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Thank you, Andrea!
What a nice discussion and explanation! I appreciate the reply,
and I'll keep a copy of your e-mail handy.
Graci!
Kathy
At 04:39 PM 12/2/02 +0100, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Dear Kathy,
>the maximum resolution for optic microscopy should be 200 nm, according
>to standard physics laws: that means that any higher resolution will
>give you no more information on the sample.
>As for useful formulas:
>- the Nyquist theorem for lateral resolution should be similar to
>X= 0,4*lambda/NA, where NA is numerical aperture of your objective and
>lambda the wavelength. It means that, if you’re working with 63x
>objective and 1,32 as NA, at 488nm, the lateral resolution is 148nm;
>dividing this number by 3, you obtain almost 49 nm. 49 is the raster
>distance you should set in your scan, in order to avoid loss of info
>from the specimen —> it does NOT mean that you will obtain resolution
>higher than 200nm, but only that you’ll obtain good data by scanning
>thrice the 148nm distance obtained before; in order to modify the
>raster distance you should change the electronic zoom and the scan
>format.
>At least, this is what I've found; if anyone else has nice data, I'll
>be interested as you in reading more :)
>Thank you and have a nice time.
>AM
>
>
>
>
>dott. Andrea Manazza
>Dipartimento di Oncologia, Università di Torino
>CeRMS, ospedale Molinette
>Via Santena, 7 - 10126 Torino, Italia
>tel: +39-11-6706522
>lab: +39-11-6336859
>fax: +39-11-6336887
>
>----- Original Message -----
>From: Kathy Spencer <[log in to unmask]>
>Date: Wednesday, November 27, 2002 9:32 pm
>Subject: confocal resolution
>
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > Hello All!
> > Basic generic question....what is the limit of resolution
> > on a
> > confocal? I'm using an Olympus Fluoview confocal, 60x 1.4NA
> > objective, oil
> > (1.516), pinhole automatically set for 1 Airy disk. For 488nm
> > laser, I
> > calculate 174nm using resolution=wavelength/2NA. But can the confocal
> > resolve more than that? What is a good formula?
> > Thanks!
> > Kathy
> >
> >
> >
> >
> >
> >
> >
> >
> > Kathy Spencer
> > The Scripps Research Institute
> > 10550 N. Torrey Pines Road
> > ICND 202
> > La Jolla, CA 92037
> > 858-784-8437
> >
Kathy Spencer
The Scripps Research Institute
10550 N. Torrey Pines Road
ICND 202
La Jolla, CA 92037
858-784-8437
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