Quality Control of Fluorescence Imaging Systems

Abstract

We have developed a new tool for the assessment and monitoring of most of the performances of fluorescence microscopes. We believe it can advantageously be integrated in the quality control process of core facilities where a certain level of performance for the end users must be assured.

Fluorescence calibration standards made from broadband emitters encapsulated in polymer beads for fluorescence microscopy and flow cytometry

“…sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/ Datasheet/10/75194dat.pdf). Also for fluorescence microscopy, in addition to a very small number of relatively expensive structured calibration tools like the slide from Argolight [21,22], different calibration beads have been commercialized to determine parameters like resolution x/y/ z, intensity calibration, color adjustment, instrument alignment, and stability [20,23,24] (https://www. thermofisher.com/de/de/home/references/newsletters-andjournals/bioprobes-journal-of-cell-biology-applications/ bioprobes-70/fluorescent-microspheres-for-calibration.html).…”

“…These calibration tools are intended to facilitate the assessment of instrument performance to ensure reliable measurements and to improve the comparability of FCM experiments [ 16 – 20 , 22 ] ( https://www.thermofisher.com/de/de/home/references/newsletters-and-journals/bioprobes-journal-of-cell-biology-applications/bioprobes-70/fluorescent-microspheres-for-calibration.html ), ( https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Datasheet/10/75194dat.pdf ). Also for fluorescence microscopy, in addition to a very small number of relatively expensive structured calibration tools like the slide from Argolight [ 21 , 22 ], different calibration beads have been commercialized to determine parameters like resolution x / y / z , intensity calibration, color adjustment, instrument alignment, and stability [ 20 , 23 , 24 ] ( https://www.thermofisher.com/de/de/home/references/newsletters-and-journals/bioprobes-journal-of-cell-biology-applications/bioprobes-70/fluorescent-microspheres-for-calibration.html ). However, these broadly used beads have been neither designed nor yet used to determine the spectral characteristics of fluorescence measuring devices, like spectral scanning fluorescence microscopes, spectral FCM setups, microtiter plate readers, and all types of integral measuring devices, although for fluorescence microscopic techniques and even FCM, new trends in instrument design focus increasingly on spectrally resolved measurements [ 25 , 26 ].…”

A comparison of high resolution image reconstruction methods from confocal array detector with a new deconvolution algorithm

“…To estimate spatial resolution, we applied the eight reconstruction methods to the Argolight calibration slide [16] depicting vertical lines pattern (see Fig. 4a).…”

“…4b). As denoted in the calibration slide documentation [16] a measured contrast below 26.5% means that the objects at the line resolution cannot be distinguished. In our experiments, the PC, ISM and IFED methods cannot separate two lines less than 350 nm apart.…”

“…In our experiments, we compared the performance of PC, ISM, IFED, IS-FED, ISM-RL, ISM-W, ISM-SV and SV algorithms on several stacks of N T = 32 sub-detectors. First, we applied the eight reconstructions methods to a stack depicting a ring shape with homogeneous intensity, acquired with the Argolight calibration slide [16]. In Fig.…”