Location:
I04.02.031
System description and specification
The Olympus cellTIRF-4Line system is an ultra-sensitive, simultaneous multicolor Total Internal Reflection (TIRF) microscope using four laser channels with independent beam paths. Individual motorized angle control for each laser ensures equal evanescent penetration (±1 nm), which enables the user to produce high-contrast images with minimal background noise for cell surface and single molecule studies.
Since the angle of each laser light source is independently motorized, their penetration depths can be synchronized across each wavelength using cellSens software. This also provides key data about the penetration depths that are possible with each laser/objective pair.
- Olympus IX83 inverted microscope equipped with a Cell^TIRF-4-line module
- Multicolor TIRF with independent angle control
- Motorized stage for multiple position imaging
- Incubator and CO2 supply for live cell imaging
Scheme of the Cell^TIRF-4-line system
light sources:
- CW Diode Lasers, laser lines 405 nm, 488 nm, 561 nm, 640 nm
- Transmitted light
filter sets:
- Quadfiltercube: zt405-488-561-640rpc from Chroma (USA) and FF01-446/523/600/677-25 from Laser2000 (UK)
- GFP filter cube: LP491; BP 525/45
camera:
- Hammamatsu ImagEM X2 EM-CCD: C9100-13, (512×512 pixels, pixel size = 16µm)
software:
cellSens Dimension 2.2
dual channel splitting optics:
W-VIEW GEMINI image splitting optics from Hammamatsu (Japan) provide one pair of dual-wavelength images separated by a dichroic mirror onto a single camera.
Filter options are: Green (524/27) /Red (617/73); Green (525/50) /FarRed (700/75); Red (579/34)/FarRed(700/75)
Dichroic mirrors: T580; T640
Objectives
- Olympus UAPON OTIRF 100x /1.49 OIL for TIRF imaging
- Olympus Uplan SApo 100x/1.40 OIL
- UAPON 0340 40xOIL available for brightfield and widefield fluorescence modes only !!!
- TIRF Tool 0.5x for alignment purposes, BIF Only !!!
System manual and additional material is available on SeaFile (login required).
contact persons: