Evaluation and Purchase of Confocal Microscopes: Numerous Factors to Consider

Zucker, Robert M.; Chua, Michael

doi:10.1002/0471142956.cy0216s54

Cited by 7 publications

(3 citation statements)

References 21 publications

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“…Currently, microscopy using advanced spectral detection methods are based on scanning through the emission spectra or static capture of the entire emission range with multiple detector arrays for each wavelength [27]. Both methods are capable of distinguishing the difference from FITC and EGFP, which have very similar emission spectra, separated by only about 20 nm.…”

Section: Microscopy Techniques and Visualization Of Single Cellsmentioning

confidence: 99%

“…The conventional method of using spectral filters has limitations in separating multiple, closely located emission spectra and, in the case of low-level fluorescence detection, is especially sensitive to instrumental artefacts such as dark-current (electronic noise) produced by the detector [ 26 ]. Currently, microscopy using advanced spectral detection methods are based on scanning through the emission spectra or static capture of the entire emission range with multiple detector arrays for each wavelength [ 27 ]. Both methods are capable of distinguishing the difference from FITC and EGFP, which have very similar emission spectra, separated by only about 20 nm.…”

Section: Microscopy Techniques and Visualization Of Single Cellsmentioning

confidence: 99%

See 1 more Smart Citation

Quantitative biology of single neurons

Eberwine

Lovatt

Buckley

et al. 2012

J. R. Soc. Interface.

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The building blocks of complex biological systems are single cells. Fundamental insights gained from single-cell analysis promise to provide the framework for understanding normal biological systems development as well as the limits on systems/cellular ability to respond to disease. The interplay of cells to create functional systems is not well understood. Until recently, the study of single cells has concentrated primarily on morphological and physiological characterization. With the application of new highly sensitive molecular and genomic technologies, the quantitative biochemistry of single cells is now accessible.

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Section: Microscopy Techniques and Visualization Of Single Cellsmentioning

confidence: 99%

Section: Microscopy Techniques and Visualization Of Single Cellsmentioning

confidence: 99%

Quantitative biology of single neurons

Eberwine

Lovatt

Buckley

et al. 2012

J. R. Soc. Interface.

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“…For the systematic analysis of confocal performance a wide range of tests have been developed, mostly relying on inexpensive samples and yet providing a wealth of information on the state of the instrument and individual components [7,8,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29]. Although there are many differences in the design of the commercially available confocal microscopes - and each of them available with a multitude of possible configurations - the following parameters are commonly checked for quality control purposes: …”

Section: Introductionmentioning

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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Evaluating Confocal Microscopy System Performance

Zucker

2013

Confocal Microscopy

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A confocal microscope was evaluated with a series of tests that measure field illumination, lens clarity, laser power, laser stability, dichroic functionality, spectral registration, axial resolution, scanning stability, PMT quality, overall machine stability, and system noise. These tests will help investigators measure various parameters on their confocal microscopes to insure that they are working correctly with the necessary resolution, sensitivity, and precision. Utilization of this proposed testing approach will help eliminate some of the subjectivity currently employed in assessing the CLSM performance.

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Evaluation and Purchase of Confocal Microscopes: Numerous Factors to Consider

Abstract: The purchase of a confocal microscope is a difficult decision. Many factors need to be considered, which include hardware, software, company, support, service, and price. These issues are discussed to help guide the purchasing process.

Cited by 7 publications

References 21 publications

Quantitative biology of single neurons

Quantitative biology of single neurons

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

Evaluating Confocal Microscopy System Performance

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