Resolution. [from the New Merriam-Webster Dictionary, 1989 ed.]:

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Resolution [from the New Merriam-Webster Dictionary, 1989 ed.]: resolve v : 1 to break up into constituent parts: ANALYZE; 2 to find an answer to : SOLVE; 3 DETERMINE, DECIDE; 4 to make or pass a formal resolution resolution n : 1 the act or process of resolving 2 the action of solving, also : SOLUTION; 3 the quality of being resolute: FIRMNESS, DETERMINATION; 4 a formal statement expressing the opinion, will or, intent of a body of persons 05/04/09 wk13-a- 11

Rayleigh resolution limit 1 Total intensity Source 1 Source 2 1 Total intensity Source 1 Source 2 0.75 0.75 I [a.u.] 0.5 I [a.u.] 0.5 0.25 0.25 0 10 8 6 4 2 0 2 4 6 8 10 x [µm] 0 10 8 6 4 2 0 2 4 6 8 10 x [µm] Two point sources are well resolved if they are spaced such that: (i) the PSF diameter (i) the PSF radius equals the point source spacing equals the point source spacing 05/04/09 wk13-a-12

Diffraction limited resolution Two point objects are just resolvable (limited by diffraction only) if they are separated by: Two dimensional systems (rotationally symmetric PSF) One dimensional systems (e.g. slit like aperture) Safe definition: (one lobe spacing) Pushy definition: (1/2 lobe spacing) 05/04/09 wk13-a-13 You will see different authors giving different definitions. Rayleigh in his original paper (1879) noted the issue of noise and warned that the definition of just resolvable points is system or application dependent

Aberrations further limit resolution All our calculations have assumed geometrically perfect systems, i.e. we calculated the wave optics behavior of systems which, in the paraxial geometrical optics approximation would have imaged a point object onto a perfect point image. The effect of aberrations (calculated with non paraxial geometrical optics) is to blur the geometrically perfect image; including the effects of diffraction causes additional blur. 05/04/09 wk13-a-14 Fig. 9I in Jenkins, Francis A., and Harvey E. White. Fundamentals of Optics. 4th ed. New York, NY: McGraw-Hill, 1976. ISBN: 9780070323308. (c) McGraw-Hill. All rights reserved. This content is excluded from our Creative Commons license. For more information, see http://ocw.mit.edu/fairuse.

Aberration-limited resolution based on the MTF 1 wave optics picture 1 2u x,max diffraction limited (aberration free) 1D MTF 2u x,max 2u x,max 1D MTF with aberrations 2u x,max Fourier transform diffraction limited 1D PSF (sinc 2 ) Fourier transform something wider 05/04/09 wk13-a-15

Resolution: common misinterpretations Attempting to resolve object features smaller than the resolution limit (e.g. 1.22λ/NA) is hopeless. NO: resolution Image quality degradation as object features become smaller than the limit ( exceed the resolution limit ) is noise dependent and gradual. Besides, digital processing of the acquired images (e.g. methods such as the CLEAN algorithm, Wiener filtering, expectation maximization, etc.) can be employed. 05/04/09 wk13-a-16

Resolution: common misinterpretations Super-resolution By engineering the pupil function ( apodizing ) to result in a PSF with narrower side lobe, one can beat the resolution limitations imposed by the angular acceptance (NA) of the system. MAYBE: (i) (ii) Pupil engineering always results in narrower main lobe but accentuated side lobes lower power transmitted through the system Both effects can be BAD on the image 05/04/09 wk13-a-17

Pupil engineering example: apodization f 1 =20cm λ=0.5µm 05/04/09 wk13-a-18

Effect of apodization on the MTF and PSF Un-apodized Apodized: annular 05/04/09 wk13-a-19

Effect of apodization on the MTF and PSF Un-apodized Apodized: Gaussian 05/04/09 wk13-a-20

Pupil engineering trade-offs main lobe size sidelobes and vice versa main lobe size sidelobes generally, power loss means SNR degradation Annular type pupil functions typically narrow the main lobe of the PSF at the expense of higher side lobes Gaussian type pupil functions typically suppress the side lobes but broaden the main lobe of the PSF Compromise? application dependent for point like objects (e.g., stars) annular apodizers may be a good idea for low frequency objects (e.g., diffuse tissue) Gaussian apodizers may image with fewer artifacts Caveat: Gaussian amplitude apodizers very difficult to fabricate and introduce energy loss binary phase apodizers (lossless by nature) are used instead; typically designed by numerical optimization 05/04/09 wk13-a-21

Resolution: common misinterpretations This super cool digital camera has resolution of 8 Mega pixels (8 million pixels). NO: space bandwidth This is the most common and worst misuse of the term resolution. They are actually referring to the product (SBP) of the camera 05/04/09 wk13-a-22

Are resolution and number of pixels related? Answer depends on the magnification and PSF of the optical system attached to the camera PSF of optical system pixels on camera die Pixels significantly smaller than the system PSF are somewhat underutilized (the effective SBP is reduced) 05/04/09 wk13-a-23

Some more misstatements It is pointless to attempt to resolve beyond the Rayleigh criterion (however defined) NO: difficulty increases gradually as feature size shrinks, and difficulty is noise dependent Apodization can be used to beat the resolution limit imposed by the numerical aperture NO: watch sidelobe growth and power efficiency loss The resolution of my camera is N M pixels NO: the maximum possible SBP of your system may be N M pixels but you can easily underutilize it (i.e., achieve SBP that is less than N M) by using a suboptimal optical system 05/04/09 wk13-a-24

So, what is resolution? Our ability to resolve two point objects (in general, two distinct features in a more general object) based on the image however, this may be difficult to quantify Resolution is related to the NA but not exclusively limited by it Resolution, as it relates to NA: it s true that resolution improves as NA increases Other factors affecting resolution: caveats to the previous statement are aberrations / apodization (i.e., the exact shape of the PSF) NOISE! Is there an easy answer? No but when in doubt quote 0.61λ/(NA) or 1.22λ/(NA) as an estimate (not as an exact limit). 05/04/09 wk13-a-25

Today Two more applications of the Transfer Function defocus and Depth of Focus / Depth of Field (DoF) image reconstruction: deconvolution and its problems Tikhonov-regularized inverse filters Wednesday Polarization The intensity distribution near the focus of high-na imaging systems Utilizing the short depth of field of high-na imaging: confocal microscopy and related 3D imaging systems 05/11/09 wk14-a- 1

Defocus in wide field imaging Image removed due to copyright restrictions. Please see: http://www.imdb.com/media/rm4216035584/tt0137523 05/11/09 wk14-a- 2

Paraxial intensity distribution near focus x (rotationally symmetric wrt z axis) z Δx: Rayleigh resolution criterion [Lecture 23] Δz: Depth of Focus / Depth of Field (DoF) [today s topic] Note: at very high numerical apertures, the scalar approximation is no longer good; the vectorial nature of the electromagnetic field becomes important. 05/11/09 wk14-a- 3

4F system with in-focus input on-axis wave illumination NA a x max object transparency in-focus object pupil (Fourier) image Numerical Aperture 05/11/09 wk14-a- 4 in the paraxial approximation neglecting the low-pass filtering due to the finite pupil mask

4F system with out-of-focus input on-axis wave illumination δ object transparency out-of-focus object object convolved with the propagation kernel pupil (Fourier) object spectrum multiplied by the Fourier transform of the propagation kernel image 05/11/09 wk14-a- 5

Equivalent optical system on-axis wave illumination object transparency in-focus object phase mask represents defocus pupil (Fourier) image defocus ATF object spectrum multiplied by the complex transmissivity of an equivalent phase mask 05/11/09 wk14-a- 6

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