Configuration of the RARAF microbeam endstation.
Schematics of immersion Mirau interferometric attachment.
Components of SIMI: The light from a light source (LS) passes through a collimator lens (CL), a linear polarizer (P), and a band-pass filter (F) is reflected by a 50/50 beam-splitter (BS) and directed to the objective (O) with mounted SIMI interferometer (M). The interferometric attachment fits into the microbeam Petri dish (D). A λ/8 waveplate, placed in the test arm of the interferometer, introduces a quarter-wave phase shift to one component of the test beam (inset). Successive application of a polarization beam-splitter (PBS), that follows a tube lens (TL), enables separation of two interference patterns with different phase that are recorded on camera sensors (C1, C2). The arrows denote the directions of polarization.
Design drawing of immersion Mirau interferometric attachment: 1—attachment body, 2—optics holders, 3—beam-splitter, 4—mirror, 5—adjustment screw, and 6—spring.
Modifications of the camera adaptor of the SIMI microscope module: (a) adaptor for one camera with a polarization beam-displacer fitted inside; (b) adaptor for two cameras containing a polarization beam-splitter.
SIMI images of live cells. (a) 3T3 cells were plated on mica substrates, illumination wavelength 540 nm; (b) WI 38 cells were plated on glass slides; illumination wavelength 605 nm. In both cases the cells were cultured in cell growth medium and imaged in PBS; to create each image, two interference patterns were formed on a single camera sensor. (c) HT 1080 cells plated on glass slides; illumination wavelength 540 nm. The cells were imaged in clear medium using two camera sensors.
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