Re: Best parameters for optimal slicing

[Pedro Camello]

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Dear Siv,

many thanks for your table, is more complete for others I had got.
Regarding the low structures, at the moment 500 nm is enough for me

Dear Paul,

thanks for your figure. My objective is the same, though my system is a
non-UV 1024. However, at zoom = 1 my pixel size is 410 nm!. Should I
concern about a possible error in the parameters files of the software
(Lasersharp, OS/2)?

Could please anybody with a Biorad 1024 system report me the pixel size
for  60X and zoom 1?

Dear Glenn and Martin,

I thought the zoom in the Biorad 1024 was real. Are you saying that
between 2 and 4 I´m introducing garbage? I missed that line in the manual
(if present there)

Many thanks to all of you giving me a hand.

Nice list.
--
Dr Pedro J Camello
Dpt Physiology
Faculty of Veterinary Sciences
University of Extremadura
10071 Caceres
Spain
Ph: 927257100 Extension 1321
Fax:[phone]

[Glen MacDonald-2]

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On Feb 11, 2008, at 4:01 PM, Pedro J Camello wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear Siv,
>
> many thanks for your table, is more complete for others I had got.
> Regarding the low structures, at the moment 500 nm is enough for me
>
> Dear Paul,
>
> thanks for your figure. My objective is the same, though my system  
> is a
> non-UV 1024. However, at zoom = 1 my pixel size is 410 nm!. Should I
> concern about a possible error in the parameters files of the software
> (Lasersharp, OS/2)?
>
> Could please anybody with a Biorad 1024 system report me the pixel  
> size
> for  60X and zoom 1?
>
> Dear Glenn and Martin,

Yes, the zoom is real, restricting the deflection angle of the galvos  
to a smaller field.  As I recall, galvo deflection voltage on is set  
in 8-bit increments on the 1000/1024 scanner. Zooms that are not in  
multiples of 2 will have interpolation errors causing some variation  
in positioning.  I may be wrong on the details, but couldn't find the  
old discussion thread in the archives.  Pixel size at zoom =1 for any  
lens will be affected by calibration of  scan area by the service  
engineer and variation in total mag to the detector, which is  
slightly different for each system, at least for the MRC systems.

Regards,
Glen
>
> I thought the zoom in the Biorad 1024 was real. Are you saying that
> between 2 and 4 I´m introducing garbage?
> I missed that line in the manual
> (if present there)
>
I've looked for that line myself.

> Many thanks to all of you giving me a hand.
>
> Nice list.
> --
> Dr Pedro J Camello
> Dpt Physiology
> Faculty of Veterinary Sciences
> University of Extremadura
> 10071 Caceres
> Spain
> Ph: 927257100 Extension 1321
> Fax:[phone]

[William Hatton]

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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Siv,
Would it also be possible to get a copy of that table?
Will

William J. Hatton, PhD
Res. Asst. Professor
Director of Imaging and Morphology Core,
NIH Center of Biomedical Research Excellence (COBRE)
Department of Pharmacology/318
University of Nevada School of Medicine
Reno, NV 89557-0046 USA
 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Glen MacDonald
Sent: Tuesday, February 12, 2008 2:31 PM
To: [hidden email]
Subject: Re: Best parameters for optimal slicing

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

On Feb 11, 2008, at 4:01 PM, Pedro J Camello wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear Siv,
>
> many thanks for your table, is more complete for others I had got.
> Regarding the low structures, at the moment 500 nm is enough for me
>
> Dear Paul,
>
> thanks for your figure. My objective is the same, though my system  
> is a
> non-UV 1024. However, at zoom = 1 my pixel size is 410 nm!. Should I
> concern about a possible error in the parameters files of the software
> (Lasersharp, OS/2)?
>
> Could please anybody with a Biorad 1024 system report me the pixel  
> size
> for  60X and zoom 1?
>
> Dear Glenn and Martin,

Yes, the zoom is real, restricting the deflection angle of the galvos  
to a smaller field.  As I recall, galvo deflection voltage on is set  
in 8-bit increments on the 1000/1024 scanner. Zooms that are not in  
multiples of 2 will have interpolation errors causing some variation  
in positioning.  I may be wrong on the details, but couldn't find the  
old discussion thread in the archives.  Pixel size at zoom =1 for any  
lens will be affected by calibration of  scan area by the service  
engineer and variation in total mag to the detector, which is  
slightly different for each system, at least for the MRC systems.

Regards,
Glen
>
> I thought the zoom in the Biorad 1024 was real. Are you saying that
> between 2 and 4 I´m introducing garbage?
> I missed that line in the manual
> (if present there)
>
I've looked for that line myself.

> Many thanks to all of you giving me a hand.
>
> Nice list.
> --
> Dr Pedro J Camello
> Dpt Physiology
> Faculty of Veterinary Sciences
> University of Extremadura
> 10071 Caceres
> Spain
> Ph: 927257100 Extension 1321
> Fax:[phone]