3/21/2023 0 Comments Diffraction limit![]() ![]() Imager Diagonal is larger than the Field Number of the Objective. Actual values mayĥ) Vignetting, a darkening of the image at the corners, may be observed if the (WD) and Field Number (FN) are shown for reference only. Maximum pixel size = R*magnification/2ģ) Camera and Objective combinations which meet the Nyquist criteria and are therefore diffraction limited are indicated by showing the Limiting Resolution in green font. R = 1.22*λ/(2NA)Įstimated by applying the Nyquist criteria to the minimum feature size as itĪppears on the imager plane. Minimum feature size is estimated using the Rayleigh Criteria at λ = 550nm it can be re-calculated for other wavelengths. Please contact our imaging experts for assistance in selecting a camera & objective combination that fulfils your FOV & resolution requirements at application-specific wavelengths. This table also applies to cooled sCMOS cameras such as the pco.edge 4.2, pco.edge 4.2bi and pco.edge 4.2bi uv cameras.Įstimated Performance of Camera and Objective Combinations In the table below, we show the Limiting Resolution (in µm) and the Field of View for several commercially available microscope objectives when used with a 2048 x 2048 camera with 6.5µm pixels: this includes non-cooled sCMOS cameras such as the pco.panda 4.2, pco.panda 4.2bi and the pco.panda 4.2bi uv. The above method can be used to estimate the performance of a camera and a microscope system, as long as the relevant parameters of the camera and the microscope objective are known. Using the Numerical Aperture of a microscope objective to estimate the Nyquist-limited maximum pixel size In the following table, “R” represents the Rayleigh Criterion for the diffraction-limited spot size: R = 1.22λ/(2NA) Objective This can be quantified by applying the Nyquist Criterion to the spatial sampling on the image plane, leading to an upper limit on the pixel size of the image sensor that is used in conjunction with a microscope. Lichtman then shows a sequence of images taken with cameras that have different pixel pitches providing a visual insight into why the sampling in the image plane must be sufficient to resolve an image. Our main takeaway is that the higher the NA, the smaller the Rayleigh Criterion and the better the resolution. We learn about the Rayleigh Criterion, and that it is different for the XY and XZ planes of a microscope. ![]() How close could two points be in the sample plane and still be resolved as separate points in the image. Lichtman leverages the previously explained concepts of diffraction, NA and PSF and builds up to the final payoff: optical resolution. ![]()
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