Yes, you understood it well, at the broadband BS test I didn't use excitation filter, juts the light source, the BS and the EX filter.
Now I've tested the broadband BS with the adequate Ex and Em filters and it works fine, although brightness is clearly inferior than with the matched dichroic BS as expected
Multi LED epifluorescence illuminator
Moderators: rjlittlefield, ChrisR, Chris S., Pau
Pau,
I have an idea of doing cheap fluorescence using Lee filters and immersion darkfield condenser, without using epi light. Results will be much worse, for sure. But much cheaper.
Lee filters are very cheap at about $7 shipped for 6" diameter. They offer proper spectrum data too.
For example, for chlorophyll auto fluorescence, assuming excitation of 420-460 nm, we can use Lee #132 or #068:
http://www.leefilters.com/lighting/colo ... &filter=cf
Do you remember what red emission spectrum should I use for chlorophyll auto fluorescence? I think I should be able to find a Lee red filter for cheap too.
Edit: it says > 600nm here: http://www.biotechniques.com/Biotechniq ... ?pageNum=1
Looking through Lee filter's web site, it seems that #787, #027 and #029 may be best for > 600 nm. Not exactly sure.......
I already have good darkfield and light source, so it is cheap to try the Lee filters. I will report back.
I have an idea of doing cheap fluorescence using Lee filters and immersion darkfield condenser, without using epi light. Results will be much worse, for sure. But much cheaper.
Lee filters are very cheap at about $7 shipped for 6" diameter. They offer proper spectrum data too.
For example, for chlorophyll auto fluorescence, assuming excitation of 420-460 nm, we can use Lee #132 or #068:
http://www.leefilters.com/lighting/colo ... &filter=cf
Do you remember what red emission spectrum should I use for chlorophyll auto fluorescence? I think I should be able to find a Lee red filter for cheap too.
Edit: it says > 600nm here: http://www.biotechniques.com/Biotechniq ... ?pageNum=1
Looking through Lee filter's web site, it seems that #787, #027 and #029 may be best for > 600 nm. Not exactly sure.......
I already have good darkfield and light source, so it is cheap to try the Lee filters. I will report back.
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Ichthyophthirius
- Posts: 1152
- Joined: Thu Mar 07, 2013 5:24 am
it doesn't seem fool-proof. I know, it won't zap your retina at 364 nm, but still ...Fan, I have no experience with Lee filters, but taking a quick look at the spectra of the ones you proposed some of them are very far of the desirable specification for fluorescence, this is logical being intended for a very different purpose (see for ex. the graphs a Chroma site, miscroscopyu, etc)
Doing fluorescence with transmitted dark field is a classic method nowadays abandoned for the much more efficient epi fluorescence, but well implemented will work for sure. Be aware that typically it was done with monster HBO lamps. You need an efficient reflective condenser (cardioid...) almost for sure oil immersed. Stop type condensers trow away too much light.
Anyway it will be very interesting to know how it works if you do it.
Doing fluorescence with transmitted dark field is a classic method nowadays abandoned for the much more efficient epi fluorescence, but well implemented will work for sure. Be aware that typically it was done with monster HBO lamps. You need an efficient reflective condenser (cardioid...) almost for sure oil immersed. Stop type condensers trow away too much light.
Anyway it will be very interesting to know how it works if you do it.
Pau
Ichty, thanks for your comment.
Yes, my setup would not pass any safety inspection, nor even about ergonomy but it works
It is not fool-proof , and I'm somewhat fool, too often I forget to change the slider after changing the commuter, and I'm thinking how to design some protection (shading...)
An enclosed wheel commanded by a step motor computer controlled will be much better and is doable for people with better skills...but for me this is quite an achievement
Yes, my setup would not pass any safety inspection, nor even about ergonomy but it works
It is not fool-proof , and I'm somewhat fool, too often I forget to change the slider after changing the commuter, and I'm thinking how to design some protection (shading...)
An enclosed wheel commanded by a step motor computer controlled will be much better and is doable for people with better skills...but for me this is quite an achievement
Pau