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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.

In incoherent imaging, contributions from distinct object points add as light intensities because their mutual spatial optical coherence is negligible. A single convolution additionally requires translation invariance of the point spread function.

The optical transfer function is the Fourier transform of the normalized intensity point spread function for incoherent imaging with translation invariance, using spatial frequency in cycles per unit length. It has $H(0)=1$. For an ideal circular pupil it is the normalized pupil autocorrelation, with cutoff $2\mathrm{NA}/{\lambda }_0$ in object coordinates: no overlap remains when two pupil copies are shifted apart by more than one pupil diameter.^[ref]

The Rayleigh criterion gives a separation in the sensor plane $\delta =f\Delta \theta =2.44\mu \mathrm{m}$, using the $5\mathrm{mm}$ image distance. Counting one square pixel per such spatial resolution element givesThis is an estimate of the number of optically distinguishable elements. To sample all the spatial information, specify a signal sampling criterion as well. For incoherent imaging through an ideal circular aperture, the optical transfer function has cutoff spatial frequency ${\nu }_c=D/(\lambda f)$. The Nyquist–Shannon sampling theorem requires a pixel pitchA common intermediate rule, two pixels per Rayleigh separation, gives about $8.1$ million pixels; it is a heuristic and does not quite sample the full ideal optical transfer function bandwidth. Thus about $2$ million is the elementary spatial resolution element count, while $12$ million is the ideal monochrome Nyquist–Shannon sampling theorem answer to “full use.” Color-filter layout and finite pixel area would impose further practical conditions.

A finite optical pupil spreads a point into a point spread function. In incoherent imaging an extended intensity distribution is convolved with that function; its Fourier transform is the optical transfer function, which attenuates and ultimately removes high spatial frequencies. For an ideal circular pupil the point response is an Airy disk. The Rayleigh criterion for two equally bright incoherent points isA microscope's object-space scale is of order $\lambda /(2\mathrm{NA})$, with the numerical factor depending on illumination and the criterion; $\mathrm{NA}=n\sin \alpha$ measures the accepted ray cone. A larger pupil or shorter wavelength improves the diffraction scale. Aberrations, defocus, detector pixel sampling, motion, background and photon noise can worsen practical performance. Merely magnifying the same blurred image adds no information. Coherent sources require amplitude superposition instead of the incoherent intensity rule. Rayleigh resolution is a convention for separating a pair, distinct from the precision of localizing one isolated point; information does not abruptly vanish below it.

A point spread function is an imaging system's response to an optical point source. Here it means the distribution of image light intensity: an ideal point object becomes a finite spot, often with faint rings, because of diffraction. An unresolved star and a sufficiently small bead emitting fluorescence are practical approximations to an optical point source. Their image tells us how nearby object details will overlap, and reveals blurring from optical aberrations, defocus, and, for an optical telescope, astronomical seeing.

A useful convention separates shape from throughput. At a fixed image plane, let $p(\mathbf{x})\ge 0$ be the point spread function with ${\int }_{{\mathbb{R}}^2}p(\mathbf{x})d^2\mathbf{x}=1$. Its units are inverse area. A point delivering detected radiant flux $F$ at image position ${\mathbf{x}}_0$ produces light intensity $Fp(\mathbf{x}-{\mathbf{x}}_0)$; this point spread function normalization makes the integral over the image equal to $F$. A profile divided by its own peak is convenient for comparing widths, but is a different point spread function normalization and does not by itself compare total radiant flux.

In incoherent imaging, different object points have negligible mutual spatial optical coherence over the measurement, so their image light intensities add. If the response also obeys translation invariance over the chosen field of view, the expected continuous image is the convolutionHere $O$ is the object brightness distribution expressed in image-scaled coordinates, with magnification already included, and $\tau$ is constant transmission. For $O(\mathbf{u})=F_0\delta (\mathbf{u}-{\mathbf{u}}_0)$, where $\delta$ is the two-dimensional Dirac delta function, this gives $I=\tau F_{0}p(\mathbf{x}-{\mathbf{u}}_0)$. A changing response across the field of view instead requires a position-dependent integral kernel $p(\mathbf{x};\mathbf{u})$; one measured point spread function is then only a local calibration.

In coherent imaging, the superposition principle applies to the complex wave amplitudes. With amplitude spread function $h$, the corresponding ideal model isThus $|h\ast E{|}^2$ generally differs from $|h{|}^2\ast |E{|}^2$: coherent contributions produce wave interference terms that depend on their relative wave phases. Their time average vanishes for mutually incoherent emitters, which explains the intensity convolution. Partial optical coherence requires the correlations between object points as well.^[ref]

The figure is an original diffraction model, not measured data. All panels have the same total radiant flux: each member of a pair contributes half the single-source radiant flux. Dashed curves show the separate light intensity contributions; the solid curve is their sum. The point spread functions have tails, so even sources satisfying the Rayleigh criterion overlap. As separation decreases, the image changes continuously; there is no abrupt disappearance of all information at one separation.

A motivating measured example is Measurement of point-spread function (PSF) for confocal fluorescence microscopy by InCheon Song and others (2005). A $543.8\mathrm{nm}$ laser, delivered through an optical fiber, excited fluorescent beads on a slide through a $100\times$, numerical aperture $1.3$ oil-immersion objective. A moving stage scanned the bead; emitted light returned through the objective, an emission filter and a pinhole to a photodiode. The readout was fluorescence light intensity against scanned position. Under the same setup, nominal bead diameters $0.17$, $0.5$, and $1\mu \mathrm{m}$ produced progressively broader measured spots. The size comparison controls for the finite object: the measured response is the convolution of bead emission with the effective point spread function, rather than a pure point response. Inverting this convolution model gave a response compared with the theoretical one. The smaller bead gave a weaker signal relative to noise; uncertainty in actual bead diameter also affected the restored width. These are limits of the calibration. In confocal microscopy, excitation and pinhole detection jointly determine the effective point spread function; this experiment does not measure the ideal widefield Airy disk plotted above.^[ref]

For a new calibration, useful additional controls are to measure detector background, avoid saturating the photodetector, repeat the scan to check stability, and compare isolated beads at several positions in the field of view. Record wavelength, numerical aperture, pinhole setting, sampling interval and optical focus. These are practical recommendations, not extra measurements claimed for the published experiment.

The Rayleigh criterion calls two equal, mutually incoherent optical point sources just resolved when one circular-pupil Airy disk's centre coincides with the other's first zero. It is a specified discrimination convention, distinct from full width at half maximum and from precision in estimating one source's position.

For this circular optical pupil, the Rayleigh criterion uses angular separation $s_{\mathrm{R}}={\theta }_1\simeq 1.22\lambda /D$, or $s_{\mathrm{R}}=r_1\simeq 1.22\lambda f/D$ in the paraxial approximation to the focal plane. The point spread functions still overlap.^[ref]

For the Airy disk design example with $D=5.00\mathrm{mm}$, $f=100\mathrm{mm}$ and $\lambda =550\mathrm{nm}$, the angular convention gives $s_{\mathrm{R}}=1.34164\times 10^{-4}\mathrm{rad}$, or about $27.67$ arcseconds, and the image-plane separation is $13.4164\mu \mathrm{m}$. Two equally bright, mutually incoherent optical point sources at separations $0.7$, $1$ and $1.5$ times this scale would have image separations $9.391$, $13.416$ and $20.125\mu \mathrm{m}$ respectively. These calculated cases are compared in incoherent two-source Airy profile.

The Rayleigh criterion is useful because it ties an easily recognized two-peak convention to a predictable optical scale. It does not assert that all information vanishes below that separation. Inferring two unknown positions also depends on photon shot noise, background, unequal brightness and uncertainty in the point spread function; locating a single isolated source is a different task. A theoretical analysis shows that alternative optical measurements can estimate the separation of two incoherent optical point sources below this scale.^[ref]

The mutual optical coherence condition is essential. In incoherent imaging, time-averaged light intensities add. In coherent imaging, wave amplitudes add before squaring, leaving a wave interference term controlled by relative wave phase. Two holes illuminated by the same coherent laser therefore do not implement the equal-incoherent-source convention merely because their individual patterns resemble Airy disks. The coherent single-aperture laboratory test establishes the response of each aperture; it does not establish the pair's mutual incoherence.