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So how does the manufacturer know the size of the pixel? I guess this will be calibrated at setup of the system with a target of known size for one objective and then stored somewhere in the parameters and hopfully regularly checked. Do they then just change it proportionally for the other objectives? Sometimes objective magnification can be a little bit different than what is written on the barrel, epecially when the lens has a correction collar, did anyone check this?

We had the case that a journal insisted on us giving the magnification in the figure caption, so we gave the objective magnification and insisted on having the scale bar in addition to that. 

Andreas

 

 

 

-----Original Message-----
From: Guy Cox <[log in to unmask]>
To: CONFOCALMICROSCOPY <[log in to unmask]>
Sent: Fri, 16 Aug 2013 13:01
Subject: Re: magnification


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In a confocal image the magnification depends on the objective magnification and 
the area scanned on the sample.  There is of course an 'ocular' or transfer lens 
but that is not variable and thus is included in the calibration of the system.  
So when you 'zoom' in a confocal image you are just scanning a smaller part of 
the field of view.  The system should still always give you the correct 
magnification, but since this is a digital image the appropriate units are 
pixels per micrometre (or the converse, the size of a pixel).  Every confocal 
microscope will give you these figures.  You should check once in a while, of 
course, using a standard specimen such as a stage micrometer.  

							Guy

-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On 
Behalf Of Nicola Green
Sent: Friday, 16 August 2013 8:57 PM
To: [log in to unmask]
Subject: magnification

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Hi
This is probably a very foolish and basic question, but I am going to ask
it any way and just apologise for my ignorance as a biological specialist
using a confocal microscope.

In microscopy I would generally multiply the objective lens magnification
by the ocular lens (often 10x) to get the total magnification of an image.
I want to know if this also applies for the confocal microscope or is the
light path such that the objective magnification is the only one relevant?
If I do need to include additional magnification what would these
magnifications be, are they dependent upon the system being used (LSM 510
META)  or is there a standard magnification?

Thanks for your help in clarifying this for me.

Regards
Nicola