful for plant sections but doesn't really bring out new information in my very limited experience.
I attatch images of thick hand sections, stained with acriflavine (Part of Wacker W3A stain) and illuminated with blue light. The other picture shows autofluorescence of different mountants, not all are usable for fluorescence. For really good images one also has to control bleeding of stain into the mountant (happend in my images).

Acridine orange can be used for plancton as well and ane can get impressive results. So for technically minded microscopists who like to play with less common methods it is very attractive.

[Greg Howald]

Thank you for your information. You just made this possibility more interesting and intriguing to me.
Greg

[Gatorengineer64]

Microbob - most impressive results, I assume you are using traditional illumination with a colored filter, and not Epi illumination?

[MicroBob]

This was a quick try with a 1960s Phomi 1 and 80s III RS epi fluorescence condenser with good matched interference filters.
But transmitted fluorescence works too, just not as universally with good image quality. Lamp was Cree XHP 50 warm white (not ideal) in DIY side pipe illuminator.

[Alexander]

Just a quick and dirty sample for microscope fluorescence. This picture is taken with blue excitation from with epi-illumination. It shows some cut through a tuja wipcord needle. Picture out of camera, just reduced in size for the forum.

Flourescence_sample.jpg (109.73 KiB) Viewed 3449 times

[Hobbyst46]

The question about "interest" is really not limited to fluorescence. You might as well ask if phase contrast or brightfield or high-magnification microscopy are of interest. It really depends on one's preference. The difference between hobby and profession is not in what equipment or methodology or illumination modality one uses, but in his obligation. A hobbyist may do whatever he wishes and likes, no obligation to anything (except for domestic harmony) .

Epi fluorescence is much much easier to achieve (provided one has the equipment) than trans-illumination fluorescence, and the results are much more rewarding. It is not inexpensive, though.

The stains, IMHO, are not a severe limitation, as pointed out by others, there is auto-fluorescence (the color of which often depends on the excitation wavelength) and some stains are available.

One "limitation" of fluorescence is the same feature that becomes an advantage for professional research: the selectivity or specifity to only a few parts of the specimen. So, if you find that interesting, you will enjoy it regardless of the popularity of the method.

@MicroBob - Bob, those are great fluorescence images !

[75RR]

.
Over on www.amateurmicrography.net they doing some striking work on fluorescence

https://www.photomacrography.net/forum/ ... 2&p=273991

https://www.photomacrography.net/forum/ ... 6&p=268567

https://www.photomacrography.net/forum/ ... 4&p=274984

[BramHuntingNematodes]

The question about "interest" is really not limited to fluorescence. You might as well ask if phase contrast or brightfield or high-magnification microscopy are of interest. It really depends on one's preference.

basically if your hobby is seeing dna lit up like a christmas tree then fluorescence is of interest to you