Molds on decaying tangerine

Pau · Post by **Pau** » Sat Dec 10, 2016 1:18 pm

Just testing my fluorescence equipment for autofluorescence
Excitation Royal Blue 455nm LED. 10X Leitz Pl Fluotar 10/0.30 on Zeiss Standard. Stack of 77 images

ChrisR · Post by **ChrisR** » Sat Dec 10, 2016 4:16 pm

Is this epi-lighting?
If not, I assume you wouldn't need a Fluotar?

Pau · Post by **Pau** » Sat Dec 10, 2016 4:36 pm

Yes, it's classic epifluorescence with a filter cube, I used a bandpass green emission filter because all the sample was fluorescing in orangeish smearing the image.

Fluotar is the Leitz denomination for fluorite objectives, with blue excitation any other type would also work well, UV is more demanding about objective transmission.

WalterD · Post by **WalterD** » Sun Dec 11, 2016 12:50 pm

Interesting image. Are you satisfied with use of leds on subject fluorescence microscope? I myself have been using a mercury vapor arc lamp, would be interesting to retrofit the illuminator with leds. What -do you think- are the most significant diffreneces between both (if any)?

Pau · Post by **Pau** » Sun Dec 11, 2016 3:51 pm

Hi Walter,
I'm a beginner in fluorescence microscopy and I'm at the point of testing my LED setup with real subjects to take real images. I can't compare because I have no experience with HBO burners. I've bought the filters used and I'm not sure to have the right combinations in some cases

I'm very satisfied with blue LEDs, the former picture was taken at ISO 200 0.8s at 25X magnification on sensor with a 10/0.30 objective if these data can tell anything useful for you.
The main advantages of LED vs HBO are clear: no heat, instant response, much longer life (I expect), much less if any filter degradation, less dangerous...
The disadvantages I find are the low power of UV LEDs available and, with a DIY setup like mine, the need to change the LED for changing excitation and so multi spectral images will be more difficult to obtain. I'm convinced that HBO produce more intense UV radiation.

In few days (or weeks...) I will post my setup in the forum and I plan to keep posting some of my fluorescence images.

WalterD · Post by **WalterD** » Mon Dec 12, 2016 4:59 am

Thanks for the interesting explanation, Pau. The advantages are indeed big, and diy approach is easier to carry out than with the HBO burners. Sounds like an interesting opportunity, the power of available UV leds might increase as well in the future. Looking forward to hear about your future posts!

johan · Post by **johan** » Mon Dec 12, 2016 5:33 am

Likewise, very interested in this

Jacek · Post by **Jacek** » Mon Dec 12, 2016 5:46 am

Pau, poor power LED UV 365 nm, can be strengthened. For the epi illumination is actually an understatement. When lenses to UV is pretty good. I will show you my solution for small lenses for 10X and less. Upload photos because my English is google translator

Pau · Post by **Pau** » Mon Dec 12, 2016 10:55 am

I will show you my solution for small lenses for 10X and less.

Jacek, will be nice to see it!

Jacek · Post by **Jacek** » Mon Dec 12, 2016 2:24 pm

Illuminators made from the upper lens of the condenser (Zeiss, PZO).
Execution of primitive, but it works very well

Pau · Post by **Pau** » Mon Dec 12, 2016 2:54 pm

Jacek, thanks, interesting idea!

In fact with the Zeiss epifluorescence illuminator IV (the same you have mounted on the BIOLAR) the field is not full illuminated with the 4X objective, you can see some vignette at my first sorus image at the other post.

Be aware that working this way you're more exposed to UV radiation, even high intensity short blue can damage your eyes, I hope you wear UV protection glasses.

What models of UV LEDs do you use?

Jacek · Post by **Jacek** » Mon Dec 12, 2016 3:09 pm

With me on Biolar it is good, maybe because I have a cap 1.25x.
Led 3W 365 nm.
Please show how LEDs have mounted at home and how they are changing.

carlos.uruguay · Post by **carlos.uruguay** » Mon Dec 12, 2016 7:08 pm

Nice!!!!
Thanks for the explanation

Pau · Post by **Pau** » Wed Dec 14, 2016 11:03 am

Jacek wrote:Led 3W 365 nm.
Please show how LEDs have mounted at home and how they are changing.

Could you provide the link for your 3W 365 nm LED?

I've posted my fluorescence setup at the Equipment discussions forum

Jacek · Post by **Jacek** » Wed Dec 14, 2016 11:21 am

Unfortunately, after Polish:

http://allegro.pl/dioda-mocy-power-led- ... 22213.html

Put a link to your setup because I can not find