TY  - JOUR
AB  - 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)'.
DA  - 2021
DO  - 10.1098/rsta.2020.0300
KW  - SIM
KW  - structured illumination microscopy
KW  - super-resolution optical
KW  - microscopy
KW  - multi-photon fluorescence excitation
KW  - laser scanning
KW  - fluorescence microscopy
LA  - eng
IS  - 2199
PY  - 2021
SN  - 1364-503X
T2  - Philosophical Transactions of the Royal Society of London, Series A : Mathematical, Physical and Engineering Sciences
TI  - Super-resolution fluorescence microscopy by line-scanning with an unmodified two-photon microscope
UR  - https://nbn-resolving.org/urn:nbn:de:0070-pub-29550943
Y2  - 2024-11-22T04:03:41
ER  -