Multiplexed confocal microscope with a refraction window scanner and a single-photon avalanche photodiode array detector

Currently, the spinning disk confocal microscope is the most popular fast confocal imaging system. However, these instruments require a high pixel density camera detector, limiting their use in many applications. We have designed a multiplexed confocal scheme that performs raster scanning of a ${32 \times 32}$32×32 foci array in which the output image is stationary after de-scanning. Our system can be coupled to a dense or sparse array detector, making it feasible to apply to rapid time-resolved photon counting, single-photon detection, and some super-resolution techniques.

[1]  Angelo Gulinatti,et al.  High-throughput FCS using an LCOS spatial light modulator and an 8 × 1 SPAD array , 2010, Biomedical optics express.

[2]  Qiyin Fang,et al.  High-speed multifocal array scanning using refractive window tilting. , 2015, Biomedical optics express.

[3]  Juha Kostamovaara,et al.  Fluorescence suppression in Raman spectroscopy using a time-gated CMOS SPAD. , 2013, Optics express.

[4]  Richard Berman,et al.  Spinning-disk confocal microscopy: present technology and future trends. , 2014, Methods in cell biology.

[5]  Jos Joore,et al.  High-throughput compound evaluation on 3D networks of neurons and glia in a microfluidic platform , 2016, Scientific Reports.

[6]  E. Charbon,et al.  Architecture and applications of a high resolution gated SPAD image sensor. , 2014, Optics express.

[7]  Clifford M. Babbey,et al.  Performance comparison between the high‐speed Yokogawa spinning disc confocal system and single‐point scanning confocal systems , 2005, Journal of microscopy.

[8]  Qiyin Fang,et al.  Characterization of SPAD Array for Multifocal High-Content Screening Applications , 2016 .

[9]  Yonghong Shi,et al.  High-content screening identifies kinase inhibitors that overcome venetoclax resistance in activated CLL cells. , 2016, Blood.

[10]  Lei Wang,et al.  The Ustilago maydis repetitive effector Rsp3 blocks the antifungal activity of mannose-binding maize proteins , 2018, Nature Communications.

[11]  Claire M Brown,et al.  Any Way You Slice It-A Comparison of Confocal Microscopy Techniques. , 2015, Journal of biomolecular techniques : JBT.

[12]  Maximilian T. Strauss,et al.  Multiplexed 3D super-resolution imaging of whole cells using spinning disk confocal microscopy and DNA-PAINT , 2017, Nature Communications.

[13]  J Schwider,et al.  Confocal microscopy with a refractive microlens-pinhole array. , 1998, Optics letters.

[14]  Qiyin Fang,et al.  Streak camera crosstalk reduction using a multiple delay optical fiber bundle. , 2012, Optics letters.

[15]  R Richards-Kortum,et al.  Fiber-optic confocal microscopy using a spatial light modulator. , 2000, Optics letters.

[16]  David Stoppa,et al.  Coincidence detection of spatially correlated photon pairs with a monolithic time-resolving detector array. , 2016, Optics express.

[17]  Qiyin Fang,et al.  Cross-talk reduction in a multiplexed synchroscan streak camera with simultaneous calibration. , 2019, Optics express.

[18]  M. Davidson,et al.  An Improved Cerulean Fluorescent Protein with Enhanced Brightness and Reduced Reversible Photoswitching , 2011, PloS one.

[19]  D. Andrews,et al.  Interactions of pro-apoptotic BH3 proteins with anti-apoptotic Bcl-2 family proteins measured in live MCF-7 cells using FLIM FRET , 2012, Cell cycle.

[20]  Shoji Kawahito,et al.  A 10 ps Time-Resolution CMOS Image Sensor With Two-Tap True-CDS Lock-In Pixels for Fluorescence Lifetime Imaging , 2016, IEEE Journal of Solid-State Circuits.