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In coherent imaging, complex wave amplitudes add with their relative wave phases preserved. The measured light intensity includes wave interference between the contributions.

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.