The TruLive3D is a dual-side illumination light sheet that is optimized for fast and gentle imaging of samples ranging from a few micrometers to approximately 200um deep into the tissue over multiple days. The system is optimized for samples in aqueous solution with special U-bottom FEP chambers to match the refractive index of water and is set up for live imaging with CO2 and temperature control. The chambers can also be used to automate imaging multiple samples with light sheet over time. It is equiped with dual Hammatsu Orca Fire Cameras, a changeable dichroic beam splitter for simultaneous imaging, 6-lasers for imaging and a 535nm pulsed ablation laser.

Cartoon view of imaging set up drawn by Kari Herrington:

This system generates large amounts of Data, and it is hooked up directly to our server to help handle data. Please email us to get set up on the server.

Links

ILAB CALENDAR

FPBASE

Capabilities

• Brightfield 
• 6 lasers- all lasers can be used for Photokinetecs/FRAP
• 535nm Photo Ablation laser
• 4- Channel fluorescent imaging
• 2-color fluorescence simultanious 
• Long live cell imaging
• Fast volume with low photo bleaching

• Automated multi-position imaging 

   

Objectives:

• Plan Apo 25x/1.15NA WI

• Exchangeable tubelenses to change magnification

  • 0.5x = 12x 
  • 1.0x = 25x 
  • 1.5x = 37.5x
  • 2.0x = 50x

   

Equipment Details:

• Bruker Luxendo TruLive3D Light Sheet
  • 6 Omicron 40mW Diode lasers with Micron Light Hub Laser Lombiner for light sheet
  • TAG lens for digital sheet scanning
  • Tip/tilt beam feature for destriping
  • Dual/simultaneous imaging
• Orca Fire Cameras x2
• Ibidi Gas incubation Mizer for CO2 and O2 (Currently only set up for CO2)
• 532 Pulsed CryLas 

Light Sources

Brightfield:

• 880 LED

Excitation Wavelengths for Light sheet:

• 405nm
• 488nm
• 515nm
• 561nm
• 642nm
• 685nm

*All imaging lasers can be used for Photo Manipulation/Kinetics/Bleaching mode

Ablation laser

• 532 Pulsed CryLas

Dichroics (Camera Splitting):

• 500LP
• 565LP
• 660LP 

Emission Filters for Short Camera:

• Emission Filters Short Camera
• BP 418-462 = Chroma ET 440/40
• LP 498 = Semrock 488LP
• BP 497-554 = Semrock 525/50
• BP 526-564 = Chroma ET 545/40
• BP 580-627 = Chroma ET 605/50
• BP 610-629 = Semrock 620/14 

Emission filters for Long Camera

• LP 498 = Semrock 488LP
• BP 497-554 = Semrock 525/50
• BP 526-564 = Chroma ET 545/40
• BP 580-627 = Chroma ET 605/50
• BP 610-629 = Semrock 620/14
• BP 655-704  = Semrock Brightline 679/41
• LP 656 =  Chroma 655 LP
• LP 696 = Chroma 700LP
• BP 710-749 = Semrock 730/39