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Author Topic: how long can it last?  (Read 7188 times)

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Offline angela

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how long can it last?
« on: February 02, 2006, 07:08:43 AM »
Hi,

I just would like to know: how long can an immunofluoerescent signal  last, according to your experience? also, what about the morphology of frozen sections with a permanent mounting?

Thanks


Angela

how long can it last?
« on: February 02, 2006, 07:08:43 AM »

Offline richard03

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how long can it last?
« Reply #1 on: February 02, 2006, 08:41:51 PM »
I have seen IF slides stored in 4 C for over a year and they are still good. It depends on several factors such as anti-fade mounting medium, the time exposed to light, storage temp, etc. Please note some fluorescent slides fade very quickly so you need to take pictures ASAP.

Mounting medium cann't help improve morphology of frozen sections. Fixation methods are important. Formaldehyde is usually giving better morphology but worsen IHC staining. Acetone/alcohol gives better IHC Staining but worsen morphology. So you have to focus on one of them for your particular application.

Richard

Offline ImmunoNYC

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how long can it last?
« Reply #2 on: February 02, 2006, 08:48:38 PM »
Perfectly said Richard.[/color]

Offline angela

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how long can it last?
« Reply #3 on: February 03, 2006, 04:16:15 AM »
Thanks, Richard and Maximina! My experience is that the fluorescence would be gone in weeks, after looking at the slides. The reason why I was asking is that I heard from somebody I work with that it can last for several years (!!!) if the slides are rinsed with toluene, and mounted with DPX.  Also, they say that the slides can be put at -20 to be stored, but I think that is not advisable.

Angela

Offline richard03

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how long can it last?
« Reply #4 on: February 03, 2006, 07:34:48 AM »
As far as I know, the anti-fading mounting medium is most important in keeping the fluorescence signal longer. But never had experience with the one (toluene, and mounted with DPX) you mentioned. I would like to hear others experience about this as well.

Richard

Offline ole

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how long can it last?
« Reply #5 on: February 03, 2006, 07:52:55 AM »
Hi

(I have no exp with toluene rinsed, dpx mounted either)
I mount with Dako fluorescense mounting medium (anti fading) . I have kept some slides in freezer (-20deg/C) for several years (educational purposes). I cant see (still) any significant loss of the fluorecent signal.
kept in frigde or roomtemp (protected from light) the signal is "stable" considerable shorter, at least in roomtemp.

Angela
The slides rinsed with toluene, and mounted with DPX? What temperature where they stored at?

Ole

Offline angela

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how long can it last?
« Reply #6 on: February 03, 2006, 08:16:16 AM »
Hi Ole,

they told me the slides were stored at -20.

Offline ole

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how long can it last?
« Reply #7 on: February 03, 2006, 08:32:25 AM »
Ok
Tnx

Offline discman

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how long can it last?
« Reply #8 on: February 03, 2006, 09:33:32 AM »
If you use water-based mounting reagents, sealing of the edges with nail polish can also improve conservation of your slides. Although some people say that the solvents in the nail polish might harm/affect the mounting medium (never observed this myself though).

I also found that 'storability' depends on the fluorophore/mounting medium combination. Although I don't remember the brand, I used a commerial mounting medium that was incompatible with the Alexa fluorophore that I used at that time (found out afterwards by reading the spec sheet...), resulting in an almost complete fading within two weeks or so.

GŁnther

how long can it last?
« Reply #8 on: February 03, 2006, 09:33:32 AM »