Re: How to show something inside the cell?

Posted by Stephen Firth (Med) on
URL: http://confocal-microscopy-list.275.s1.nabble.com/How-to-show-something-inside-the-cell-tp7580451p7580456.html

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Another possibility is using a membrane lawn, sorry can't remember the
reference. Essentially run the experiment, staining etc but shear the cell
off the coverslip leaving the membrane behind. If the target is on the
membrane it will be visible, but if it has moved through into the cytoplasm
or beyond it will have been removed.
Cheers,
Stephen


On 7 June 2013 07:17, Gabriel Lapointe <[hidden email]>wrote:

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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.
>
>
>
> *Gabriel Lapointe, M.Sc.*
> Lab Manager / Microscopy Specialist
> Concordia University, Biology Department
> 7141 Sherbrooke St. West SP 534
> Montréal QC H4B 1R6 Canada
> Lab : [phone] x5988
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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
> > To: [hidden email]
> > 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
> >
> >
> >
>

--
Stephen Firth
Manager of Advanced Optical Microscopy
Monash Micro Imaging
Monash University
Room G53A Building 75
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