Posted by
Yang, Ann-Fook on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Objective-for-confocal-tp590639p590647.html
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalHi Cameron,
Thank you very much for sharing your experience. I would appreciate receiving the report you mentioned.
Regards,
Ann Fook
-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Nowell, Cameron
Sent: Friday, October 26, 2007 9:08 PM
To:
[hidden email]
Subject: Re: Objective for confocal
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalHi Ann,
To follow up from Julio's comments. The higher NA will give you improved resolution. A 1.42 NA object theoreticatly can resolve down to 210nm at 488nm exitation. That holds true for what ever the magnification the objective has.
We have compared a 63x 1.42 NA to a 100x 1.42 NA objective. Both were Olympus UIS2 PlanApo objectives. Both could resolve the same structures. But the big difference was the amount of light that was needed to capture an image with the 100x objective. On average 7x more light input was required to get image with the 100x lens that was equivalent to the 63x lens.
So my advice woudl be to get a 63x 1.42 NA objective, or better yet a 40x 1.4NA if you can find one (they are being made by Nikon i think but not sure about others).
Of course this is only important if you want to b e able to resolve very small structures. If you do not want to be resolving down to 210nm then your 40x oil and 63x water imersion objective will be just fine.
I can send you (and anyone else on the list) a copy of the reoprt with the comparison of the 63x vs the 100x lens if you want.
Cheers
Cam
Cameron Nowell B.Sc (Hons)
Microscopy Imaging and Research Core Facility Peter MacCallum Cancer Centre
7 St Andrews Place
East Melbourne, Victoria 3002
Phone: +61396561243
Mobile: +61422882700
Fax: +61396561411
________________________________
From: Confocal Microscopy List on behalf of Yang, Ann-Fook
Sent: Sat 27/10/2007 6:09 AM
To:
[hidden email]
Subject: Objective for confocal
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalHi All,
I am new in confocal microscopy.
We have a 40x/1.3 oil and a 63X/1.2 water objectives. Is there any advantage to add a 100X/1.46 Oil objective?
Ann Fook Yang,
Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada
EM Unit/ Unite EM
Edifice K.W. Neatby Building,
960 Carling Av /960 Boulevard Carling,
Ottawa,Ontario
K1A 0C6
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