Fluorescence microscopy

Images made through a microscope. All subject types.

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pwnell
Posts: 2031
Joined: Fri Dec 18, 2009 4:59 pm
Location: Tsawwassen, Canada

Fluorescence microscopy

Post by pwnell »

Depending which way I go, adding fluorescence to my microscope will cost me quite a lot of money. Before I go down this route, I'd like to get some feedback as to how much use I will have for it.

I am not a biologist, or working in a scientific lab analysing and identifying trace elements - all tasks where I believe using fluorescence is a valuable approach.

What I'd like to photograph are fluorescence in corals, possibly invertebrates and crustaceans, algae (possibly), minerals and crystals. Would I have enough subject matter to make this investment worth while? Not sure if I want to go down the route of photographing non auto-fluorescent subject matter by using fluorophores.

curt0909
Posts: 609
Joined: Wed Oct 26, 2011 10:06 am
Location: Pittsburgh, PA

Post by curt0909 »

Plants tend to autofluoresce strongly without need for dyes or special treatment. Many dyes do pose some health hazards so be careful if you go that route. I actually have 2 fluorescence microscopes, Nikon Fluophot and Reichert Diastar, but haven't used the fluorescence capability of either yet. Some day I'll get around to it.

Starshade
Posts: 133
Joined: Wed May 04, 2011 1:47 am

Post by Starshade »

I did a lot of fluorescent photography through the scope last year and I'd say it might be worth it if you like it.

But in the vast majority of cases indeed you need dyes, rubber gloves, I'd suggest a draft hood, and protective glasses.

Autofluorescence is very limited, but you certainly will find interesting objects. Keep in mind, that coral "fluorescence" is not produced with UV light. You need special filters instead and a regular lamp. I did some "fluoro" dives btw, they were awesome and certainly I didn't have any UV: just filter on my mask, and a filter on my torch. These filters are available for microscope by the same company... forgot its name, sorry. May be that will be a good start for you? It's much cheaper!

BJ
Posts: 355
Joined: Sat Sep 29, 2007 10:53 am
Location: England

Post by BJ »

Hi,

you can experiment with fluorescence microscopy quite cheaply before making a large investment.

See for example a series of articles by David Walker in Micscape magazine entitled "Forays into Fluorescence". The first article in the series outlines the approach:

http://www.microscopy-uk.org.uk/mag/art ... luor1.html

Another article on this method can be found here:

http://www.viewsfromscience.com/documen ... cence.html


Personally, as an amateur, after an intitial burst of fascination and enthusiasm, I find limited use for fluorescence.

Brian

jcb
Posts: 49
Joined: Sat Jun 11, 2011 2:38 am
Location: France

Post by jcb »

The easiest way is clearly autofluorescence. Depending on what you want to see you will need different filters.
As an example, for chlorophyll autofluorescence macrophotography (1x) I used the following :
  • excitation filter : Lee 721 berry blue or Lee 071 Tokyo blue (theater light gelatin)
    barrier filter : Heliopan R25 (light red)
You may need higher quality filters and different diameters for a microscope.

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