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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 cameraThis 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

5x

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

5x

10x

Type

Illumination Optics Lightsheet Z.1 5x/0.1

Illumination 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

50mW

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

 


Related Links
Light Sheet Information from Zeiss

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