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)'.
- Klíčová slova
- SIM, laser scanning fluorescence microscopy, multi-photon fluorescence excitation, structured illumination microscopy, super-resolution optical microscopy,
- MeSH
- algoritmy MeSH
- Convallaria ultrastruktura MeSH
- ledviny ultrastruktura MeSH
- mikroskopie fluorescenční multifotonová přístrojové vybavení metody statistika a číselné údaje MeSH
- myši MeSH
- optické jevy MeSH
- optické prostředky MeSH
- počítačové zpracování obrazu metody statistika a číselné údaje MeSH
- software MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
