High sensitivity cameras can lower spatial resolution in high-resolution optical microscopy

Henning Ortkrass (), Marcel Müller, Anders Kokkvoll Engdahl, Gerhard Holst and Thomas Huser ()
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Henning Ortkrass: Bielefeld University
Marcel Müller: Bielefeld University
Anders Kokkvoll Engdahl: Bielefeld University
Gerhard Holst: Donaupark 11
Thomas Huser: Bielefeld University

Nature Communications, 2024, vol. 15, issue 1, 1-7

Abstract: Abstract High-resolution optical fluorescence microscopies and, in particular, super-resolution fluorescence microscopy, are rapidly adopting highly sensitive cameras as their preferred photodetectors. Camera-based parallel detection facilitates high-speed live cell imaging with the highest spatial resolution. Here, we show that the drive to use ever more sensitive, photon-counting image sensors in cameras can, however, have detrimental effects on the spatial resolution of the resulting images. This is particularly noticeable in applications that demand a high space-bandwidth product, where the image magnification is close to the Nyquist sampling limit of the sensor. Most scientists will often select image sensors based on parameters such as pixel size, quantum efficiency, signal-to-noise performance, dynamic range, and frame rate of the sensor. A parameter that is, however, typically overlooked is the sensor’s modulation transfer function (MTF). We have determined the wavelength-specific MTF of front- and back-illuminated image sensors and evaluated how it affects the spatial resolution that can be achieved in high-resolution fluorescence microscopy modalities. We find significant differences in image sensor performance that cause the resulting spatial resolution to vary by up to 28%. This result shows that the choice of image sensor has a significant impact on the imaging performance of all camera-based optical microscopy modalities.

Date: 2024
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DOI: 10.1038/s41467-024-53198-1

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