Re: How to show something inside the cell?

Posted by Kavita Aswani-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/How-to-show-something-inside-the-cell-tp7580451p7580459.html

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I can confirm Michaels comment below.  During my research life, I was labelling bacteria with FITC, and observing uptake by cultured macrophages.  Due to the conditions used, a lot of these FITC-bugs stuck to the outside of the cell, and I used Typan blue to quench the extracellular fluorescence.  Details in http://www.ncbi.nlm.nih.gov/pubmed/11726395

TITLE: Effects of surfactant protein A and NaCl concentration on the uptake of Pseudomonas aeruginosa by THP-1 cells. Khubchandani-Aswani KR, Oberley RE, Snyder JM.

Cheers,
Kavita

Kavita Aswani, PhD
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-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of MODEL, MICHAEL
Sent: June-07-13 7:04 AM
To: [hidden email]
Subject: Re: How to show something inside the cell?

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I think trypan blue has also been used to quench external fluorescence ________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Gabriel Lapointe [[hidden email]]
Sent: Thursday, June 06, 2013 5:17 PM
To: [hidden email]
Subject: Re: How to show something inside the cell?

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Hi Yi,
In the same line of thougth as Mike, you could also look at the quencher used in qPCR. Some might be compatible with your fluorophore.



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2013/6/6 MODEL, MICHAEL <[hidden email]>

> Hi Yi - I think your solution is good. Fluorophores on the surface can
> also be selectively quenched, depending on the fluorophore - FITC by
> lowering pH, red fluorophores or those emitting in the 600s by adding
> several mg/ml of Acid Blue 9.
>
> Mike Model
>
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]]
> On Behalf Of SUBSCRIBE CONFOCALMICROSCOPY
> Sent: Thursday, June 06, 2013 4:27 PM
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> Subject: How to show something inside the cell?
>
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> Dear listers,
>
> One of our user is investigating the transportation of fluorescence
> labeled DNA oligo in cells. He would like to prove the DNA oligo has
> been transported inside the cells but not on the surface of cells by
> using the light microscopy. We suggested him to clearly label the
> membrane of cells and use the Z-stack images of confocal to look through the cells.
> I am wondering if there is any other better solution?
>
> Any suggestion on good membrane dye is appreciated as well!
>
> Thank you,
>
> Yi
>
>
>

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