High-throughput optical sectioning via line-scanning imaging with digital structured modulation

Abstract

Optical sectioning with high-throughput, a high signal-to-noise ratio (SNR), and submicrometer resolution is crucial, but challenging, to three-dimensional visualization of large biological tissue samples. Here we propose line-scanning imaging with digital structured modulation for optical sectioning. Our method generates images with a significantly improved SNR, compared to wide-field structured illumination microscopy (WF-SIM), without residual modulation artifacts. We image a <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" display="inline"> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mn>14.5</mml:mn> </mml:mrow> <mml:mspace width="thickmathspace"/> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mi mathvariant="normal">m</mml:mi> <mml:mi mathvariant="normal">m</mml:mi> </mml:mrow> <mml:mo>×</mml:mo> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mn>11.5</mml:mn> </mml:mrow> <mml:mspace width="thickmathspace"/> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mi mathvariant="normal">m</mml:mi> <mml:mi mathvariant="normal">m</mml:mi> </mml:mrow> </mml:math> horizontal view of mouse brain tissue at a pixel resolution of <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" display="inline"> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mn>0.32</mml:mn> </mml:mrow> <mml:mspace width="thickmathspace"/> <mml:mtext>µ</mml:mtext> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mi mathvariant="normal">m</mml:mi> </mml:mrow> <mml:mo>×</mml:mo> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mn>0.32</mml:mn> </mml:mrow> <mml:mspace width="thickmathspace"/> <mml:mtext>µ</mml:mtext> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mi mathvariant="normal">m</mml:mi> </mml:mrow> </mml:math> in 101 s, which, compared to WF-SIM, represents a significant improvement on imaging throughput. These results provide development opportunities for high-throughput, high-resolution large-area optical imaging methods.

Topics & Concepts