Re: Size of a bleach point

Posted by xavier Sanjuan on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Size-of-a-bleach-point-tp7590808p7590815.html

*****
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 for all the feedback! I did not want this for a photobleaching
experiment, we have been using a multiphoton laser at 830 nm to heat
nanoparticles within cells, but I guess this does not affect the
calculations!

Best,

Xavi.

___________________________________

*Xavier Sanjuan*
Advanced Light Microscopy Unit

Parc de Recerca Biomèdica de Barcelona
Doctor Aiguader, 88
08003 Barcelona - Spain
Tel: + 34 93 316 0195 (ext 1195 dins el PRBB)
Fax: + 34 93 316 09 01
E-mail: [hidden email]
Web:
http://www.crg.eu/en/core/programmes-groups/advanced-light-microscopy-unit


Missatge de Cammer, Michael <[hidden email]>
del dia dv., 24 d’abr. 2020 a les 18:10:

> *****
> 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.
> *****
>
> I forgot to say that in the Plexiglas reply that this worked well as a
> model for cell cultures, but we are having an issue now (ok, in February
> and maybe again someday...) that as we focus deeper in drosophila embryos
> using a Bruker galvo miniscanner that there is a lot of scattering.  At the
> surface of a zebrafish embryo we can target membranes specifically, but
> deeper in drosophila scatters too much for fine targeting.  We have tried
> three different lenses and adjusting the collar of one empirically.  This
> is at 405 and 470 nm.  So spot size is very depth and sample dependent.
>
>
> Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
>
> NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY
> 10016
>
> [hidden email]<mailto:[hidden email]>
> http://nyulmc.org/micros  http://microscopynotes.com/
>
> Voice direct only, no text or messages:  1-914-309-3270 and 1-646-501-0567
>
>
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of SANJUAN SAMARRA, XAVIER <[hidden email]>
> Sent: Thursday, April 23, 2020 6:21:30 PM
> To: [hidden email]
> Subject: Size of a bleach point
>
> [EXTERNAL]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=yP7vq3EcHuDcKVCDZvMEH2V4DrnW01XUvq51lazvtWw&s=DntAv-VWsKnK4897yAOPY2bnxV_76AqObBoVvIDrPDU&e=
> Post images on
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=yP7vq3EcHuDcKVCDZvMEH2V4DrnW01XUvq51lazvtWw&s=8UwNlUtSrvILTUYU-v7t7-F7F1BY90K5BB8Sg6aEefQ&e=
> and include the link in your posting.
> *****
>
> Dear listers,
>
> When using the Bleach Point feature in the Leica confocal systems (I'm sure
> it also exists in the other brands), I don't know how to measure the
> surface of the bleached area. I figure out the wavelength of the laser beam
> and the NA of the lens affect the size of that area, so would it be correct
> to use the resolution formula to calculate it?  Or am I mixing up things
> here?
>
> Thank you very much,
>
> Xavi.
>
> ___________________________________
>
> *Xavier Sanjuan*
> Advanced Light Microscopy Unit
>
> Parc de Recerca Biomèdica de Barcelona
> Doctor Aiguader, 88
> 08003 Barcelona - Spain
> Tel: + 34 93 316 0195 (ext 1195 dins el PRBB)
> Fax: + 34 93 316 09 01
> E-mail: [hidden email]
> Web:
>
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.crg.eu_en_core_programmes-2Dgroups_advanced-2Dlight-2Dmicroscopy-2Dunit&d=DwIFaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=E0xNnPAQpUbDiPlC50tp7rW2nBkvV7fujQf0RknE5bU&m=yP7vq3EcHuDcKVCDZvMEH2V4DrnW01XUvq51lazvtWw&s=JdJGheTLRjXZDGcKpba4K1HsNjRbLhgZKVRxqIgzx50&e=
>