Quality Assurance Testing for Modern Optical Imaging Systems

Stack, Robert F.; Bayles, Carol J.; Girard, Anne-Marie; Martin, Karen H.; Opansky, C.; Schulz, Katherine; Cole, Richard W.

doi:10.1017/s1431927611000237

Cited by 29 publications

(36 citation statements)

References 11 publications

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“…Regular system checks should be common practice and measure e.g. the point spread function (PSF), the laser power, the illumination homogeneity of the field of view, the detector sensitivity as signal‐to‐noise‐ratio (SNR) and the coefficient of variation (Cole et al, 2011, 2013; Gelman and Rietdorf, 2010; Hibbs, 2006; Stack et al, 2011; Theer et al, 2014; Zucker and Price, 2001, 1999). When performed regularly, standardized test series often help identify damaged and misaligned microscope components, even before they significantly impair the quality of user data.…”

Section: How To Design Set Up and Run Your Imaging Core Facilitymentioning

confidence: 99%

Advanced light microscopy core facilities: Balancing service, science and career

Ferrando‐May

Hartmann

Reymann

et al. 2016

Microsc. Res. Tech.

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Core Facilities (CF) for advanced light microscopy (ALM) have become indispensable support units for research in the life sciences. Their organizational structure and technical characteristics are quite diverse, although the tasks they pursue and the services they offer are similar. Therefore, throughout Europe, scientists from ALM‐CFs are forming networks to promote interactions and discuss best practice models. Here, we present recommendations for ALM‐CF operations elaborated by the workgroups of the German network of ALM‐CFs, German Bio‐Imaging (GerBI). We address technical aspects of CF planning and instrument maintainance, give advice on the organization and management of an ALM‐CF, propose a scheme for the training of CF users, and provide an overview of current resources for image processing and analysis. Further, we elaborate on the new challenges and opportunities for professional development and careers created by CFs. While some information specifically refers to the German academic system, most of the content of this article is of general interest for CFs in the life sciences. Microsc. Res. Tech. 79:463–479, 2016. © 2016 THE AUTHORS MICROSCOPY RESEARCH AND TECHNIQUE PUBLISHED BY WILEY PERIODICALS, INC.

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Section: How To Design Set Up and Run Your Imaging Core Facilitymentioning

confidence: 99%

Advanced light microscopy core facilities: Balancing service, science and career

Ferrando‐May

Hartmann

Reymann

et al. 2016

Microsc. Res. Tech.

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“…Most of these problems would not have been noticed by the average facility users with their biological samples. And the confocal microscope cannot be examined in isolation either, the impact of environmental factors like air flow and temperature changes also needs to be considered, for example by measuring room and/or microscope stage temperature in conjunction with some of the performance tests (Figure S2A/B, Figure S10B/C)[29,37]. …”

Section: Resultsmentioning

confidence: 99%

ConfocalCheck - A Software Tool for the Automated Monitoring of Confocal Microscope Performance

Hng

Dormann

2013

PLoS ONE

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Laser scanning confocal microscopy has become an invaluable tool in biomedical research but regular quality testing is vital to maintain the system’s performance for diagnostic and research purposes.Although many methods have been devised over the years to characterise specific aspects of a confocal microscope like measuring the optical point spread function or the field illumination, only very few analysis tools are available. Our aim was to develop a comprehensive quality assurance framework ranging from image acquisition to automated analysis and documentation. We created standardised test data to assess the performance of the lasers, the objective lenses and other key components required for optimum confocal operation.The ConfocalCheck software presented here analyses the data fully automatically. It creates numerous visual outputs indicating potential issues requiring further investigation. By storing results in a web browser compatible file format the software greatly simplifies record keeping allowing the operator to quickly compare old and new data and to spot developing trends.We demonstrate that the systematic monitoring of confocal performance is essential in a core facility environment and how the quantitative measurements obtained can be used for the detailed characterisation of system components as well as for comparisons across multiple instruments.

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“…You should use a fluorescent plastic test slide in order to measure the light source intensity during warm up and the intensity stability over time to ensure it does not vary during your experimental timeframe (55). Some light sources may need more time to warm up (mercury arc lamps), while others may not need to be warmed up at all (LED light sources).…”

Section: Image Collection Corrections and Displaymentioning

confidence: 99%

Epi-Fluorescence Microscopy

Webb

Brown

2012

Methods in Molecular Biology

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Epi-fluorescence microscopy is available in most life sciences research laboratories, and when optimized can be a central laboratory tool. In this chapter, the epi-fluorescence light path is introduced and the various components are discussed in detail. Recommendations are made for incident lamp light sources, excitation and emission filters, dichroic mirrors, objective lenses, and charge-coupled device (CCD) cameras in order to obtain the most sensitive epi-fluorescence microscope. The even illumination of metal-halide lamps combined with new “hard” coated filters and mirrors, a high resolution monochrome CCD camera, and a high NA objective lens are all recommended for high resolution and high sensitivity fluorescence imaging. Recommendations are also made for multicolor imaging with the use of monochrome cameras, motorized filter turrets, individual filter cubes, and corresponding dyes that are the best choice for sensitive, high resolution multicolor imaging. Images should be collected using Nyquist sampling and should be corrected for background intensity contributions and nonuniform illumination across the field of view. Photostable fluorescent probes and proteins that absorb a lot of light (i.e., high extinction co-efficients) and generate a lot of fluorescence signal (i.e., high quantum yields) are optimal. A neuronal immune-fluorescence labeling protocol is also presented. Finally, in order to maximize the utility of sensitive wide-field microscopes and generate the highest resolution images with high signal-to-noise, advice for combining wide-field epi-fluorescence imaging with restorative image deconvolution is presented.

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Quality Assurance Testing for Modern Optical Imaging Systems

Cited by 29 publications

References 11 publications

Advanced light microscopy core facilities: Balancing service, science and career

Advanced light microscopy core facilities: Balancing service, science and career

ConfocalCheck - A Software Tool for the Automated Monitoring of Confocal Microscope Performance

Epi-Fluorescence Microscopy

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