SmartSPIM

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Light sheet imaging system optimized for resolution, speed, and flexibility for large samples

SmartSPIM is an advanced light sheet microscope capable of imaging large tissue samples with high speed and resolution. The system was designed from the ground up, with no limitation based on commercially available microscope bodies. The unique illumination arm of the SmartSPIM creates a flat-top illumination profile laterally, and a focused beam axially. To achieve homogeneous axial point spread functions (PSFz) across the entire FOV, the illumination optics scan the beam axially while synchronized to the camera’s rolling shutter detection.

Features and Highlights

  • Capable of imaging a wide range of sample sizes
  • High axial resolution with many different objectives
  • Uniform axial resolution across the entire sample
  • High speed volumetric imaging (mouse hemisphere in less than 15 minutes or a whole mouse brain in less than 25 minutes at 1.75 µm x 1.75 µm x 4 µm voxel size)
  • Ability to use custom and commercially available objectives
  • Custom illumination optics Zemax designed and optimized for achromatic performance and across all points in the sweeping field of view.
  • Detection optics Zemax designed and optimized optimized for all current large format objectives and camera sensors
  • Focus compensation for RI mismatches and chromatic focal shifts
  • Compatible with both aqueous and organic clearing methods

Adult mouse hemisphere imaging acquired with SmartSPIM (8x downsampled from original movie)

Entire hemisphere was stained in 1 day with SmartLabel using SYTO 16 green fluorescent nucleic acid stain and imaged using SmartSPIM with a custom LifeCanvas 3.6x objective. Acquisition time was less than 1 hour. The high and uniform axial resolution allows for clear delineation of cell bodies in XY and importantly in Z.

Specifications

LIGHT SHEET FORMATION

Dynamically axially swept light sheet

ILLUMINATION OPTICS

Custom broadband chromatically corrected optics ( NA = 0.20 )

(higher NA illumination available as a custom item)

SPECIMEN SIZE (MAX)

2 cm x 3.5 cm x 1.2 cm (standard)

3 cm x 5.5 cm x 1.2 cm (custom)

DETECTION OPTICS

Infinity corrected (180 mm EFL tube lens)

LATERAL SAMPLING

3.6x = 1.8 µm/pix; 10x = 0.64 µm/pix; 15x = 0.41 µm/pix

AXIAL RESOLUTION*

3.6x: NA 0.2, PSFz = 3.2-4.0 µm

10x: NA 0.6, PSFz = 1.2-2.2 µm

15x: NA 0.4, PSFz = 1.4-2.4 µm

FIELD OF VIEW (FOV)

3.6x = 3650 µm ; 10x = 1300 µm; 15x = 850 µm 

IMAGING SPEED

20 FPS during z-stack acquisition

(imaging speed can be reduced for increased resolution)

CAMERA

2048 x 2048 sCMOS with rolling shutter – synchronized to sweeping light sheet for confocal detection, high homogeneity, and improved resolution

LASER LINES

Up to 6 in a single combiner (wavelengths from 488 – 785 nm)

*Due to the formation of the light sheet, axial resolution is dependent on imaging rate. The high end of the range is representative of our standard fast acquisition rate.

SmartSPIM image of fluorescent protein expression in a subset of neurons. High-resolution axial spacing (4 µm) enables seamless display of original XY-dimensional data in orthogonal directions (XZ & YZ), providing for accurate identification of each cell body.