3D test sample for the calibration and quality control of stimulated emission depletion (STED) and confocal microscopes

Ernest B. van der Wee; Jantina Fokkema; Chris Kennedy; Marc del Pozo Puig; D.A.M.  de Winter; Peter N.A. Speets; Hans C. Gerritsen; Alfons van Blaaderen

doi:10.1038/s42003-021-02432-3

3D test sample for the calibration and quality control of stimulated emission depletion (STED) and confocal microscopes

Ernest B. van der Wee
, Jantina Fokkema
, Chris Kennedy
, Marc del Pozo Puig
, D.A.M. de Winter
, Peter N.A. Speets
, Hans C. Gerritsen
, Alfons van Blaaderen (Corresponding author)

Research output: Contribution to journal › Article › Academic › peer-review

8 Citations (Scopus)

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Abstract

Multiple samples are required to monitor and optimize the quality and reliability of quantitative measurements of stimulated emission depletion (STED) and confocal microscopes. Here, we present a single sample to calibrate these microscopes, align their laser beams and measure their point spread function (PSF) in 3D. The sample is composed of a refractive index matched colloidal crystal of silica beads with fluorescent and gold cores. The microscopes can be calibrated in three dimensions using the periodicity of the crystal; the alignment of the laser beams can be checked using the reflection of the gold cores; and the PSF can be measured at multiple positions and depths using the fluorescent cores. It is demonstrated how this sample can be used to visualize and improve the quality of STED and confocal microscopy images. The sample is adjustable to meet the requirements of different NA objectives and microscopy techniques and additionally can be used to evaluate refractive index mismatches as a function of depth quantitatively

Original language	English
Article number	909
Number of pages	9
Journal	Communications biology
Volume	4
DOIs	https://doi.org/10.1038/s42003-021-02432-3
Publication status	Published - 23 Jul 2021
Externally published	Yes

Funding

Funders	Funder number
Netherlands Center for Multiscale Catalytic Energy Conversion
Seventh Framework Programme	291667, FP-2007-2013
European Union's Horizon 2020 - Research and Innovation Framework Programme
Ministerie van Onderwijs, Cultuur en Wetenschap
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	12715

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10.1038/s42003-021-02432-3Licence: CC BY

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title = "3D test sample for the calibration and quality control of stimulated emission depletion (STED) and confocal microscopes",

abstract = "Multiple samples are required to monitor and optimize the quality and reliability of quantitative measurements of stimulated emission depletion (STED) and confocal microscopes. Here, we present a single sample to calibrate these microscopes, align their laser beams and measure their point spread function (PSF) in 3D. The sample is composed of a refractive index matched colloidal crystal of silica beads with fluorescent and gold cores. The microscopes can be calibrated in three dimensions using the periodicity of the crystal; the alignment of the laser beams can be checked using the reflection of the gold cores; and the PSF can be measured at multiple positions and depths using the fluorescent cores. It is demonstrated how this sample can be used to visualize and improve the quality of STED and confocal microscopy images. The sample is adjustable to meet the requirements of different NA objectives and microscopy techniques and additionally can be used to evaluate refractive index mismatches as a function of depth quantitatively",

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doi = "10.1038/s42003-021-02432-3",

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AU - van der Wee, Ernest B.

AU - Fokkema, Jantina

AU - Kennedy, Chris

AU - del Pozo Puig, Marc

AU - de Winter, D.A.M.

AU - Speets, Peter N.A.

AU - Gerritsen, Hans C.

AU - van Blaaderen, Alfons

PY - 2021/7/23

Y1 - 2021/7/23

N2 - Multiple samples are required to monitor and optimize the quality and reliability of quantitative measurements of stimulated emission depletion (STED) and confocal microscopes. Here, we present a single sample to calibrate these microscopes, align their laser beams and measure their point spread function (PSF) in 3D. The sample is composed of a refractive index matched colloidal crystal of silica beads with fluorescent and gold cores. The microscopes can be calibrated in three dimensions using the periodicity of the crystal; the alignment of the laser beams can be checked using the reflection of the gold cores; and the PSF can be measured at multiple positions and depths using the fluorescent cores. It is demonstrated how this sample can be used to visualize and improve the quality of STED and confocal microscopy images. The sample is adjustable to meet the requirements of different NA objectives and microscopy techniques and additionally can be used to evaluate refractive index mismatches as a function of depth quantitatively

AB - Multiple samples are required to monitor and optimize the quality and reliability of quantitative measurements of stimulated emission depletion (STED) and confocal microscopes. Here, we present a single sample to calibrate these microscopes, align their laser beams and measure their point spread function (PSF) in 3D. The sample is composed of a refractive index matched colloidal crystal of silica beads with fluorescent and gold cores. The microscopes can be calibrated in three dimensions using the periodicity of the crystal; the alignment of the laser beams can be checked using the reflection of the gold cores; and the PSF can be measured at multiple positions and depths using the fluorescent cores. It is demonstrated how this sample can be used to visualize and improve the quality of STED and confocal microscopy images. The sample is adjustable to meet the requirements of different NA objectives and microscopy techniques and additionally can be used to evaluate refractive index mismatches as a function of depth quantitatively

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DO - 10.1038/s42003-021-02432-3

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JO - Communications biology

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ER -

3D test sample for the calibration and quality control of stimulated emission depletion (STED) and confocal microscopes

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Erratum: Publisher Correction: 3D test sample for the calibration and quality control of stimulated emission depletion (STED) and confocal microscopes (Communications biology (2021) 4 1 (909))

Cite this