Light Sheet Florescent Microscope

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.

Features:

  • Dual sided illumination
  • 2 sCMOS cameras for acquisition of 2 channels simultaneously
  • Full incubation with control over temperature and CO2
  • Detection objectives:

 

Magnitude X5 20X 40X
Type EC Plan –

NEOFLUAR

5x/0,16

W Plan-APO

20x/1,0 DIC

W Plan – APO

40x/1,0 DIC

NA 0.16 1.0 1.0
Immersion Dry  

Water

Water

 

Sheet lens:

 

Magnitude X5 10X
Type  Illumation Optics

Lightsheet Z.1 5x/0.1

Illumation Optics

Lightsheet Z.1 10x/0.2

NA 0.1 0.2
Immersion Dry Dry
Use with

objectives

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 LSFM:

  • 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