![Page 1: Objective Lens Markings and Interior Lenses...A digital image is acquired through a process o ften referred to as sampling. The accuracy of each sample depends upon the size of the](https://reader035.vdocuments.pub/reader035/viewer/2022062509/610320e0b20ffc1696787f7a/html5/thumbnails/1.jpg)
Objective Lens Markings and Interior Lenses
http://micro.magnet.fsu.edu/
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NA (Numerical Aperture) is EVERYTHING
http://micro.magnet.fsu.edu/
![Page 3: Objective Lens Markings and Interior Lenses...A digital image is acquired through a process o ften referred to as sampling. The accuracy of each sample depends upon the size of the](https://reader035.vdocuments.pub/reader035/viewer/2022062509/610320e0b20ffc1696787f7a/html5/thumbnails/3.jpg)
http://micro.magnet.fsu.edu/
NA and Resolution
Abbe = d = __λ___2 NA
Rayleigh = d = 0.61NA
λ
See also http://www.olympusfluoview.com/java/resolution3d/index.html
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Dec
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Increase NA
=Increase resolution
http://micro.magnet.fsu.edu/
Effects of Wavelength and NA on Resolution
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http://micro.magnet.fsu.edu/
Air vs. Oil Objectives
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http://micro.magnet.fsu.edu/
Effects of Confocal Pinhole Aperture Sizes
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A digital image is acquired through a process often referred to as sampling. The accuracy of each sample depends upon the size of the sampling interval, which is determined by the number of pixels in the image and the distance between each pixel. Each image sample is taken as an average of the image brightness over the sampling interval. The Nyquist Criterion requires a sampling interval equal to just over twice (actually 2.3 times) the highest spatial frequency occurring in the image to avoid losing spatial information. If the sampling interval is larger than the Nyquist limit, then undersampling occurs, and spatial information is lost. Undersampling has the effect of distorting image details, resulting in a phenomenon termed aliasing, which occurs when undersampled high spatial frequencies masquerade as (or "alias" to) lower spatial frequencies.
http://micro.magnet.fsu.edu/
Nyquist Sampling Criterion
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http://micro.magnet.fsu.edu/
Nyquist Criterion: Acquire samples at 2.3 X the highest spatial frequency (resolution)in the image
Nyquist Sampling Criterion
![Page 9: Objective Lens Markings and Interior Lenses...A digital image is acquired through a process o ften referred to as sampling. The accuracy of each sample depends upon the size of the](https://reader035.vdocuments.pub/reader035/viewer/2022062509/610320e0b20ffc1696787f7a/html5/thumbnails/9.jpg)
http://micro.magnet.fsu.edu/
Nyquist Sampling Criterion