Evanescent Wave Microscopy (1 system)

Nikon TIRF (Tel. 94 782)

Capabilities    TIRF.pdf

  • Observations of biological structures near the plasma membrane (around 100 nm)
  • For fixed samples labelled in fluorescence (green and/or red)
TIRF.jpg

Detailed specifications

Microscope

Nikon Eclipse Ti with PFS

Objectives

10x 0.3 Plan Fluor DIC C / N1  WD 16mm

40x 1.3 Oil Plan Fluor DIC H  WD 0.2mm

63x 1.4 Oil Plan Apo DIC H  WD 0.21mm

100x 1.49 Oil CFI Apochromat TIRF

 

Mounted Filters

1) TIRF Quad Exciter+Dichroic  405/488/561/64
 Cube_TIRF_Dichroic.pdf
2) DAPI 5060 C : ex 377/50  FF409  em 447/60
3) TIRF FITC : ex 488/10  DM502  em 530/43
4) TIRF TRITC : ex 561/10  DM580  em 605/50
5) SRIC or Cy5 4040 C : ex 628/40  FF660  em 692/40  (Please ask the team)

6) DAPI Dichroic ONLY (FRET with filters 2.CFP-3.YFP)

Filter wheel emission

  1. Empty
  2. CFP 470/20
  3. YFP 535/30
  4. DAPI435/40
  5. FITC 525/50
  6. Cy3 605/70
  7. 630/75
  8. Cy5 700/75
  9. Close
  10. Close

Lasers

Nikon laser bench LU-N4

405 nm (100 mW)

488 nm (50 mW)

561 nm (50 mW)

640 nm (100 mW)

Camera

ORCA Fusion BT (Hamamatsu), sCMOS camera

The benefits of sCMOS cameras over existing camera technologies include:

  • Better temporal resolution due to higher acquisition frame rates
  • Larger fields of view due to larger chip sizes
  • Better spatial resolution due to smaller pixel sizes that can more properly digitally sample the images produced by high numerical aperture (NA) objective lenses (Nyquist sampling)
  • Better intensity quantification due to a higher dynamic range