Enhanced sectioning quality of Zebrafish Z-stacks with CICERO Spinning Disk Confocal at ALM i3S

The Advanced Light Microscopy Scientific Platform at the i3S

The Advanced Light Microscopy (ALM) is a scientific platform of i3S-Instituto de Investigação e Inovação em Saúde in Porto, Portugal. The ALM is dedicated to state-of-the-art optical microscopy applications for biosciences. The applications available include multidimensional imaging of cells and tissues, high-speed live cell microscopy, molecular analysis techniques, and in vivo microscopy. The ALM provides access to multiple advanced light microscopy systems, scientific advisement in experiment planning, consulting, and collaboration in research projects, holding an open-access policy where the resources are available to the scientific community and industry. The ALM is a core facility of the national research infrastructure PPBI-Portuguese Platform of BioImaging and the European research infrastructure Euro-Bioimaging ERIC.

Enhanced sectioning quality of Zebrafish Z-stacks with CICERO Spinning Disk Confocal (CrestOptics)

The ALM scientific platform has already adopted CrestOptics X-Light V3 Spinning Disk and DeepSIM X-Light super-resolution technologies to meet the multiple imaging requests of a Microscopy Facility when it comes to balancing speed, resolution, and phototoxicity on a whole bunch of diverse samples. “At i3S, Zebrafish is used as a physiologically relevant organism to model organ development and disease progression. For that purpose, we consider the CICERO Spinning Disk Confocal from CrestOptics as the easiest route to confocality for enhancing the optical sectioning quality across Zebrafish Z-stacks”, Prof. Paula Sampaio, Head of the ALM said. “I was positively impressed by the ease of installation and operation of the CICERO. I mounted it on my own on a ZEISS Axio Imager Z1 upright microscope with just a little remote support from CrestOptics staff. We also coupled the system with a CELESTA laser source (Lumencor) and a BSI Express sCMOS (Teledyne Photometrics) with 95% QE. With the CICERO Spinning Disk unit we smoothly managed to recycle and upgrade our ZEISS upright frame to a confocal system even with a cost-saving budget”, Prof. Paula Sampaio added. “The CICERO performances in terms of speed and confocal resolution met our expectations: we carried out lots of Zebrafish Z-stack acquisitions with optimal out-of-focus light rejection at a fast rate”, Prof. Paula Sampaio, concluded.

Prof. Paula Sampaio is the head of the Advanced Light Microscopy Scientific Platform, founded by her two decades ago and which she has continued to develop. She was also the founder and national coordinator of PPBI – the Portuguese Platform of BioImaging, currently a node of the Euro-Bioimaging ERIC. She lectures on Bioimaging in Masters and Doctoral Courses at the University of Porto and has organized several international open courses and events on microscopy and image analysis, including the course series on Optical Microscopy Imaging for Biosciences and NEUBIAS training courses.

A. WIDEFIELD 

B. SPINNING DISK CONFOCAL

C.

Figure 1. CICERO Spinning Disk Confocal imaging on a ZEISS Axio Imager Z1 upright microscope. Zebrafish samples are stained with DAPI and Alexa Fluor 488 (neuron marker), then 150 μm Z-stacks are acquired with a ZEISS PlanApo 20x/0.8 objective. Maximum Intensity Projection of CICERO widefield (A) and spinning disk confocal (B) imaging modalities are shown. Scale bar, 100 µm. (C) 3D movie of the spinning disk volumetric acquisition. The Zebrafish sample was prepared by Dr. Ana Filipa Castro, from the Genetics of Cognitive Dysfunction group at i3S Porto (PT) and imaged by Prof. Paula Sampaio.

THE EXPERT’S VOICE ON CRESTOPTICS TECHNOLOGY

“We consider the CICERO Spinning Disk Confocal from CrestOptics the easiest route to confocality for enhancing the optical sectioning quality across the Zebrafish Z-stacks. It is very easy to install and operate on our ZEISS Axio Imager Z1 upright microscope.“

Prof. Paula Sampaio, Head of the Advanced Light Microscopy Scientific Platform, i3S Porto (Portugal).

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