Three-dimensional image cytometer based on widefield structured light microscopy and high-speed remote depth scanning

• Main Authors: Choi, Heejin (Contributor), Wadduwage, Dushan N. (Author), Tu, Ting Yuan (Author), Matsudaira, Paul T. (Author), So, Peter T. C. (Contributor)
• Other Authors: Massachusetts Institute of Technology. Institute for Medical Engineering & Science (Contributor), Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Mechanical Engineering (Contributor), Massachusetts Institute of Technology. Laser Biomedical Research Center (Contributor), Singapore-MIT Alliance in Research and Technology (SMART) (Contributor)
• Format: Article
• Language: English
• Published: Wiley Blackwell, 2015-07-13T15:05:24Z.
• Subjects: Article
• Online Access: Get fulltext

A high throughput 3D image cytometer have been developed that improves imaging speed by an order of magnitude over current technologies. This imaging speed improvement was realized by combining several key components. First, a depth-resolved image can be rapidly generated using a structured light reconstruction algorithm that requires only two wide field images, one with uniform illumination and the other with structured illumination. Second, depth scanning is implemented using the high speed remote depth scanning. Finally, the large field of view, high NA objective lens and the high pixelation, high frame rate sCMOS camera enable high resolution, high sensitivity imaging of a large cell population. This system can image at 800 cell/sec in 3D at submicron resolution corresponding to imaging 1 million cells in 20 min. The statistical accuracy of this instrument is verified by quantitatively measuring rare cell populations with ratio ranging from 1:1 to 1:10[superscript 5].
National Institutes of Health (U.S.) (Grant 9P41EB015871-26A1)
National Institutes of Health (U.S.) (Grant 5R01EY017656-02)
National Institutes of Health (U.S.) (Grant 5R01 NS051320)
National Institutes of Health (U.S.) (Grant 4R44EB012415-02)
National Science Foundation (U.S.) (Grant CBET-0939511)
Singapore-MIT Alliance for Research and Technology
MIT Skoltech Initiative
Hamamatsu Corporation
David H. Koch Institute for Integrative Cancer Research at MIT (Bridge Project Initiative)