Lightsheet at CIB U. Major.

The Light-Sheet Fluorescence Microscopy (LSFM) is ideal for fast and smooth imaging of living organisms in long developmental stages. Also, this microscopy is widely used to obtain deep images of fixed tissues that are naturally transparent or pre-clarified. Rapid imaging, with low phototoxicity and photo-bleaching, is due to Single Plane Illumination Microscopy (SPIM) system. In this, the illumination and detection are performed separately with dedicated targets, where the detection is orthogonal to the sample illumination. This optical configuration delivers excellent contrast in the images, avoiding out-of-focus fluorescence typically obtained with other systems using a low power intensity in the excitation laser, thus protecting the sample from damage.

Our ZEISS Lightsheet 7 microscope enables imaging with cellular resolution of both living specimens and large specimens up to 2 cm. This equipment is also compatible with expansion microscopy, increasing the optical capabilities of our device. In addition, for clarified samples, the equipment has specialized optics, immersion chambers and special sample holders, which allow adaptation of our system to use various types of samples, processed in various clarification protocols using different Refractive Indices in their mounting media.

Our system is equipped with the following features:

• Single plane illumination microscopy (SPIM) Lighsheet microscope Zeiss 7, with dual side illumination, ultra fast pivoting and 4 axis sample position device.
• Two 20x detection objectives with IR correction ring (Water Plan-APO 20x/0.1 and PN 20x/0.1 Corr nd=1.45) for water and clarity immersion media respectively. One 5x detection objective (EC Plan-NEOFLUAR 5x/0.16) for 1.33-1.58 IR medium.
• Two sets of two illumination objectives (10x and 5x) with IR correction ring from 1.33 to 1.58.
• 4 solid state laser excitation channels (405,488, 561, 638).
• 1 laser bloquer filter 405/488/561/638
• 3 doble filter sets: GFP-Cy3, GFP-mCherry, GFP-DRAQ5, Cy3-DRAQ5
• 1 chamber for 20x water immersion objective (IR 1.33); 1 chamber for 20X immersion media objective (IR  1.35-158  ) and 1 chamber fully close for 5x objectives (IR 1.33-1.58)
• 1 Module for temperature control for in vivo sample record