Posted by
Stanislav Vitha-2 on
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When I did a test some years ago on our old Zeiss Axio system, using the phase contrast Plan Neofluar objective versus the non-phase objective of the same NA (Plan Neofluar or a Plan Apo, I do not remember) resulted in about 25 or 30% lower signal. This was true for a 40x/0.75 dry and a 100x/1.3 oil objectives.
I think one should expect that different manufacturers or even different types of phase contrast objectives (negative phase, positive phase, anoptral phase, ...) will have different combination of attenuation (and optionally a phase shift) on the phase ring, as well as phase shift in the area outside and inside the ring where the diffracted orders would pass through in Zernike phase contrast.
Stan Vitha
Microscopy and Imaging Center
Texas A&M University
On Mon, 4 May 2020 13:03:19 +0000, Knecht, David <
[hidden email]> wrote:
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>I suspect the higher number (20%) is if you consider loss in both excitation and emission through the phase objective, but in the excitation path, you usually have plenty of extra power to compensate for a bit of loss in excitation efficiency. So I think the lower number is more realistic.
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>Dr. David Knecht
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>University of Connecticut
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>On May 4, 2020, at 2:20 AM, Catalin Chiritescu <
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>*Message sent from a system outside of UConn.*
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>You lose about 20-30% of light when a Phase Contrast objective is used in fluorescence. AFAIK Nikon Ti (don't know about TI2) has an external phase ring setup that you can use for Phase Contrast imaging with BF objectives - you can take it out of fluorescence light path to avoid the loss and PSF disruption. We considered that option for our Quantitative Phase Imaging SLIM systems.