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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68407700%3A21230%2F21%3A00345383" target="_blank" >RIV/68407700:21230/21:00345383 - isvavai.cz</a>

Výsledek na webu

<a href="https://doi.org/10.1098/rsta.2020.0300" target="_blank" >https://doi.org/10.1098/rsta.2020.0300</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1098/rsta.2020.0300" target="_blank" >10.1098/rsta.2020.0300</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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 utilizing these methods is the need for new, complex optical setups. 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 twophoton laser scanning microscopy can readily be exploited for achieving optical super-resolution and contrast enhancement utilizing 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 a 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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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 utilizing these methods is the need for new, complex optical setups. 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 twophoton laser scanning microscopy can readily be exploited for achieving optical super-resolution and contrast enhancement utilizing 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 a 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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10306 - Optics (including laser optics and quantum optics)

Projekt

—

Návaznosti

S - Specificky vyzkum na vysokych skolach

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences

ISSN

1364-503X

e-ISSN

1471-2962

Svazek periodika

379

Číslo periodika v rámci svazku

2199

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

11

Strana od-do

1-11

Kód UT WoS článku

000644937400007

EID výsledku v databázi Scopus

2-s2.0-85105905329