Description:

Light-sheet  fluorescence microscopy (LSFM) is known also as selective/single plane illumination microscopy (SPIM). In this technique, the sample is illuminated by a thin sheet of excitation light (usually a few hundred nanometers to a few micrometers) which penetrates the specimen perpendicular to the axis of observation. Consequently, the entire plane of focus can be imaged simultaneously, generating an inherent optical section by exciting only fluorescence from the in-focus plane. Then, the emission light is collected by a  sCMOS camera. This allows high speed optical sectioning imaging of large 3D samples (extremely faster than any other microscopy techniques) at subcellular resolution with high sensitivity and minimal photo-toxicity. Also the good sectioning capability reduces the background signal and thus creates images with higher contrast, comparable to confocal microscopy.

The sample inside the LSFM can be rotated and tilted, thus enabling multi-view imaging of the sample with improved axial resolution.

Dual-sided beam illumination with two aligned objectives complete the 3D imaging of the sample with high resolution.

The exceptional stability of Lightsheet 7 lets you observe living samples over extended periods of time – even days – with less phototoxicity than ever before. Light sheet 7 microscope is designed to image very large optically cleared specimens with subcellular resolution. The Lightsheet 7 have dedicated optics, sample chambers and sample holders to accurately adjust to the refractive index of your chosen clearing method, and then image your large samples, even whole mouse brains.

Features:

• Dual sided illumination
• 2 sCMOS cameras for acquisition of 2 channels simultaneously
• Full incubation with control over temperature and CO2
• Water Chamber
• Clearing chamber for using with 5x foc objective for cleared sample

• Detection objectives:

 

Magnitude	5X (Clearing)	5X	20X	40X
Type	EC Plan-APO 5×/0.16 Foc	EC Plan – NEOFLUAR 5x/0,16	W Plan-APO 20x/1,0 DIC	W Plan – APO 40x/1,0 DIC
NA	0.16	0.16	1.0	1.0
Immersion	Dry	Dry	Water	Water

 

Sheet lens:

 

Magnitude	5x (Clearing)	5x	10X
Type	illumination objective 10×/0.2 foc	Illumation Optics Lightsheet Z.7 5x/0.1	Illumation Optics Lightsheet Z.7 10x/0.2
NA	0.2	0.1	0.2
Immersion	Dry	Dry	Dry
Use with Detection Objectives	5x/0.16 foc	5x	20x, 40x

 

Solid State Lasers:

 

	1	2	3	4
Type	UV	Blue	Green	Red
Wavelength	405nm	488nm,	561nm	638nm
Power	20 mW	50 mW	50 mW	75 mW

 

Epi-Fluorescence Filters:

 

Filter Set #	9310	9313	9316	9317	9322
Fluorophore	DAPI – GFP	CFP – YFP	GFP – Cy3	GFP – mCherry	Cy3 – DRAQ5
Short Emission	420-470	460-500	505-545	505-545	570-640
Beam Splitter	LP 490	LP 510	LP 560	LP 560	LP 650
Long Emission	505-545	525-565	570-640	LP 585	LP 660

 

Applications:

 

Selected application fields of Lightsheet 7:

• Imaging a large (tile) optically cleared specimens with subcellular resolution.
• Morphogenesis and embryogenesis:  Fluorescence imaging of spatial-temporal patterns within cells during embryogenesis of model organisms such as Zebrafish and Drosophila.
• Organogenesis and cell dynamics: Fast imaging of cellular dynamics, such as cell migration, cardiac development, blood flow, vascular development, neuro-development or calcium imaging.
• 3D cell cultures: Live imaging of 3D cell cultures, spheroids and cysts, tissue and organotypic cultures, 3D matrices of cells in polymers and scaffolds, 3D imaging of embryonic bodies and more. Images can be analyzed for cell migration, expression patterns and cell proliferation, cell localization, cell morphology and development of 3D cell cultures.
• Structural imaging of fluorescently labeled living specimens: Imaging detailed volume of fixed and live specimens, for example zebrafish, C-elegance, Arabidopsis root.
• Imaging of marine organisms: Fluorescence imaging of marine organisms such as ciona, squid, plankton and flatworms.
• Single cell imaging: Single cells primarily benefit from the low bleaching and can be studied for morphology, motility and biophysics of microtubule dynamics.

 

Analysis:

Additional computation system is available for the analysis of the Light Sheet data

 

 

Releted Links

 

Light Sheet Information from Zeiss