Cell TIRF-4 Line System

Location

RL B.717CB

Capabilities

  • Total Internal Reflection Fluorescence (TIRF) and Highly-Inclined Laminated Optical sheet (HILO) imaging
  • Variable TIRF depth
  • Temperature, humidity, CO2 control
  • 4 channels with individual beam paths with FRAP optics for first line
  • Beam splitter for simultaneous 2 color imaging

Cost

Internal user $29/hour, internal service fee $50/hour

Detailed Specifications

  • Olympus IX83 Fully Motorized Inverted Microscope
  • Zero Drift Compensator (IX3-ZDC2) laser-based autofocus
  • Ultrasonic automated stage and Multiple Area Time Lapse Stage Control Software, as well as Well Plate Navigator Tool for multi-area time-lapse and multi-area mosaic image stitching
  • Tokai hit stage-top live-cell incubation chamber

Light Sources

  • Laser diode 405 nm (100 mW), 488 nm (100 mW), 561 nm (100 mW) (DPSS), 640 nm (100 mW)
  • X-CITE 120 LED-Based Transmitted and Epifluorescence Illumination

Conventional Fluorescence Filters

  • Dapi
  • Green
  • Red

Objectives

Air objectives

  • UPLFLN4xPH; U PLAN FLUORITE 4x phase, NA 0.13, WD 17 mm
  • UPLFLN10x2PH; U PLAN FL 10x phase, NA 0.3, WD 10 mm
  • UCPLFLN20xPH; C PLAN FL 20x phase, Correction collar, NA 0.7, WD 0.8-1.8 mm
  • LUCPLFLN40xPH; LWD U PLAN FL 40x phase, Correction collar, NA 0.6, WD 2.7-4.0

Oil objective for TIRF and HILO microscopy

  • UAPON 100 x OTIRF; UAPO 100 x TIRF , NA 1.49, WD 0.1 mm

Dichroic(s)

  • TRF49901-OL3; ET-405 nm LASER TIRF SET
  • TRF49904-OL3; ET-488 nm LASER TIRF SET
  • TRF49904-OL3; ET-488 nm LASER TIRF SET
  • TRF49909-OL3; ET-561 nm LASER TIRF SET
  • TRF49914-OL3; ET-640-647 nm LASER TIRF SET
  • TRF89901-OL3; ET-405/488/561/640 nm Laser Quad TIRF
  • W-View Gemini optical splitter for simultaneous smFRET detection

Detectors

  • IXON ultra 897 EMCCD, 56 FPS, 512 x 512, 16 µm pixel size.

Computer and Software

  • HP Windows 764 bit Confocal Workstation and 30” Monitor 2 x 1 TB HD, RAID 1, 32GB DDR3
  • Olympus CellSens Integrated Software for Automated Multi-Dimensional Acquisition, Multi-Area Stitching, and Well Plate Scanning. Includes analysis for FRET and Real-Time Intensity Analysis.
  • Software controlled laser incidence angle and penetration depth