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PISA: versatile microscope for 3D single molecule light sheet imaging

Taniguchi, Y.; Nishimura, K.; Yoshida, Y.; Kim, S.; Kamarulzaman, L.; Priest, D. G.; Ohno, M.

2024-12-07 biophysics
10.1101/2024.12.05.625331 bioRxiv
Show abstract

Light sheet microscopy combines high sensitivity and imaging speeds with low photodamage, however a remaining hurdle to its widespread adoption is its requirement for special sample preparation. Our new microscope - planar illumination microscope for single molecule imaging for all purpose (PISA) - significantly relieves these restrictions to enable light sheet imaging of coverslip-mounted samples as commonly analyzed in biological microscopes. We show how this technology now permits high-throughput imaging using multi-well plates. Further, the ability of 3D single molecule imaging in microlitre or mm3 scale volumes over the coverslip improves the sensitivity of biochemical gel-based fluorometric assays to atto/zepto molar levels. PISA will facilitate widespread adoption of light sheet imaging for a variety of biological, biochemical and medical investigations.

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