Posted by
Christophe CHAMOT on
URL: http://imagej.273.s1.nabble.com/Image-transform-tp3703413p3703421.html
Liu, Dongfang (NIH/NIAID) [E] a écrit :
> Dear Gabriel,
>
> Thank you for your help.
> Yes, the system is a miroscope. Could you tell what other optics I should consider to calibrate a image? Could you give me more detail information about this?
>
> Best regards
> df
>
>
>>----------
Hello,
The formula is :
(pixel size) = (pixel camera * binning) / (total magnification)
As an example, when I use my CoolSnap (6.45µm pixel) with a 2x2 binning,
I have a "virtual" pixel of (6.45*2) 12.9 µm. This "pixel" gather a
distance through my 100x objective of 12.9/100 = 0.129 µm.
This is the pixel size I give in Image>Properties as Gabriel said
previously.
I said "total magnification" because some people use optional lenses
(zoom or optovars). I got a 1.5x on my Leica but it decreases
dramatically the intensity of fluorescence. In the previous example,
with the optovar 1.5x, pixel size will be (6.45*2)/(100*1.5)=0.086 µm
Hope it helps.
--
CHAMOT Christophe
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