Confocal microscope LSM 900 with Airyscan 2

Comparing the confocal image (left) with Airyscan SR (middle) and Airyscan Joint Deconvolution (right). HeLa cell, 4x expanded and labelled with acetylated alpha tubulin (green).

Location: Unità di Biologia Cellulare e dello Sviluppo, Dept. of Biology, SS Dell' Abetone e del Brennero 4, 56123 Pisa (PI)

Lab head: Prof. Ranieri Bizzarri

Technical assistance: Dott. Sofia Cristiani; Dott. Sara Migliarini; Dott. Luca D’Alessandro

Book the lab

Laboratory rules


Price List:
> Membri UNIPI con accesso autonomo dopo training: 10 €/h;
> Membri UNIPI con operatore: 20 €/h;
> Utilizzo concomitante di strumenti di filiera (Operetta, SEM, TEM): costo da concordare con altri responsabili di strumentazione Non-Ordinario (sempre con operatore);
> Altre Università e Enti di ricerca: 30 €/h
> Aziende: 40€/h
È possibile riservare un monte ore, sino a 250/anno, previo preventivo pagamento al CISUP. Mettersi in contatto con ranieri.bizzarri@unipi.it
 
The Zeiss LSM 900 Airyscan 2 is a fluorescence microscope dedicated to all experimentalists which target functional imaging of biomolecules in living (and fixed) samples with a higher resolution compared conventional confocal systems, in order to image 120-150 nm cell substructures and viruses.

From the technical standpoint, the Zeiss LSM 900 Airyscan 2 combines a highly versatile confocal laser scanning microscope (CLSM) with a detector operating in Image Scanning Microscopy Mode, which is able to return images with 1.7x improved resolution (120 nm on xy plane, 350 nm along the z-axis) as compared to conventional CLSMs. This comes without compromising on Signal to Noise ratio, which is 4x-8x higher as compared to conventional CLSM at nearly same speed. The instrument has spectral imaging capabilities, i.e. detection can be effectively tailored to the spectral features of fluorochromes in the sample.

Operating Modes
Our confocal microscope enables different operating modes:

> Conventional confocal laser scanning mode: three spectral detectors can be activated for (i) concomitant imaging of up to three spectrally distinct fluorophores, and (ii) temporally sequential acquisitions with no limit on the number of fluorophores with different spectral characteristics that can be acquired. Max frame rate/s @512×512 pixel: 6.1;

> Airyscan mode: the airyscan spectral detector can be activated for temporally sequential acquisitions with no limit on the number of fluorophores with different spectral characteristics that can be acquired. Max frame rate/s @512×512 pixel: 4.7 (Airyscan 1x, max resolution: 120 nm), 25 (Airyscan 4x, max resolution: 140 nm);

Both imaging modes can be interleaved and carried out on living samples such as cells by using a thermostated chamber with regulated air flow and CO2 composition.

Recent applications

> Super-resolution imaging of chromatin remodelling in cells;
> Correlative imaging of intracellular nanostructures;
> Diffusion maps of intracellular molecules;
> Super-resolution analysis of virus-cell interactions;
> Super-resolved colocalization imaging of proteins;
> Super-resolution imaging of bacterial biofilms.

Key benefits

> Super-resolution functional imaging of living specimens;
> Spectral acquisition allows for the flexible selection of fluorescent reporters and multiplex imaging;
> Use of airyscan detector for cutting-edge imaging schemes of intracellular diffusion;
> Correlative Light and Electron Microscopy (in combination with SEM).

Off-line image analysis capabilities

A repository of image analysis software (based on the free platform Fiji bundled with custom made plugins) and didactic materials is available to help experimentalists to obtain the maximum information from their acquisitions.

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