Affiliated with Université Laval & CERVO Research Centre

Resolution enhancement in laser scanning microscopy with deconvolution switching laser modes (D-SLAM).

Publication Type:

Journal Article

Source:

Opt Express, Volume 26, Issue 19, p.24881-24903 (2018)

Abstract:

<p>Laser scanning microscopy is limited in lateral resolution by the diffraction of light. Superresolution methods have been developed since the 90s to overcome this limitation. However superresolution is generally achieved at the expense of a greater complexity (high power lasers, very long acquisition times, specific fluorophores) and limitations on the observable samples. In this paper we propose a method to improve the resolution of confocal microscopy by combining different laser modes and deconvolution. Two images of the same field are acquired with the confocal microscope using different laser modes and used as inputs to a deconvolution algorithm. The two laser modes have different Point Spread Functions and thus provide complementary information leading to an image with enhanced resolution compared to using a single confocal image as input to the same deconvolution algorithm. By changing the laser modes to Bessel-Gauss beams we were able to further improve the efficiency of the deconvolution algorithm and obtain images with a residual Point Spread Function having a width of 0.14 λ (72 nm at a wavelength of 532 nm). This method only requires a laser scanning microscope and is not dependent on certain specific properties of fluorescent proteins. The proposed method requires only a few add-ons to classical confocal or two-photon microscopes and can easily be retrofitted into an existing commercial laser scanning microscope.</p>