Leica STELLARIS 8 FALCON STED

A laser scanning confocal on an inverted Leica DMI8 motorized microscope stand

Features

• White-light laser for optimal excitation from 440nm-790nm plus 405nm diode. Easy 5+ channel imaging!
• 3D STED super-resolution with 775nm depletion beam for resolution down to 50nm or smaller. NOTE: STED specific information below.
• TauSense modules that use lifetime information for filtering
• FALCON for accurate lifetime measurements.
• Sensitive hybrid detectors are capable of photon counting and enable time-gating to reduce autofluorescence background.
• Live cell stage top incubator. NOTE: Requires 35mm dish or chambered coverslips.
• AFC (Automatic Focus Correction) to prevent focal drift

Acknowledgement

Acknowledgement of NIH grant 1S10OD030300 is required for all publications in which data from this instrument is used.

Location: 234D Taylor Hall

Specifications

Objectives

• HC Plan Fluotar 5x/0.15 WD13.7mm
• HC Plan-Apo 10x/0.45 WD2.0mm
• Plan-Apo 20x/0.8WD 0.55mm
• Plan-Neofluar 40x/1.3 oil WD0.21
• Plan-Apo 63x/1.4 oil (partial list)

Excitation Lines

• 405nm
• 440-790 continuous selection with White Light Laser

Detection

• 3 HyD S detectors
• 2 HyD X detectors
• Transmitted light detector (DIC)

STED specific resources

• STED sample prep for HIC document includes links below and some UNC-specific links.
• Do NOT use AlexaFluor 647; it photobleaches almost instantly in STED mode.
• Leica’s STED sample preparation guide (pdf download), including fluorophore guide
• Abberior’s document on STED sample preparation (pdf download)
• We have had success getting STED resolution in 3 colors with AlexaFluor 594, ATTO 647N and CF680R (Biotium).
• For UNC researchers, the HIC has a small bank of reagents for testing STED imaging on your samples. Please contact Wendy for details.
• Places to buy antibodies with STED fluorophores (weblinks, not exhaustive): PLEASE email Wendy other sources!
  • Milipore-Sigma has some standard secondaries with aberrior dyes
  • Rockland has IgG subtype specific antibodies
  • Creative Labs has camelid nanobodies (smaller, less amplification)
  • Biotium (CF dyes) has standard secondary host-target pairs plus donkey-raised secondaries and secondaries against less-common primary hosts (guinea pig, rat, etc). Lists STED compatible dyes.
  • Jackson ImmunoResearch for AlexaFluor 594
  • ThermoFisher/Molecular Probes for AlexaFluor 594
  • Label your own antibodies with dye kits: Sigma (Aberrior dyes, US), Atto-Tec (Aberrior dyes, EU) and ThermoFisher/Molecular Probes (AlexaFluors)