A light sheet confocal microscope for image cytometry with a variable linear slit detector
by Hutcheson, J. A.; Khan, F. Z.; Powless, A. J.; Benson, D.; Hunter, C.; Fritsch, I.; Muldoon, T. J.
We present a light sheet confocal microscope (LSCM) capable of high-resolution imaging of cell suspensions in a microfluidic environment. In lieu of conventional pressure-driven flow or mechanical translation of the samples, we have employed a novel method of fluid transport, redox-magnetohydrodynamics (redox-MHD). This method achieves fluid motion by inducing a small current into the suspension in the presence of a magnetic field via electrodes patterned onto a silicon chip. This on-chip transportation requires no moving parts, and is coupled to the remainder of the imaging system. The microscopy system comprises a 450 nm diode 20 mW laser coupled to a single mode fiber and a cylindrical lens that converges the light sheet into the back aperture of a 10x, 0.3 NA objective lens in an epi-illumination configuration. The emission pathway contains a 150 mm tube lens that focuses the light onto the linear sensor at the conjugate image plane. The linear sensor (ELiiXA+ 8k/4k) has three lateral binning modes which enables variable detection aperture widths between 5, 10, or 20 m, which can be used to vary axial resolution. We have demonstrated redox-MHD-enabled light sheet microscopy in suspension of fluorescent polystyrene beads. This approach has potential as a high-throughput image cytometer with myriad cellular diagnostic applications.
- Conference
- High-Speed Biomedical Imaging and Spectroscopy: Toward Big Data Instrumentation and Management
- Year
- 2016
- URL
- https://dx.doi.org/10.1117/12.2211164
- ISBN/ISSN
- 1996-756X; 0277-786X; 9781628419542
- DOI
- 10.1117/12.2211164