Nile Red dye removal

[obsydion]

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Hi all,

If anyone here is familiar with Nile Red, they would know that it is a
notoriously non-specific lipophilic dye. It especially has a penchant for
plastics and polymers, leaving a characteristic pink hue on anything it
touches. Although our lab takes care to avoid contamination by keeping 'Nile
Red free' instruments, sometimes contamination is unavoidable. Instruments
such as slice chambers, nylon hold-downs and tweezer/forceps are vulnerable
and are too expensive to throw away. I was wondering if anyone has a
protocol or even tips for cleaning and removing Nile red from instruments.
Harsh solvents are not desirable but will be considered.

Here is some information about its chemistry:
http://www.chemspider.com/Chemical-Structure.58681.html


Thank you in advance.
Kelvin

[Tamara Howard]

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The protocol for Nile blue staining of frozen sections in
Kiernan's "Histological and Histochemical Methods" bible
calls for differentiating the stain in 1% acetic acid
(aq.) - maybe that would remove dye from at least some lab
stuff as well as tissues?

Good luck!

Tamara
 
On Mon, 12 Mar 2012 13:28:27 -0500
  Kelvin Poon <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy
>listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> If anyone here is familiar with Nile Red, they would
>know that it is a
> notoriously non-specific lipophilic dye. It especially
>has a penchant for
> plastics and polymers, leaving a characteristic pink hue
>on anything it
> touches. Although our lab takes care to avoid
>contamination by keeping 'Nile
> Red free' instruments, sometimes contamination is
>unavoidable. Instruments
> such as slice chambers, nylon hold-downs and
>tweezer/forceps are vulnerable
> and are too expensive to throw away. I was wondering if
>anyone has a
> protocol or even tips for cleaning and removing Nile red
>from instruments.
> Harsh solvents are not desirable but will be considered.
>
> Here is some information about its chemistry:
> http://www.chemspider.com/Chemical-Structure.58681.html
>
>
> Thank you in advance.
> Kelvin

***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************

[David Baddeley]

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What have you tried so far? Simple things like warm water with lots of dish washing detergent might be worth a try, as might washing in cooking oil (or some other lipid/oil). I suspect that once it's actually adsorbed into a plastic though you'll have a hard time getting it out without destroying the plastic. That said, it's also unlikely to leech into anything else so it might only be a cosmetic issue.

by 2 cents,
David


________________________________
 From: Kelvin Poon <[hidden email]>
To: [hidden email]
Sent: Tuesday, 13 March 2012 7:28 AM
Subject: Nile Red dye removal
 
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To join, leave or search the confocal microscopy listserv, go to:
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Hi all,

If anyone here is familiar with Nile Red, they would know that it is a
notoriously non-specific lipophilic dye. It especially has a penchant for
plastics and polymers, leaving a characteristic pink hue on anything it
touches. Although our lab takes care to avoid contamination by keeping 'Nile
Red free' instruments, sometimes contamination is unavoidable. Instruments
such as slice chambers, nylon hold-downs and tweezer/forceps are vulnerable
and are too expensive to throw away. I was wondering if anyone has a
protocol or even tips for cleaning and removing Nile red from instruments.
Harsh solvents are not desirable but will be considered.

Here is some information about its chemistry:
http://www.chemspider.com/Chemical-Structure.58681.html


Thank you in advance.
Kelvin

[Iain Johnson]

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Nile Red physically penetrates most common laboratory plastics (e.g.
polyethylene, polystyrene), so indeed the only way of removing it
post-exposure is to dissolve the plastic.

It may be possible to mitigate the problem by pre-complexing the dye with
an anphiphilic carrier such as 2-hydroxypropyl-beta cyclodextrin,
methyl-beta cyclodextrin or BSA instead of just diluting into water from a
DMSO or ethanol stock.  For cell staining, the dye will spontaneously
transfer from the carrier to cells upon mixing.  The downside of this
approach is that Nile Red will be fluorescent in the carrier complex
whereas in water it is essentially nonfluorescent.  So you might have
additional background signals to contend with, depending on how your
labeling and imaging protocols are set up.

Biobeads SM2 (from BioRad) are a useful adsorbent for pulling hydrophobic
dyes like Nile Red out of aqueous solutions.  Unfortunately retrieval of
dye that has already penetrated exposed plastic components will be
negligible.

Iain

Iain Johnson Consulting
Eugene, OR
[phone]

On Mon, Mar 12, 2012 at 2:31 PM, David Baddeley <[hidden email]
> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> What have you tried so far? Simple things like warm water with lots of
> dish washing detergent might be worth a try, as might washing in cooking
> oil (or some other lipid/oil). I suspect that once it's actually adsorbed
> into a plastic though you'll have a hard time getting it out without
> destroying the plastic. That said, it's also unlikely to leech into
> anything else so it might only be a cosmetic issue.
>
> by 2 cents,
> David
>
>
> ________________________________
>  From: Kelvin Poon <[hidden email]>
> To: [hidden email]
> Sent: Tuesday, 13 March 2012 7:28 AM
> Subject: Nile Red dye removal
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> If anyone here is familiar with Nile Red, they would know that it is a
> notoriously non-specific lipophilic dye. It especially has a penchant for
> plastics and polymers, leaving a characteristic pink hue on anything it
> touches. Although our lab takes care to avoid contamination by keeping
> 'Nile
> Red free' instruments, sometimes contamination is unavoidable. Instruments
> such as slice chambers, nylon hold-downs and tweezer/forceps are vulnerable
> and are too expensive to throw away. I was wondering if anyone has a
> protocol or even tips for cleaning and removing Nile red from instruments.
> Harsh solvents are not desirable but will be considered.
>
> Here is some information about its chemistry:
> http://www.chemspider.com/Chemical-Structure.58681.html
>
>
> Thank you in advance.
> Kelvin
>