Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope

Pilger C, Pospisil J, Müller M, Ruoff M, Schuette M, Spiecker H, Huser T (2021)
Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences 379(2199): 20200300.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Autor*in
Pilger, ChristianUniBi; Pospisil, JakubUniBi; Müller, MarcelUniBi; Ruoff, Martin; Schuette, Martin; Spiecker, Heinrich; Huser, ThomasUniBi
Abstract / Bemerkung
Fluorescence-based microscopy as one of the standard tools in biomedical research benefits more and more from super-resolution methods, which offer enhanced spatial resolution allowing insights into new biological processes. A typical drawback of using these methods is the need for new, complex optical set-ups. This becomes even more significant when using two-photon fluorescence excitation, which offers deep tissue imaging and excellent z-sectioning. We show that the generation of striped-illumination patterns in two-photon laser scanning microscopy can readily be exploited for achieving optical super-resolution and contrast enhancement using open-source image reconstruction software. The special appeal of this approach is that even in the case of a commercial two-photon laser scanning microscope no optomechanical modifications are required to achieve this modality. Modifying the scanning software with a custom-written macro to address the scanning mirrors in combination with rapid intensity switching by an electro-optic modulator is sufficient to accomplish the acquisition of two-photon striped-illumination patterns on an sCMOS camera. We demonstrate and analyse the resulting resolution improvement by applying different recently published image resolution evaluation procedures to the reconstructed filtered widefield and super-resolved images. This article is part of the Theo Murphy meeting issue 'Super-resolution structured illumination microscopy (part 1)'.
Stichworte
SIM; structured illumination microscopy; super-resolution optical; microscopy; multi-photon fluorescence excitation; laser scanning; fluorescence microscopy
Erscheinungsjahr
2021
Zeitschriftentitel
Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences
Band
379
Ausgabe
2199
Art.-Nr.
20200300
ISSN
1364-503X
eISSN
1471-2962
Page URI
https://pub.uni-bielefeld.de/record/2955094

Zitieren

Pilger C, Pospisil J, Müller M, et al. Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope. Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences . 2021;379(2199): 20200300.
Pilger, C., Pospisil, J., Müller, M., Ruoff, M., Schuette, M., Spiecker, H., & Huser, T. (2021). Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope. Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences , 379(2199), 20200300. https://doi.org/10.1098/rsta.2020.0300
Pilger, C., Pospisil, J., Müller, M., Ruoff, M., Schuette, M., Spiecker, H., and Huser, T. (2021). Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope. Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences 379:20200300.
Pilger, C., et al., 2021. Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope. Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences , 379(2199): 20200300.
C. Pilger, et al., “Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope”, Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences , vol. 379, 2021, : 20200300.
Pilger, C., Pospisil, J., Müller, M., Ruoff, M., Schuette, M., Spiecker, H., Huser, T.: Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope. Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences . 379, : 20200300 (2021).
Pilger, Christian, Pospisil, Jakub, Müller, Marcel, Ruoff, Martin, Schuette, Martin, Spiecker, Heinrich, and Huser, Thomas. “Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope”. Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences 379.2199 (2021): 20200300.

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