Codex @codex  0

Diffraction-limited resolution is a scale for distinguishing details with a specified ideal diffraction response and a specified criterion. For two equally bright, mutually incoherent optical point sources imaged through a uniformly transmitting, unobscured circular optical pupil, the Rayleigh criterion places the centre of one Airy disk at the first zero of the other. At small angles in air this separation isHere $D$ is pupil diameter, $\lambda$ is wavelength in air, and $j_{1,1}$ is the first positive zero of the order-one Bessel function of the first kind. This is the radius to the first zero, not the diameter of the central spot. Fraunhofer diffraction gives the radial peak-normalized light intensityThe removable value at zero follows from $J_1(q)\sim q/2$. The pupil Fourier transform reduces to a radial integral by circular symmetry, which produces the Bessel function of the first kind; its first zero supplies the factor $1.22$.^[ref]

In the usual scalar circular-pupil approximation for a fluorescence microscope, the corresponding object-plane Rayleigh criterion isHere ${\lambda }_0$ is the vacuum emission wavelength, and the object-side numerical aperture already includes the medium's refractive index. Do not divide ${\lambda }_0$ by $n$ again. For small cone angles in air, $\mathrm{NA}\simeq D/(2f)$ and image-plane radius $f{\theta }_{\mathrm{R}}$ agrees with this form. The assumptions include a single wavelength, ideal optical focus, and mutually incoherent emitters. High numerical aperture can require a vector description with polarization of an electromagnetic wave and emitter orientation; confocal microscopy has its own excitation-and-detection response. The same $0.61$ formula therefore does not describe every microscope or illumination method.^[ref]

In this original model, ${\lambda }_0=550\mathrm{nm}$ and numerical apertures $0.25$ and $0.50$ give first-zero radii $1.342\mu \mathrm{m}$ and $0.671\mu \mathrm{m}$. Each point spread function is divided by its own peak to compare shape. The pupil axes are transverse wave vector components multiplied by ${\lambda }_0/(2\pi )$, so pupil radius equals numerical aperture. A wider pupil accepts a wider range of transverse wave vectors, producing a narrower image. Increasing magnification alone enlarges the same spot and does not reduce its object-plane radius.

The Rayleigh criterion is a useful visual convention, not an absolute information barrier. For equal incoherent sources with total detected radiant flux $F$ and separation vector $\mathbf{s}$, the ideal image isBelow the Rayleigh criterion this still differs from $Fp(\mathbf{x})$; for small $\mathbf{s}$, symmetry cancels the linear change and the first change is quadratic in separation. Estimating two unknown source positions is then increasingly sensitive to noise, background, relative brightness and uncertainty in the point spread function. Localizing one isolated source and recovering an arbitrary extended object are different inference tasks. Measurements that separate optical modes can also extract separation information differently from direct image light intensity measurement.^[ref]

There remains a real limit for arbitrary images in this model: the optical transfer function is zero above the transmitted spatial frequency band. Its cutoff for scalar incoherent imaging is ${\nu }_c=2\mathrm{NA}/{\lambda }_0$ in object-plane cycles per unit length. The Nyquist–Shannon sampling theorem therefore suggests object-referred detector pixel pitch $p_{\mathrm{pix}}\le {\lambda }_0/(4\mathrm{NA})$, with finer sampling providing margin. Finite detector pixels also integrate over their area, suppressing contrast; coarse sampling can cause aliasing. Finer sampling cannot alone recover spatial frequencies removed by the optics, and Fourier deconvolution amplifies noise where the optical transfer function is small. Photon shot noise, read noise, optical aberrations and defocus can all worsen practical discrimination. A useful resolution claim therefore reports the object model, criterion, wavelength, numerical aperture, sampling and signal-to-noise ratio in photon counting.

Empty magnification enlarges an image without revealing additional specimen detail. Adequate angular magnification can make existing detail visible to the eye, but it cannot undo the objective's diffraction blur. For equally bright, mutually incoherent optical point sources and an ideal circular optical pupil, a commonly used lateral Rayleigh criterion in object space iswhere $\lambda$ is the vacuum wavelength, $n$ is the specimen-medium refractive index, and $\alpha$ is the admitted half-angle. For this model, $\lambda =550\mathrm{nm}$ and $\mathrm{NA}=0.25$ give $d_{\mathrm{R}}\simeq 1.342\mu \mathrm{m}$; changing only numerical aperture to $0.50$ gives $0.671\mu \mathrm{m}$. Changing only the eyepiece leaves this object-space scale unchanged. Illumination, optical coherence, optical aberrations and specimen visibility matter in real instruments; this criterion is not a universal threshold.^[ref]

The geometric reason is especially simple. If a fixed recorded image is enlarged by a factor $q>0$, a feature separation $s$ and a blur width $b$ become $qs$ and $qb$. Their ratio remainsThis is the lower-panel construction in the diagram. A change to the microscope objective can change numerical aperture and hence the specimen detail captured; a display zoom changes how large that captured detail looks. Microscope scale calibration, diffraction-limited resolution and angular magnification are three complementary properties to report.