DEMO: Nikon SIM-TIRF, October 4th-20th 2021

Posted onAuthorGabriel Krens

The Bioimaging Facility at IST Austria is hosting a demonstration of a Nikon N-SIM system from October 4th – 20th, 2021.

The N-SIM S Super Resolution Microscope is a unique high-speed structured illumination system that achieves acquisition speeds of up to 15 fps, enabling fast biological processes to be captured at twice the spatial resolution of conventional light microscopes (up to 115nm in XY). Combining the N-SIM S and a confocal microscope gives you the flexibility to select a location in the confocal image and switch to super-resolution to view the desired part of the location in minute detail. (source)

 

 

Technical details of the Demo system

  • Microscope:
    • Ti2-E Inverted Microscope with motorized stage, Z-Piezo Insert and Perfect Focus System
  • Lasers:
    • 405 nm/445 nm/488 nm/561 nm/647 nm
  • Objectives:
    • CFI SR HP Apochromat TIRF 100XAC Oil (NA 1.49)
      • includes an auto correction collar
    • CFI Plan Apochromat Lambda 60XC (NA 0.95)
    • CFI Plan Apochromat Lambda 40XC (NA 0.95)
  • Cameras:
    • 2 x Hamamatsu Orca Fusion sCMOS cameras (Hamamatsu Photonics K.K.)
    • Hamamatsu Gemini-2C Beamsplitter
  • Incubation System
  • SIM Imaging modes
    • TIRF-SIM
    • 2D-SIM
    • 3D-SIM (Reconstruction methods: slice, stack)
  • Multi-color imaging
  • Simultaneous multi-color imaging of two colors
  • Image acquisition time Up to 15 fps (TIRF-SIM/2D-SIM, 2 ms exposure time)
  • Lateral resolution (FWHM of beads in xy):
    • 115 nm in 3D-SIM mode,
    • 86 nm in TIRF-SIM mode
  • Axial resolution (FWHM of beads in z):
    • 269 nm in 3D-SIM mode

 

Specifications summary:

  • Lateral resolution (FWHM of beads in xy) 115 nm*1 in 3D-SIM mode, 86 nm*2 in TIRF-SIM mode
    Axial resolution (FWHM of beads in z) 269 nm*1 in 3D-SIM mode
    Image acquisition time Up to 15 fps (TIRF-SIM/2D-SIM, 2 msec exposure time)
    Reconstructed image size 1024 x 1024 pixels, 2048 x 2048 pixels

Demo-test sessions:

We offer morning and afternoon slots starting from October 4th to 8th (week 1), October 11th to 15th (week 2) and from October 18th to 20th (week 3). The N-SIM system  will stay at IST until the 20th of October in our room in the multi-purpose hall. We will have a super resolution application specialist from Nikon on site from October 4th to October 8th, 2021. In the second Demo week from 11th to 15th of October 2021, the system will be operated by BIF members. External scientists are welcome to try the system.

1st week Mo, October 4th Tue, October 5th Wed, October 6th Thu, October 7th Fri, October 8th
morning 9 to 12 am BIF   C. Gianinni; grad school NIKON BIF
afternoon 1 to 4 pm BIF A.Johnson; Friml group M.Javoor; Schur group BIF  
2nd week Mo, October 11th Tue, October 12th Wed, October 13th Thu, October 14th Fri, October 15th
morning 9 to 12 am NIKON tech NIKON tech      
afternoon 1 to 4 pm NIKON tech NIKON tech N.Canigova; Sixt group   Marko Kojic, Loose grp (cy3 & cy5 on lipid bilayers)
3rd week Mo, October 18th Tue, October 19th Wed, October 20th
morning 9 to 12 am Y.Vallis; De Bono group Uni Wien (Köhler-grp)  
afternoon 1 to 4 pm Suyash NAIK; Heisenberg grp GFP & mCherry Uni Wien (Köhler-grp) Philipp Radler, Loose grp

Requests for demo-slots:

Please provide the following information to bioimaging@ist.ac.at:

  1. Preferred time slot
  2. short description of your sample: fluorophores, module, aim

 

Sample preparation

To prepare/mount your samples please only use 1.5H precision glass coverslips (170 µm +/- 5 µm) (see ref. 1-2 for examples) or high precision (#1.5H) glass bottom dishes (see ref. 3):

      1. Thorlabs 1.5H precision coverslips
      2. Marienfeld 1.5H precision coverslips
      3. Mattek high precision (1.5H) glass bottom dishes

Location: Multi Purose Hall  IST Austria Campus, 3400 Kloserneuburg

Information page: Nikon Ti2-LAPP TIRF system

campus map