Leica TCS SP8 DIVE CS (DeepSee dual, FALCON)

The Leica TCS SP8 DIVE CS is a unique microscope due to its large working distance and ability to handle a wide variety of multi-photon imaging applications, such as deep live imaging, capture calcium dynamics (resonance scanner and fast stage), cleared tissue imaging (special objectives), and complex multi-color acquisitions, such as (b)rainbow, due to the option to dynamically tune the detection windows. In addition, the system allows for life-time imaging in MP, and time-gating of undesired signal (post-processing).

 

Location: room i04.U1.018

Imaging modalities/features:

  • Multi-photon (680-1300 nm + 1045) – external 4-tune scan head
  • Confocal (405nm, 488nm, 552nm, 638nm) – internal scan head
  • Second Harmonics Generation (and Third Harmonics Generation)
  • Life-time imaging (FALCON, time gated acquisition )
  • Real-time deconvolution (Lightning)
  • Spectral unmixing
  • Excitation Λ / emission λ wavelength scans (in multi-photon)

Specification:

  • Leica DMi8 CS fixed stage upright  microscope for larger specimen imaging, with cascaded optical table
  • TCS SP8 FALCON, with resonance scanner (16000 Hz – bidirectional)
  •  Scientifica  Motorised Moveable Base Plate (MMBP) Scanning stage (step-sice 0.1 μm, range 50mm, max load: 30 kg)
  • Super Z Galvo Stage Type H (Maximal travel range in z: 1500 μm Minimal step size: 20 nm)
  • Trigger box

Light sources:

  • Cool LED fluorescence lamp (visual inspection)
  • VIS-Laser: 405nm (50 mW), 488nm (20mW), 552nm (20mW), 638nm (30mW) diodes
  • Multi-photon: IR Laser Spectra Physics InSight DeepSee (680-1300 nm + Dual 1045)

Filter for visual examination:

  • Triple band-pass → CFP/YFP/RFP: BP 436/20, BP 503/21, BP 575/25 Emission: BP 470/25, BP 535/30, BP 630/70

Objectives:

  • Default: Objective turrent (27mm):
    1. HC PL Fluotar 10x/0.30, WD 2.2mm – no immersion (VIS only)
    2. HC PL APO 40x/1.10 W CORR CS2, WD 0.65mm – water immersion, Correction collar for use with 0.14­ – 0.18 mm coverglass. (470-1200 nm)
    3. HC PL APO 63x/1.40 OIL CS Plan, WD 0.14mm
    4. HC L Fluotar 16x / 0.60 MI (# 155065), WD 2.5 mm – ne 1.33-1.52 (water, silicon- or glycerol immersion – (Correction collar for use with 0.0­ – 0.17 mm coverglass) + BABB cleared samples
    5. Single objective: HC FUOTAR L 25x / 0.95 W (# 15506374), WD=2.5 mm, wide angle (41°)
    6. Empty
  • Additional high-end rental objectives:
    1. Default  objective – Single objective holder (32mm):
      1. HC IRAPO L 25x/1.00 W mot IRAPO, WD 0.62mm – water, motoCORR (470-1200 nm) – €€€ extremely high-value objective

    2. Single objective holder (32mm):
      1. HC IRAPO L 16x/0.8 W mot VISIR, WD 8.15mm – water, motoCORR (470-1200 nm) – €€€ extremely high-value objective

Internal scan head detectors: confocal- mode

  1. PMT
  2. HyD SP GaAsP­ Detector
  3. HyD SP GaAsP­ Detector

External (4-tune) NDD detectors: multi-photon mode

  1. PMT
  2. HyD2 SMD: specially selected for Falcon FLIM
  3. HyD2 SMD: specially selected for Falcon FLIM
  4. PMT

Detection Filters:

  • ­NF 445
  • NF 488
  • ­NF 514
  • NF 458/514
  • NF 488/561/633 ­
  • NF 594
  • NF 445/594

Modules:

  • LAS X Dye Finder
  • LAS X 3D Visualization
  • LAS X Live Data Mode
  • LAS X FALCON + Phasor FLIM online
  • LAS X LIGHTNING Expert: (deconvolution)

Computer (HPZ840 Workstation – Win 10 professional – 64 bit):

  • 2x Intel 4­core Xeon E5-­2637 V3 3.5 GHz
  • 128 GByte RAM
  • NVIDIA Quadro K4200 4GB high performance GPU ­
  • 256 GB SATA SSD
  • 2x 1TB SSD RAID 0
  • 5x 3TByte SATA hard disk drive, RAID5
  • 10 Gb network card

Manual

Detailed description and manual

How to use the FALCON FLIM detector.


Contact: bioimaging@ist.ac.at

Request a training: training request form