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Perhaps this paper - just published - might have an interesting nugget regarding this? (possibly could be used in fixed cells as well)
Constitutive expression of a fluorescent protein reports the size of
live human cells
Daniel F. Berenson, Evgeny Zatulovskiy, Shicong Xie, and Jan M.
Skotheim
Molecular Biology of the Cell, Vol. 0, No. 0: mbc.E19-03-0171.
https://www.molbiolcell.org/doi/10.1091/mbc.E19-03-0171?ai=25zg&ui=2rn1&af=T
On 11/10/2019, 18:31, "Confocal Microscopy List on behalf of Mike Nelson" <[log in to unmask] on behalf of [log in to unmask]> wrote:
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This reminded me of a thread on the microscopy forum where the effects of
various mounting media types on the structures of the fixed cells were
compared. Might be worth a look.
https://forum.microlist.org/t/hardening-mounting-media-that-doesnt-compress-sample/416
Mike
On Fri, Oct 11, 2019, 2:51 AM Jeremy Adler <[log in to unmask]> wrote:
> *****
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>
> You could try fluorescent microspheres - I would expect 0.5um to be
> excluded from fixed cells.
> Your fixed cells are likely to shrink in the Z axis - understating their
> volume.
>
> Jeremy Adler
> BioVis
> Uppsala U
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Sylvie Le Guyader
> Sent: den 10 oktober 2019 17:15
> To: [log in to unmask]
> Subject: Re: Measuring cell volume by negative staining
>
> *****
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>
> But I forgot to mention that our cells are fixed and permeabilized which
> makes it more complicated. We need to find a dye that is large enough to
> stay outside. :-/
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Facility Manager
> Karolinska Institutet- Bionut Dpt
> Blickagången 16,
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> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of [log in to unmask]
> Sent: 10 October 2019 16:53
> To: [log in to unmask]
> Subject: Re: Measuring cell volume by negative staining
>
> *****
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>
> Hi Sylvie,
>
> I am not sure this is the paper you are looking for, but in 2018 Valentin
> Nägerl's group published a paper on "super-resolution shadow imaging"
> (SUSHI), in which they measured the extracellular space in brain tissue.
> Very nice paper, with lots of different applications, but maybe not what
> you had in mind.
> The reference is here:
>
> https://eur01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fpubmed%2F29474910&data=02%7C01%7Csylvie.le.guyader%40KI.SE%7C6aba2a5736224f1042f808d74d91b386%7Cbff7eef1cf4b4f32be3da1dda043c05d%7C0%7C0%7C637063160488403234&sdata=IQeuqKb215p409D%2BDkCaFdQjSz7n4G2gkB%2FrcFPaYS4%3D&reserved=0
> DOI: 10.1016/j.cell.2018.02.007
>
> Best,
> Nicolai
>
> >>>>>>>>>><<<<<<<<>>>>>>>>>><<<<<<<<<<>>>>>>>>>><<<<<<<<<<
> Dr. Nicolai T. Urban
> Imaging Center @ Max Planck Florida Institute Jupiter, 33458 FL, USA
>
>
>
>
> -----Original Message-----
> From: Confocal Microscopy List <[log in to unmask]> On
> Behalf Of Sylvie Le Guyader
> Sent: Thursday, 10 October 2019 10:31
> To: [log in to unmask]
> Subject: Measuring cell volume by negative staining
>
> *****
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>
> Dear list
>
> I remember reading a paper where the authors wanted to precisely measure
> the volume of cells . They added a fluorophore to the medium, acquired a z
> stack and segmented the negative areas in the image to find the cells.
> I cannot find this paper. Would anyone have a reference?
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Facility Manager
> Karolinska Institutet- Bionut Dpt
> Blickagången 16,
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> 14157 Huddinge, Sweden
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