Nikon Ti2E-03 inverted / PRIMO

Nikon Ti2E
Imaging System

Specifications:

  • Timelapse option, motorized stage
  • Perfect Focus System (PFS)
  • Photometrics Moment 12 bit CMOS camera
    (3200 x 2200 px, 50 fps)
  • Andor iXon X3 EMCCD camera
    (1004 x 1002 px, 32 fps)
  • DIC imaging with 10x, 20x, 20x ELWD and 40x objectives
  • Nikon JOBS software: automation of acquisition
  • Heating unit and Gas incubation system to control CO2 and O2 concentration

Installed Objectives:

  • Plan Fluor 4x/0.2 air
  • Plan Fluor 10x/0.3 DIC 1 N1 air PFS
  • Plan Apo λ 20x/0.75 DIC 1 air PFS
  • Plan Apo λ 40x/0.95 DIC air PFS
  • S Plan Fluor ELWD 20x/0.45 air PFS

Emission filters on lower filter wheel (for Acquisition with NIS)

    • DAPI (BS 409, BP 447/60)
    • FITC (BS 495, BP 525/50)
    • TRITC (BS 573, BP 609/54)
    • Cy5 (BS 660, BP 692/40)
    • 5-band filter: DAPI, FITC, TRITC, Cy5 & Cy7

Lumencor LED light source with flexible excitation band selection:

    • Violet (395 nm)
    • Blue (440 nm)
    • Cyan (470 nm)
    • Teal (508 nm)
    • Green (555 nm)
    • Red (640 nm)

Manual Sliders at TL lamphouse

        • GIF (Green Interference Filter to minimize backreflection)
        • ND8A neutralfilter

System manual and additional material is available on SeaFile (login required).

Resources:

To open ND2 files in FIJI: go to Plugins > Bio-Formats > Bio-Formats Plugins Configuration. Under the tab Formats find Nikon ND2 (it is sorted alphabetically) and disable Use chunkmap table.

If this does not work, try enabling the Nikon reader or different combinations of the last 2 check boxes.

 

Alvéole PRIMO 2
Photo Patterning System

Specifications:

  • Alvéole Primo System:
    Digital Mirror Device (DMD) for UV projection
  • Perfect Focus System (PFS)
  • Photometrics Moment 12 bit CMOS camera
  • Alvéole Leonardo software for control
  • Gas incubation system to control O2 concentration, namely for Hydrogel structuration

Compatible Objectives:

  • Plan Fluor 4x/0.2 air
  • Plan Fluor 10x/0.3 DIC 1 N1 air PFS
  • S Plan Fluor ELWD 20x/0.45 air PFS

Filters on upper filter wheel (for PRIMO experiments with Leonardo)

    • Dichroic Mirror (UV: Ex <405 nm/Em >405 nm)

Light sources:

    • PRIMO 2 365 nm LED light sourse
    • Nikon Ti2E Transmitted Light

Possible Applications & Requirements

The Alvéole PRIMO system can be used for different applications, different reagents are required, as follows:

Micropatterning

    • Micropatterning
      – Substrates to be patterned on (glass coverslip/dishes, EM grids etc.)
      – PLPP photo initiator (gel or liquid)
      – Plasma cleaner/Glow discharger
      – PBS 1X
      – MQ water
      – Ethanol (at least 70%)
      – HEPES, 0,1 M 8.3<pH<8.5
      – Poly-L-Lysine 0,01 %
      – Molecules to be patterned (ECM protein, etc.)

Hydrogel Structuration

    • Hydrogel structuring
      – Hydrogel of interest (e.g. 4-Arm-PEG-Acrylate, PEGDA, Matrigel, etc.
      for more information see “Resources”)
      – PLPP photo initiator (liquid)
      – PBS 1X

Spoiler title

    • Microfabrication
      – Spin-coated wavers (SU8, etc.) 
      – Developer for coating
      – basic material for photolithography (chemical hood, hot plate, glass crystallizers, isopropanol, etc.)

System manual and additional material is available on SeaFile (login required).