Let . After offset subtraction, each summed image has mean analogue-to-digital units per detector pixel. For a constant detector conversion gain and independent Poisson distribution detected counts, : summing independent frames adds both their means and variances. Here is the mean of the sum, not one frame.
Write , . At high counts, the delta method gives . Independence therefore givesThus the formula is a high-signal gain estimation from flat-field ratios, not an exact identity for a ratio of random variables. A Poisson distribution denominator can even be zero at low counts. The exposures must be linear and unsaturated, offsets removed, and read noise negligible. Being close to saturation increases the signal but does not establish linearity; clipping or charge-induced correlations biases a variance estimate. A large ensemble of suitable detector pixels supplies the measured standard deviation. In normal photodetector terminology is collected electrons per ADU, consistent with detected photons here only under the stated unit-yield convention; see the instrument gain definition.
A stable multiplicative sensitivity factor at detector pixel changes both means to . The noise-free ratio is still one, so flat-field correction is not required merely to remove that fixed pattern from the mean ratio. This is the principal advantage of using two matched flats rather than the variance of a single image.
However, the photon shot noise is not cancelled. At high counts the conditional ratio variance is . Pooling detector pixels gives , where brackets denote a spatial average. If the formula uses , it returnsThe inequality follows from the Jensen inequality for on positive signals. For small fractional response variation, the relative bias is approximately minus its squared coefficient of variation. A locally nearly uniform region or a fit using detector pixel-dependent variances avoids this bias from mixing the harmonic mean with the arithmetic mean. Stable response cancellation also presumes the conversion gain itself is uniform; gain variations, offset errors or changing sensitivity do not obey that simple cancellation. The fixed response pattern cancels in the ratio mean; unequal shot-noise levels can still bias a globally pooled gain estimate.
For independent reads in each sum, read noise contributes in squared ADU units. HenceUsing the shot-noise-only formula therefore givesThe condition for negligible contamination is , equivalently a large number of collected electrons per individual frame compared with . Summing more identical-level frames does not change that ratio because both and the read variance scale with . Measure read noise from paired bias images, or fit mean versus noise variance across several illumination levels: the shot-noise slope determines detector gain, while the read term gives an intercept. If is known, the corrected positive solution satisfies . Offset subtraction removes the mean bias but not the read variance. Correlated read errors need a different covariance calculation.
If spatial illumination is stable between the two sums, it acts just like the sensitivity factor: the mean ratio remains one but each detector pixel has a different shot-noise variance. A whole-image standard deviation is disproportionately influenced by faint detector pixels, so substituting a global arithmetic mean signal generally underestimates detector conversion gain. If the illumination pattern differs between the sums, their noise-free ratio is not constant; its structure adds apparent variance and causes a further systematic error.
Use locally uniform signal bins or subregions, remove offsets, mask defective or saturated detector pixels, and propagate the local shot and read variances. In the stable-pattern case a useful alternative to pooling raw ratios is the normalized differenceAt high signal and negligible read noise, treating the denominator at its mean gives , independent of the illumination. Thus a variance fit or can use a spatially varying flat without the arithmetic/harmonic mismatch. Measured noisy denominators cause higher-order corrections, so independent or smoothed signal estimates can be preferable. Fit and subtract a genuine changing illumination pattern before noise analysis, using sufficient smoothing not to fit away the random noise. Arbitrary flat-field normalization without propagating its noise does not by itself recover the original gain.
If temporal variation is spatially uniform, summing all frames in each group still gives independent Poisson distribution totals, but their means may now be with ratio . At high counts and negligible read noise, direct propagation givesAfter dividing the ratio image by its measured mean , the appropriate estimator isThe original formula is recovered only for . Uniform brightness drift does not automatically create a spatial pattern or extra ratio scatter beyond these changed noise levels; its leading problem is unequal group normalization. Interleave the groups in time or pair exposures of comparable flux, measure their separate means, and propagate shot and read noise when rescaling. If the spatial pattern or photodetector sensitivity also evolves, the ratio develops systematic structure that must be modeled or those frames rejected. A normalization inferred from finitely many detector pixels introduces a small shared correlation, which vanishes in the large-pixel limit and can be included in a precise uncertainty analysis.
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