Tetraspeck beads analogs?

[Petro]

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Hi List.
Can anyone tell me any produces of beads analogous to Tertaspeck  beads. I
have found also Gatta-beads, is there anything else on the market?
Thanks.
Petro.

--
______________________________
Petro Khoroshyy

[Romain Laine]

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Hi Petro,
Check this paper out from Ulrike Endesfelder.
https://iopscience.iop.org/article/10.1088/1361-6463/ab0862/meta

They used and compared
gold nanoparticles, TetraSpeck microspheres, FluoSpheres and nanodiamonds

There's probably a few things interesting in there for you!

Romain


*Dr Romain F. Laine*, PhD in Biophotonics
MRC Skills Development research fellow
MRC Laboratory of Molecular Cell Biology (LMCB)
University College London
Gower Street, London, WC1E 6BT
http://www.ucl.ac.uk/lmcb/users/romain-laine


On Fri, Oct 23, 2020 at 4:10 PM Khoroshyy Petro <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi List.
> Can anyone tell me any produces of beads analogous to Tertaspeck  beads. I
> have found also Gatta-beads, is there anything else on the market?
> Thanks.
> Petro.
>
> --
> ______________________________
> Petro Khoroshyy
>

[G. Esteban Fernandez]

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I use micromod sicastar-F silica particles that withstand organic solvents
(for clearing).  They make 10 nm to 1.5 um; single colors only.
https://www.micromod.de/en/produkte-22-fluorescent_sil.html

-Esteban

On Fri, Oct 23, 2020 at 7:10 AM Khoroshyy Petro <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi List.
> Can anyone tell me any produces of beads analogous to Tertaspeck  beads. I
> have found also Gatta-beads, is there anything else on the market?
> Thanks.
> Petro.
>
> --
> ______________________________
> Petro Khoroshyy
>

[Petro]

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Thanks, I'll check them out.
Petro.

On Fri, Oct 23, 2020 at 5:35 PM G. Esteban Fernandez <
[hidden email]> wrote:

> I use micromod sicastar-F silica particles that withstand organic solvents
> (for clearing).  They make 10 nm to 1.5 um; single colors only.
> https://www.micromod.de/en/produkte-22-fluorescent_sil.html
>
> -Esteban
>
> On Fri, Oct 23, 2020 at 7:10 AM Khoroshyy Petro <[hidden email]>
> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Hi List.
>> Can anyone tell me any produces of beads analogous to Tertaspeck  beads. I
>> have found also Gatta-beads, is there anything else on the market?
>> Thanks.
>> Petro.
>>
>> --
>> ______________________________
>> Petro Khoroshyy
>>
>

--
______________________________
Petro Khoroshyy

[George McNamara]

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Hi Petro and listserv,

I love Esteban's suggestion: products including particles down to 10 nm!!!

Not as small, and a lot more fluorophores, are Spherotech's FPMAS
"9plex" beads (9 fluorophores, some FRET between, 3 um diameter ... can
use low NA objective lens if want to emulate super-res stuff). Also have
other products with fewer plex.

https://www.spherotech.com/flu_par_sli.htm

FPMAS-30M9

Fluorescent PMMA Particle Slide, UV, Light Yellow, Yellow, Nile Red,
Pink, Purple, CyBlue, Sky Blue and Jade Green

https://www.spherotech.com/2020%20Product%20Detail%20Pages/Spherotech%20Fluorescent%20Particle%20Slides.pdf

Spectra for 8 shown at

https://www.spherotech.com/fluorescent%20particles%20catalog%202010-2011%20rev%20a.pdf

(9th -- Jade Green -- is in a pdf somewhere on the Spoherotech web site).

happy (and bright and well resolved) 2020,

George


On 10/23/2020 10:08 AM, Khoroshyy Petro wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi List.
> Can anyone tell me any produces of beads analogous to Tertaspeck  beads. I
> have found also Gatta-beads, is there anything else on the market?
> Thanks.
> Petro.
>

[Petro]

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Hi Romain,
Thanks a lot.

On Sun, Oct 25, 2020 at 12:54 PM Romain Laine <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Petro,
> Check this paper out from Ulrike Endesfelder.
> https://iopscience.iop.org/article/10.1088/1361-6463/ab0862/meta
>
> They used and compared
> gold nanoparticles, TetraSpeck microspheres, FluoSpheres and nanodiamonds
>
> There's probably a few things interesting in there for you!
>
> Romain
>
>
> *Dr Romain F. Laine*, PhD in Biophotonics
> MRC Skills Development research fellow
> MRC Laboratory of Molecular Cell Biology (LMCB)
> University College London
> Gower Street, London, WC1E 6BT
> http://www.ucl.ac.uk/lmcb/users/romain-laine
>
>
> On Fri, Oct 23, 2020 at 4:10 PM Khoroshyy Petro <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi List.
> > Can anyone tell me any produces of beads analogous to Tertaspeck  beads.
> I
> > have found also Gatta-beads, is there anything else on the market?
> > Thanks.
> > Petro.
> >
> > --
> > ______________________________
> > Petro Khoroshyy
> >
>

--
______________________________
Petro Khoroshyy

[Petro]

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Hi George.
Yes, the number of provided sizes is impressive in Esteban's suggestion.
Unfortunately white particles they provide only as a sample on a glass.
I would prefer some solution.
Best.
Petro.


On Sat, Oct 24, 2020 at 6:43 PM George McNamara <[hidden email]>
wrote:

> Hi Petro and listserv,
>
> I love Esteban's suggestion: products including particles down to 10 nm!!!
>
> Not as small, and a lot more fluorophores, are Spherotech's FPMAS "9plex"
> beads (9 fluorophores, some FRET between, 3 um diameter ... can use low NA
> objective lens if want to emulate super-res stuff). Also have other
> products with fewer plex.
>
> https://www.spherotech.com/flu_par_sli.htm
>
> FPMAS-30M9
>
> Fluorescent PMMA Particle Slide, UV, Light Yellow, Yellow, Nile Red, Pink,
> Purple, CyBlue, Sky Blue and Jade Green
>
>
> https://www.spherotech.com/2020%20Product%20Detail%20Pages/Spherotech%20Fluorescent%20Particle%20Slides.pdf
>
> Spectra for 8 shown at
>
>
> https://www.spherotech.com/fluorescent%20particles%20catalog%202010-2011%20rev%20a.pdf
>
> (9th -- Jade Green -- is in a pdf somewhere on the Spoherotech web site).
>
> happy (and bright and well resolved) 2020,
>
> George
>
>
> On 10/23/2020 10:08 AM, Khoroshyy Petro wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi List.
> Can anyone tell me any produces of beads analogous to Tertaspeck  beads. I
> have found also Gatta-beads, is there anything else on the market?
> Thanks.
> Petro.
>
>
>

--
______________________________
Petro Khoroshyy

[George McNamara]

*****
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*****

Hi Petro,

Contact Spherotech directly and ask to be able to buy FPMAS in
suspension (and what you consider optimal size of the particles). Almost
all of Spherotech's products are nanoparticles in suspension for flow
cytometry.

happy 2020,

George


On 10/25/2020 8:05 AM, Khoroshyy Petro wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi George.
> Yes, the number of provided sizes is impressive in Esteban's suggestion.
> Unfortunately white particles they provide only as a sample on a glass.
> I would prefer some solution.
> Best.
> Petro.
>
>
> On Sat, Oct 24, 2020 at 6:43 PM George McNamara <[hidden email]>
> wrote:
>
>> Hi Petro and listserv,
>>
>> I love Esteban's suggestion: products including particles down to 10 nm!!!
>>
>> Not as small, and a lot more fluorophores, are Spherotech's FPMAS "9plex"
>> beads (9 fluorophores, some FRET between, 3 um diameter ... can use low NA
>> objective lens if want to emulate super-res stuff). Also have other
>> products with fewer plex.
>>
>> https://www.spherotech.com/flu_par_sli.htm
>>
>> FPMAS-30M9
>>
>> Fluorescent PMMA Particle Slide, UV, Light Yellow, Yellow, Nile Red, Pink,
>> Purple, CyBlue, Sky Blue and Jade Green
>>
>>
>> https://www.spherotech.com/2020%20Product%20Detail%20Pages/Spherotech%20Fluorescent%20Particle%20Slides.pdf
>>
>> Spectra for 8 shown at
>>
>>
>> https://www.spherotech.com/fluorescent%20particles%20catalog%202010-2011%20rev%20a.pdf
>>
>> (9th -- Jade Green -- is in a pdf somewhere on the Spoherotech web site).
>>
>> happy (and bright and well resolved) 2020,
>>
>> George
>>
>>
>> On 10/23/2020 10:08 AM, Khoroshyy Petro wrote:
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Hi List.
>> Can anyone tell me any produces of beads analogous to Tertaspeck  beads. I
>> have found also Gatta-beads, is there anything else on the market?
>> Thanks.
>> Petro.
>>
>>
>>

[Zdenek Svindrych-2]

*****
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*****

From the long list of Sperotech products here:
https://www.spherotech.com/2020-2021%20Price%20List.pdf
exactly two products seem like a good alternative tot TetraSpeck beads:

FP-0257-2 Fluorescent Particles, Multiple Fluorophore, 0.2%w/v, 0.1-0.3µm
URFP-02-2 Ultra Rainbow Fluorescent Particles, ~1E10/mL, 0.1-0.3µm

Has anyone tried to look at any of these under a microscope? Small
TetraSpeck beads (0.2 um) are quite dim and it would be great to have a
brighter alternative.
I do have a good source of very bright 0.3 um beads (my favorite dry erase
markers), but these are single color only...

Best, zdenek

On Sun, Oct 25, 2020 at 8:37 AM George McNamara <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Petro,
>
> Contact Spherotech directly and ask to be able to buy FPMAS in
> suspension (and what you consider optimal size of the particles). Almost
> all of Spherotech's products are nanoparticles in suspension for flow
> cytometry.
>
> happy 2020,
>
> George
>
>
> On 10/25/2020 8:05 AM, Khoroshyy Petro wrote:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi George.
> > Yes, the number of provided sizes is impressive in Esteban's suggestion.
> > Unfortunately white particles they provide only as a sample on a glass.
> > I would prefer some solution.
> > Best.
> > Petro.
> >
> >
> > On Sat, Oct 24, 2020 at 6:43 PM George McNamara <
> [hidden email]>
> > wrote:
> >
> >> Hi Petro and listserv,
> >>
> >> I love Esteban's suggestion: products including particles down to 10
> nm!!!
> >>
> >> Not as small, and a lot more fluorophores, are Spherotech's FPMAS
> "9plex"
> >> beads (9 fluorophores, some FRET between, 3 um diameter ... can use low
> NA
> >> objective lens if want to emulate super-res stuff). Also have other
> >> products with fewer plex.
> >>
> >> https://www.spherotech.com/flu_par_sli.htm
> >>
> >> FPMAS-30M9
> >>
> >> Fluorescent PMMA Particle Slide, UV, Light Yellow, Yellow, Nile Red,
> Pink,
> >> Purple, CyBlue, Sky Blue and Jade Green
> >>
> >>
> >>
> https://www.spherotech.com/2020%20Product%20Detail%20Pages/Spherotech%20Fluorescent%20Particle%20Slides.pdf
> >>
> >> Spectra for 8 shown at
> >>
> >>
> >>
> https://www.spherotech.com/fluorescent%20particles%20catalog%202010-2011%20rev%20a.pdf
> >>
> >> (9th -- Jade Green -- is in a pdf somewhere on the Spoherotech web
> site).
> >>
> >> happy (and bright and well resolved) 2020,
> >>
> >> George
> >>
> >>
> >> On 10/23/2020 10:08 AM, Khoroshyy Petro wrote:
> >>
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >> Post images on http://www.imgur.com and include the link in your
> posting.
> >> *****
> >>
> >> Hi List.
> >> Can anyone tell me any produces of beads analogous to Tertaspeck
> beads. I
> >> have found also Gatta-beads, is there anything else on the market?
> >> Thanks.
> >> Petro.
> >>
> >>
> >>
>

--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Petro]

*****
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*****

Hi Zdenek.
Can you share more info about dry erase markers?
Thanks.
Petro.

On Sun, Oct 25, 2020, 3:14 PM Zdenek Svindrych <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> From the long list of Sperotech products here:
> https://www.spherotech.com/2020-2021%20Price%20List.pdf
> exactly two products seem like a good alternative tot TetraSpeck beads:
>
> FP-0257-2 Fluorescent Particles, Multiple Fluorophore, 0.2%w/v, 0.1-0.3µm
> URFP-02-2 Ultra Rainbow Fluorescent Particles, ~1E10/mL, 0.1-0.3µm
>
> Has anyone tried to look at any of these under a microscope? Small
> TetraSpeck beads (0.2 um) are quite dim and it would be great to have a
> brighter alternative.
> I do have a good source of very bright 0.3 um beads (my favorite dry erase
> markers), but these are single color only...
>
> Best, zdenek
>
> On Sun, Oct 25, 2020 at 8:37 AM George McNamara <[hidden email]
> >
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi Petro,
> >
> > Contact Spherotech directly and ask to be able to buy FPMAS in
> > suspension (and what you consider optimal size of the particles). Almost
> > all of Spherotech's products are nanoparticles in suspension for flow
> > cytometry.
> >
> > happy 2020,
> >
> > George
> >
> >
> > On 10/25/2020 8:05 AM, Khoroshyy Petro wrote:
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Hi George.
> > > Yes, the number of provided sizes is impressive in Esteban's
> suggestion.
> > > Unfortunately white particles they provide only as a sample on a glass.
> > > I would prefer some solution.
> > > Best.
> > > Petro.
> > >
> > >
> > > On Sat, Oct 24, 2020 at 6:43 PM George McNamara <
> > [hidden email]>
> > > wrote:
> > >
> > >> Hi Petro and listserv,
> > >>
> > >> I love Esteban's suggestion: products including particles down to 10
> > nm!!!
> > >>
> > >> Not as small, and a lot more fluorophores, are Spherotech's FPMAS
> > "9plex"
> > >> beads (9 fluorophores, some FRET between, 3 um diameter ... can use
> low
> > NA
> > >> objective lens if want to emulate super-res stuff). Also have other
> > >> products with fewer plex.
> > >>
> > >> https://www.spherotech.com/flu_par_sli.htm
> > >>
> > >> FPMAS-30M9
> > >>
> > >> Fluorescent PMMA Particle Slide, UV, Light Yellow, Yellow, Nile Red,
> > Pink,
> > >> Purple, CyBlue, Sky Blue and Jade Green
> > >>
> > >>
> > >>
> >
> https://www.spherotech.com/2020%20Product%20Detail%20Pages/Spherotech%20Fluorescent%20Particle%20Slides.pdf
> > >>
> > >> Spectra for 8 shown at
> > >>
> > >>
> > >>
> >
> https://www.spherotech.com/fluorescent%20particles%20catalog%202010-2011%20rev%20a.pdf
> > >>
> > >> (9th -- Jade Green -- is in a pdf somewhere on the Spoherotech web
> > site).
> > >>
> > >> happy (and bright and well resolved) 2020,
> > >>
> > >> George
> > >>
> > >>
> > >> On 10/23/2020 10:08 AM, Khoroshyy Petro wrote:
> > >>
> > >> *****
> > >> To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > >> Post images on http://www.imgur.com and include the link in your
> > posting.
> > >> *****
> > >>
> > >> Hi List.
> > >> Can anyone tell me any produces of beads analogous to Tertaspeck
> > beads. I
> > >> have found also Gatta-beads, is there anything else on the market?
> > >> Thanks.
> > >> Petro.
> > >>
> > >>
> > >>
> >
>
>
> --
> --
> Zdenek Svindrych, Ph.D.
> Research Scientist - Microscopy Imaging Specialist
> Department of Biochemistry and Cell Biology
> Geisel School of Medicine at Dartmouth
>

[Zdenek Svindrych-2]

*****
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*****

Hi Petro,

You can grab a marker, draw a thin line on a coverslip, and look at it
under the microscope.

If you want to get really scientific, here's what I've tried:
To get rid of any glue and possibly free dye I resuspended some beads in
IPA (isopropanol) - you don't need to cut the marker open, just dip the tip
in a 1.5mL eppendorf tube filled with IPA.
Wash several times with IPA or ethanol (spin at 14000 g for a few minutes,
discard the supernatant and resuspend in new solvent by vortexing or
sonicating).
If you suspect a lot of aggregates, you can spin shorter and keep the
supernatant.
You can then use the alcohol prep, or spin and resuspend the pellet in
water. To make the beads stick to glass, add a little bit of divalent metal
salt to the water prep (like 1 mM MgCl2).
Have fun!

The beads are chemically quite stable, I've tried mounting them in aqueous
stuff, as well as LOCA (the UV-curable glue for fixing smartphones), or
Sylgard (two-part silicone, RI=1.42, usable with autofocus systems).
The size is probably slightly below 300 nm and they are very bright.

A crop of a z-stack with a 60x lens can be found here (sparse sample):
https://drive.google.com/file/d/1DmICU1tRuLXINoOmLSgEOFj-4E5UQY50
(quite cool how Google preview copes with 16-bit multi-plane tiffs)

I almost forgot the top secret ingredient: Expo Neon dry erase markers (
https://www.uline.com/Product/Detail/S-22783/ ). Fun fact, the blue marker
fluoresces in the far red!
I only wish they made a "white" variant as well...

Best, zdenek


On Sun, Nov 1, 2020 at 5:12 PM Khoroshyy Petro <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Zdenek.
> Can you share more info about dry erase markers?
> Thanks.
> Petro.
>
> On Sun, Oct 25, 2020, 3:14 PM Zdenek Svindrych <[hidden email]> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > From the long list of Sperotech products here:
> > https://www.spherotech.com/2020-2021%20Price%20List.pdf
> > exactly two products seem like a good alternative tot TetraSpeck beads:
> >
> > FP-0257-2 Fluorescent Particles, Multiple Fluorophore, 0.2%w/v, 0.1-0.3µm
> > URFP-02-2 Ultra Rainbow Fluorescent Particles, ~1E10/mL, 0.1-0.3µm
> >
> > Has anyone tried to look at any of these under a microscope? Small
> > TetraSpeck beads (0.2 um) are quite dim and it would be great to have a
> > brighter alternative.
> > I do have a good source of very bright 0.3 um beads (my favorite dry
> erase
> > markers), but these are single color only...
> >
> > Best, zdenek
> >
> > On Sun, Oct 25, 2020 at 8:37 AM George McNamara <
> [hidden email]
> > >
> > wrote:
> >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Hi Petro,
> > >
> > > Contact Spherotech directly and ask to be able to buy FPMAS in
> > > suspension (and what you consider optimal size of the particles).
> Almost
> > > all of Spherotech's products are nanoparticles in suspension for flow
> > > cytometry.
> > >
> > > happy 2020,
> > >
> > > George
> > >
> > >
> > > On 10/25/2020 8:05 AM, Khoroshyy Petro wrote:
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > > Post images on http://www.imgur.com and include the link in your
> > > posting.
> > > > *****
> > > >
> > > > Hi George.
> > > > Yes, the number of provided sizes is impressive in Esteban's
> > suggestion.
> > > > Unfortunately white particles they provide only as a sample on a
> glass.
> > > > I would prefer some solution.
> > > > Best.
> > > > Petro.
> > > >
> > > >
> > > > On Sat, Oct 24, 2020 at 6:43 PM George McNamara <
> > > [hidden email]>
> > > > wrote:
> > > >
> > > >> Hi Petro and listserv,
> > > >>
> > > >> I love Esteban's suggestion: products including particles down to 10
> > > nm!!!
> > > >>
> > > >> Not as small, and a lot more fluorophores, are Spherotech's FPMAS
> > > "9plex"
> > > >> beads (9 fluorophores, some FRET between, 3 um diameter ... can use
> > low
> > > NA
> > > >> objective lens if want to emulate super-res stuff). Also have other
> > > >> products with fewer plex.
> > > >>
> > > >> https://www.spherotech.com/flu_par_sli.htm
> > > >>
> > > >> FPMAS-30M9
> > > >>
> > > >> Fluorescent PMMA Particle Slide, UV, Light Yellow, Yellow, Nile Red,
> > > Pink,
> > > >> Purple, CyBlue, Sky Blue and Jade Green
> > > >>
> > > >>
> > > >>
> > >
> >
> https://www.spherotech.com/2020%20Product%20Detail%20Pages/Spherotech%20Fluorescent%20Particle%20Slides.pdf
> > > >>
> > > >> Spectra for 8 shown at
> > > >>
> > > >>
> > > >>
> > >
> >
> https://www.spherotech.com/fluorescent%20particles%20catalog%202010-2011%20rev%20a.pdf
> > > >>
> > > >> (9th -- Jade Green -- is in a pdf somewhere on the Spoherotech web
> > > site).
> > > >>
> > > >> happy (and bright and well resolved) 2020,
> > > >>
> > > >> George
> > > >>
> > > >>
> > > >> On 10/23/2020 10:08 AM, Khoroshyy Petro wrote:
> > > >>
> > > >> *****
> > > >> To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > >> Post images on http://www.imgur.com and include the link in your
> > > posting.
> > > >> *****
> > > >>
> > > >> Hi List.
> > > >> Can anyone tell me any produces of beads analogous to Tertaspeck
> > > beads. I
> > > >> have found also Gatta-beads, is there anything else on the market?
> > > >> Thanks.
> > > >> Petro.
> > > >>
> > > >>
> > > >>
> > >
> >
> >
> > --
> > --
> > Zdenek Svindrych, Ph.D.
> > Research Scientist - Microscopy Imaging Specialist
> > Department of Biochemistry and Cell Biology
> > Geisel School of Medicine at Dartmouth
> >
>

--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Petro]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Thanks, Zdenek.
I will have a look where I can order them, and try some which I am kicking
around.
Best.
Petro.

On Mon, Nov 2, 2020 at 4:34 AM Zdenek Svindrych <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Petro,
>
> You can grab a marker, draw a thin line on a coverslip, and look at it
> under the microscope.
>
> If you want to get really scientific, here's what I've tried:
> To get rid of any glue and possibly free dye I resuspended some beads in
> IPA (isopropanol) - you don't need to cut the marker open, just dip the tip
> in a 1.5mL eppendorf tube filled with IPA.
> Wash several times with IPA or ethanol (spin at 14000 g for a few minutes,
> discard the supernatant and resuspend in new solvent by vortexing or
> sonicating).
> If you suspect a lot of aggregates, you can spin shorter and keep the
> supernatant.
> You can then use the alcohol prep, or spin and resuspend the pellet in
> water. To make the beads stick to glass, add a little bit of divalent metal
> salt to the water prep (like 1 mM MgCl2).
> Have fun!
>
> The beads are chemically quite stable, I've tried mounting them in aqueous
> stuff, as well as LOCA (the UV-curable glue for fixing smartphones), or
> Sylgard (two-part silicone, RI=1.42, usable with autofocus systems).
> The size is probably slightly below 300 nm and they are very bright.
>
> A crop of a z-stack with a 60x lens can be found here (sparse sample):
> https://drive.google.com/file/d/1DmICU1tRuLXINoOmLSgEOFj-4E5UQY50
> (quite cool how Google preview copes with 16-bit multi-plane tiffs)
>
> I almost forgot the top secret ingredient: Expo Neon dry erase markers (
> https://www.uline.com/Product/Detail/S-22783/ ). Fun fact, the blue marker
> fluoresces in the far red!
> I only wish they made a "white" variant as well...
>
> Best, zdenek
>
>
> On Sun, Nov 1, 2020 at 5:12 PM Khoroshyy Petro <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi Zdenek.
> > Can you share more info about dry erase markers?
> > Thanks.
> > Petro.
> >
> > On Sun, Oct 25, 2020, 3:14 PM Zdenek Svindrych <[hidden email]>
> wrote:
> >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > From the long list of Sperotech products here:
> > > https://www.spherotech.com/2020-2021%20Price%20List.pdf
> > > exactly two products seem like a good alternative tot TetraSpeck beads:
> > >
> > > FP-0257-2 Fluorescent Particles, Multiple Fluorophore, 0.2%w/v,
> 0.1-0.3µm
> > > URFP-02-2 Ultra Rainbow Fluorescent Particles, ~1E10/mL, 0.1-0.3µm
> > >
> > > Has anyone tried to look at any of these under a microscope? Small
> > > TetraSpeck beads (0.2 um) are quite dim and it would be great to have a
> > > brighter alternative.
> > > I do have a good source of very bright 0.3 um beads (my favorite dry
> > erase
> > > markers), but these are single color only...
> > >
> > > Best, zdenek
> > >
> > > On Sun, Oct 25, 2020 at 8:37 AM George McNamara <
> > [hidden email]
> > > >
> > > wrote:
> > >
> > > > *****
> > > > To join, leave or search the confocal microscopy listserv, go to:
> > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > > Post images on http://www.imgur.com and include the link in your
> > > posting.
> > > > *****
> > > >
> > > > Hi Petro,
> > > >
> > > > Contact Spherotech directly and ask to be able to buy FPMAS in
> > > > suspension (and what you consider optimal size of the particles).
> > Almost
> > > > all of Spherotech's products are nanoparticles in suspension for flow
> > > > cytometry.
> > > >
> > > > happy 2020,
> > > >
> > > > George
> > > >
> > > >
> > > > On 10/25/2020 8:05 AM, Khoroshyy Petro wrote:
> > > > > *****
> > > > > To join, leave or search the confocal microscopy listserv, go to:
> > > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > > > Post images on http://www.imgur.com and include the link in your
> > > > posting.
> > > > > *****
> > > > >
> > > > > Hi George.
> > > > > Yes, the number of provided sizes is impressive in Esteban's
> > > suggestion.
> > > > > Unfortunately white particles they provide only as a sample on a
> > glass.
> > > > > I would prefer some solution.
> > > > > Best.
> > > > > Petro.
> > > > >
> > > > >
> > > > > On Sat, Oct 24, 2020 at 6:43 PM George McNamara <
> > > > [hidden email]>
> > > > > wrote:
> > > > >
> > > > >> Hi Petro and listserv,
> > > > >>
> > > > >> I love Esteban's suggestion: products including particles down to
> 10
> > > > nm!!!
> > > > >>
> > > > >> Not as small, and a lot more fluorophores, are Spherotech's FPMAS
> > > > "9plex"
> > > > >> beads (9 fluorophores, some FRET between, 3 um diameter ... can
> use
> > > low
> > > > NA
> > > > >> objective lens if want to emulate super-res stuff). Also have
> other
> > > > >> products with fewer plex.
> > > > >>
> > > > >> https://www.spherotech.com/flu_par_sli.htm
> > > > >>
> > > > >> FPMAS-30M9
> > > > >>
> > > > >> Fluorescent PMMA Particle Slide, UV, Light Yellow, Yellow, Nile
> Red,
> > > > Pink,
> > > > >> Purple, CyBlue, Sky Blue and Jade Green
> > > > >>
> > > > >>
> > > > >>
> > > >
> > >
> >
> https://www.spherotech.com/2020%20Product%20Detail%20Pages/Spherotech%20Fluorescent%20Particle%20Slides.pdf
> > > > >>
> > > > >> Spectra for 8 shown at
> > > > >>
> > > > >>
> > > > >>
> > > >
> > >
> >
> https://www.spherotech.com/fluorescent%20particles%20catalog%202010-2011%20rev%20a.pdf
> > > > >>
> > > > >> (9th -- Jade Green -- is in a pdf somewhere on the Spoherotech web
> > > > site).
> > > > >>
> > > > >> happy (and bright and well resolved) 2020,
> > > > >>
> > > > >> George
> > > > >>
> > > > >>
> > > > >> On 10/23/2020 10:08 AM, Khoroshyy Petro wrote:
> > > > >>
> > > > >> *****
> > > > >> To join, leave or search the confocal microscopy listserv, go to:
> > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > > >> Post images on http://www.imgur.com and include the link in your
> > > > posting.
> > > > >> *****
> > > > >>
> > > > >> Hi List.
> > > > >> Can anyone tell me any produces of beads analogous to Tertaspeck
> > > > beads. I
> > > > >> have found also Gatta-beads, is there anything else on the market?
> > > > >> Thanks.
> > > > >> Petro.
> > > > >>
> > > > >>
> > > > >>
> > > >
> > >
> > >
> > > --
> > > --
> > > Zdenek Svindrych, Ph.D.
> > > Research Scientist - Microscopy Imaging Specialist
> > > Department of Biochemistry and Cell Biology
> > > Geisel School of Medicine at Dartmouth
> > >
> >
>
>
> --
> --
> Zdenek Svindrych, Ph.D.
> Research Scientist - Microscopy Imaging Specialist
> Department of Biochemistry and Cell Biology
> Geisel School of Medicine at Dartmouth
>

--
______________________________
Petro Khoroshyy

[Christian Feldhaus]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi,

if you just want to measure PSFs and don't necessarily need all colours
in one bead you can take highlighter pens to create a sample. Many types
of pens work for beads with 300nm diameter or more. If you want to be on
the subresolution side (e.g. to measure high NA objectives) the only one
suitable we found so far are the Zebra Mildliner pastel ones (Zebra Pen
product number 78105). We had a poster on this at FOM 2019 and a
workshop at TIM 2020. You can find all currently available information
on this project (including sample prep) here:

http://webdav.tuebingen.mpg.de/LM/FOM2019/

If you have further questions please let me know.

Best,

Christian


On 02.11.20 09:35, Khoroshyy Petro wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Thanks, Zdenek.
> I will have a look where I can order them, and try some which I am kicking
> around.
> Best.
> Petro.
>
> On Mon, Nov 2, 2020 at 4:34 AM Zdenek Svindrych <[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Hi Petro,
>>
>> You can grab a marker, draw a thin line on a coverslip, and look at it
>> under the microscope.
>>
>> If you want to get really scientific, here's what I've tried:
>> To get rid of any glue and possibly free dye I resuspended some beads in
>> IPA (isopropanol) - you don't need to cut the marker open, just dip
>> the tip
>> in a 1.5mL eppendorf tube filled with IPA.
>> Wash several times with IPA or ethanol (spin at 14000 g for a few
>> minutes,
>> discard the supernatant and resuspend in new solvent by vortexing or
>> sonicating).
>> If you suspect a lot of aggregates, you can spin shorter and keep the
>> supernatant.
>> You can then use the alcohol prep, or spin and resuspend the pellet in
>> water. To make the beads stick to glass, add a little bit of divalent
>> metal
>> salt to the water prep (like 1 mM MgCl2).
>> Have fun!
>>
>> The beads are chemically quite stable, I've tried mounting them in
>> aqueous
>> stuff, as well as LOCA (the UV-curable glue for fixing smartphones), or
>> Sylgard (two-part silicone, RI=1.42, usable with autofocus systems).
>> The size is probably slightly below 300 nm and they are very bright.
>>
>> A crop of a z-stack with a 60x lens can be found here (sparse sample):
>> https://drive.google.com/file/d/1DmICU1tRuLXINoOmLSgEOFj-4E5UQY50
>> (quite cool how Google preview copes with 16-bit multi-plane tiffs)
>>
>> I almost forgot the top secret ingredient: Expo Neon dry erase markers (
>> https://www.uline.com/Product/Detail/S-22783/ ). Fun fact, the blue
>> marker
>> fluoresces in the far red!
>> I only wish they made a "white" variant as well...
>>
>> Best, zdenek
>>
>>
>> On Sun, Nov 1, 2020 at 5:12 PM Khoroshyy Petro <[hidden email]>
>> wrote:
>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> Post images on http://www.imgur.com and include the link in your
>> posting.
>>> *****
>>>
>>> Hi Zdenek.
>>> Can you share more info about dry erase markers?
>>> Thanks.
>>> Petro.
>>>
>>> On Sun, Oct 25, 2020, 3:14 PM Zdenek Svindrych <[hidden email]>
>> wrote:
>>>> *****
>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>> Post images on http://www.imgur.com and include the link in your
>>> posting.
>>>> *****
>>>>
>>>> From the long list of Sperotech products here:
>>>> https://www.spherotech.com/2020-2021%20Price%20List.pdf
>>>> exactly two products seem like a good alternative tot TetraSpeck beads:
>>>>
>>>> FP-0257-2 Fluorescent Particles, Multiple Fluorophore, 0.2%w/v,
>> 0.1-0.3µm
>>>> URFP-02-2 Ultra Rainbow Fluorescent Particles, ~1E10/mL, 0.1-0.3µm
>>>>
>>>> Has anyone tried to look at any of these under a microscope? Small
>>>> TetraSpeck beads (0.2 um) are quite dim and it would be great to have a
>>>> brighter alternative.
>>>> I do have a good source of very bright 0.3 um beads (my favorite dry
>>> erase
>>>> markers), but these are single color only...
>>>>
>>>> Best, zdenek
>>>>
>>>> On Sun, Oct 25, 2020 at 8:37 AM George McNamara <
>>> [hidden email]
>>>> wrote:
>>>>
>>>>> *****
>>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>>> Post images on http://www.imgur.com and include the link in your
>>>> posting.
>>>>> *****
>>>>>
>>>>> Hi Petro,
>>>>>
>>>>> Contact Spherotech directly and ask to be able to buy FPMAS in
>>>>> suspension (and what you consider optimal size of the particles).
>>> Almost
>>>>> all of Spherotech's products are nanoparticles in suspension for flow
>>>>> cytometry.
>>>>>
>>>>> happy 2020,
>>>>>
>>>>> George
>>>>>
>>>>>
>>>>> On 10/25/2020 8:05 AM, Khoroshyy Petro wrote:
>>>>>> *****
>>>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>>>> Post images on http://www.imgur.com and include the link in your
>>>>> posting.
>>>>>> *****
>>>>>>
>>>>>> Hi George.
>>>>>> Yes, the number of provided sizes is impressive in Esteban's
>>>> suggestion.
>>>>>> Unfortunately white particles they provide only as a sample on a
>>> glass.
>>>>>> I would prefer some solution.
>>>>>> Best.
>>>>>> Petro.
>>>>>>
>>>>>>
>>>>>> On Sat, Oct 24, 2020 at 6:43 PM George McNamara <
>>>>> [hidden email]>
>>>>>> wrote:
>>>>>>
>>>>>>> Hi Petro and listserv,
>>>>>>>
>>>>>>> I love Esteban's suggestion: products including particles down to
>> 10
>>>>> nm!!!
>>>>>>> Not as small, and a lot more fluorophores, are Spherotech's FPMAS
>>>>> "9plex"
>>>>>>> beads (9 fluorophores, some FRET between, 3 um diameter ... can
>> use
>>>> low
>>>>> NA
>>>>>>> objective lens if want to emulate super-res stuff). Also have
>> other
>>>>>>> products with fewer plex.
>>>>>>>
>>>>>>> https://www.spherotech.com/flu_par_sli.htm
>>>>>>>
>>>>>>> FPMAS-30M9
>>>>>>>
>>>>>>> Fluorescent PMMA Particle Slide, UV, Light Yellow, Yellow, Nile
>> Red,
>>>>> Pink,
>>>>>>> Purple, CyBlue, Sky Blue and Jade Green
>>>>>>>
>>>>>>>
>>>>>>>
>> https://www.spherotech.com/2020%20Product%20Detail%20Pages/Spherotech%20Fluorescent%20Particle%20Slides.pdf
>>>>>>> Spectra for 8 shown at
>>>>>>>
>>>>>>>
>>>>>>>
>> https://www.spherotech.com/fluorescent%20particles%20catalog%202010-2011%20rev%20a.pdf
>>>>>>> (9th -- Jade Green -- is in a pdf somewhere on the Spoherotech web
>>>>> site).
>>>>>>> happy (and bright and well resolved) 2020,
>>>>>>>
>>>>>>> George
>>>>>>>
>>>>>>>
>>>>>>> On 10/23/2020 10:08 AM, Khoroshyy Petro wrote:
>>>>>>>
>>>>>>> *****
>>>>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>>>>> Post images on http://www.imgur.com and include the link in your
>>>>> posting.
>>>>>>> *****
>>>>>>>
>>>>>>> Hi List.
>>>>>>> Can anyone tell me any produces of beads analogous to Tertaspeck
>>>>> beads. I
>>>>>>> have found also Gatta-beads, is there anything else on the market?
>>>>>>> Thanks.
>>>>>>> Petro.
>>>>>>>
>>>>>>>
>>>>>>>
>>>>
>>>> --
>>>> --
>>>> Zdenek Svindrych, Ph.D.
>>>> Research Scientist - Microscopy Imaging Specialist
>>>> Department of Biochemistry and Cell Biology
>>>> Geisel School of Medicine at Dartmouth
>>>>
>>
>> --
>> --
>> Zdenek Svindrych, Ph.D.
>> Research Scientist - Microscopy Imaging Specialist
>> Department of Biochemistry and Cell Biology
>> Geisel School of Medicine at Dartmouth
>>
>

--
Christian Feldhaus, PhD
Light Microscopy Facility
MPI for Developmental Biology

Max-Planck-Ring 5 (Spemannstrasse 35)
72076 Tübingen
Germany

deliveries to:
Max-Planck-Ring 1
72076 Tübingen

Tel.: ++49 7071 601443