Total Internal Reflection Fluorescence (TIRF) Microscope

The TIRF microscope is located in the BSL-2 facility on the first floor. The TIRF set up consists of an Olympus IX71 Inverted research microscope equipped with a motorized stage allowing sample movement in X and Y axis as well as fine focusing in the z axes. The whole set up rests on a Vibration Isolation Stage (TMC, Peabody MA) connected to the compressed air line.

Light Sources

• A 100 W (max output) halogen lamp for Bright field, DIC, Phase Contrast and Trans-illumination.
• X-Cite 120PC short arc lamp (output 120 W) for fluorescence microscopy.
• Blue solid state 30 mW Melles Griot laser (wavelength = 488 nm) for TIRF.

Objectives

• 10x (NA = 0.3) for phase contrast and bright field microscopy.
• 100x (NA =1.45) oil TIRFM objective for TIRF, epi-fluorescence, bright field, DIC and trans-illumination microscopy.

Camera

• HAMAMATSU digital CCD C10600-10B ORCA-R2 16 bit camera. The camera transfers images directly to a PC through IEEE1394b interface. The camera is equipped with high resolution solid state imaging device with a resolution of 1344 (horizontal) x 1024 (vertical) pixels. The camera provides a maximum frame rate of 60 fps.

The set up also includes a Dell Precision T3400 PC (Intel Core 2 Duo CPU-3.16 GHz and 3.25 GB RAM). The camera, motorized stage, arc lamp, and the laser can be controlled using SlideBook 4.2 software (Intelligent Imaging Innovations) installed on the PC. This software allows the user to perform real time as well as time lapse video microscopy and has also built in tools for image analysis.

Prospective users may contact - Prithu Sundd, Ph.D, IB-Ley

For training please contact - Leo Fernandez, Director Imaging Facility