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Showing content from https://pubmed.ncbi.nlm.nih.gov/12801419/ below:

ScanImage: flexible software for operating laser scanning microscopes

Image Quality in ScanImage A)

Apparent quantum efficiency as a function of sampling rate in

ScanImage

for a single-photon-pulse with a 2.35 μs FWHM and unit amplitude. The red trace shows the apparent quantum efficiency (

) of the signal (see inset) determined by digital integration. The black trace shows

sampled at 200 kHz to 5 MHz.

approaches 1 as the sample rate approaches 1 MHz. The blue points are the normalized apparent quantum efficiencies from an actual PMT illuminated by a constant light source (data pre-filtered at 330 kHz).

ScanImage

uses a sample rate of 1.25 MHz (arrow), which yields nearly ideal signal-to-noise

≈ 1. This analysis is only valid for pulses that are al least 2.35 μs in duration.

Inset:

20-μs pulse interval comprised of 2.35-μs events sampled at different rates.

B)

Histogram of the events in the pixel interval (bars) and Poisson fit (line).

C)

Model of bleedthrough of photons from adjacent pixels. A single-photon-pulse with a 2.5 μs FWHM was placed randomly in pixel intervals of widths 2 to 10 μs. The number of trials at each pixel interval was 10,000. The percent bleedthrough equaled 100*(1 - IPP/IEP) where IEP is the integral of the entire pulse and IPP is the integral of the region of the pulse that fell in the pixel interval. Shown is the mean value of the percent bleedthrough for each pixel time normalized with respect to the pulse width (Pixel Interval/Pulse Width). The error bars (SEM) were smaller than the marker points and were omitted.


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