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Many thanks, everyone! I will try Nile Red/Hoechst and will let you know

Mike

-----Original Message-----
From: Confocal Microscopy List <[log in to unmask]> On Behalf Of Lorenzo Scipioni
Sent: Thursday, April 11, 2024 7:56 PM
To: [log in to unmask]
Subject: Re: EXT: Re: vesicles

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Hi everyone,

I'm pretty sure they're lipid droplets, the high contrast is due to the fact that they're almost completely fat and the lipid/water interface gives a lot of contrast.

I wouldn't rule out mycoplasma since it can be very annoying. I propose to double stain it with hoechst (positive if mycoplasma) and any of the bodipy dyes or Nile red (positive if lipid droplets) to sort it out.

Might also be lysosomes but you said they're not acidic.

Just me two cents!
Best

Lorenzo Scipioni

On Thu, Apr 11, 2024, 16:52 Model, Michael <[log in to unmask]> wrote:

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>
> Interesting! If this were so, would Hoechst stain them?
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Kilgore, Jason A.
> Sent: Thursday, April 11, 2024 7:44 PM
> To: [log in to unmask]
> Subject: Re: EXT: Re: vesicles
>
> *****
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>
> Michael,
>
> I hope I'm wrong about this, but it reminds me of images of
> mycoplasma-infected cells. Those cells also don't look very healthy.
> Could they be infected?
>
> Jason
>
> Jason A. Kilgore
> Supervisor, Cell Analysis Tech Support Life Sciences Solutions Thermo
> Fisher Scientific
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Model, Michael
> Sent: Thursday, April 11, 2024 4:38 PM
> To: [log in to unmask]
> Subject: Re: EXT: Re: vesicles
>
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>
> Thank you, Tim,
> But I don't think this is it. I placed one of the images here:
>
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> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Tim F
> Sent: Thursday, April 11, 2024 5:24 PM
> To: [log in to unmask]
> Subject: EXT: Re: vesicles
>
> *****
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>
> Hi Michael, just to confirm, if your experience is similar to the
> video below, do you mean the dark spots? Thanks to Nikon for the vid!
>
>
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>
> I remember thinking those are protein mis-folding aggregates. I don't
> think you would see endosomes with that kind of contrast, and vesicles
> are too small at 60nm to see with transmitted light.
>
> Best,
>
>
> T
>
> Timothy Feinstein, Ph.D.
>
> On Thu, Apr 11, 2024 at 3:53 PM Model, Michael <[log in to unmask]> wrote:
>
> > *****
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> > iLCJXVCI6Mn0%3D%7C0%7C%7C%7C&sdata=e6%2Fjtgn4Xq4zGhPwdOJ7CiaDaEUdiUz
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> > p5m2A0psiw%3D&reserved=0
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> > kn
> > NFn86y8bg$
> > s.umn.edu%2Fcgi-bin%2Fwa%3FSUBED1%3Dconfocalmicroscopy%26A%3D1&data=
> > 05
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> > *****
> >
> > Dear List,
> >
> > If you look at cultured cells, such as HeLa, under bright field, you
> > clearly see many vesicles about 1 um. Does anyone know what they are?
> > (They are not acidic). The stupidest thing is that I once found a
> > paper that investigates them, but now I cannot find that paper!
> >
> > Mike
> >
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