Structured illumination in inverted light sheet microscopy

Patent metadata

Field	Value
Publication number	US-10401605-B2
Application number	US-201715843692-A
Country	US
Kind code	B2
Filing date	Dec 15, 2017
Priority date	May 2, 2017
Publication date	Sep 3, 2019
Grant date	Sep 3, 2019

Abstract

A system, including a structured illumination stage to provide a spatially modulated imaging field is provided. The system further includes a spatial frequency modulation stage to adjust the frequency of the spatially modulated imaging field, a sample interface stage to direct the spatially modulated imaging field to a sample, and a sensor configured to receive a plurality of fluorescence emission signals from the sample. The system also includes a processor configured to reduce a sample scattering signal and to provide a fluorescence emission signal from a portion of the sample including the spatially modulated imaging field. A method for using the above system to form an image of the sample is also provided.

First claim

The invention claimed is:

1. A system, comprising:
a structured illumination stage to provide a spatially modulated imaging field;
a spatial frequency modulation stage to adjust a frequency of the spatially modulated imaging field;
a sample interface stage to direct the spatially modulated imaging field to a sample;
a sensor configured to receive a plurality of fluorescence emission signals from the sample; and
a processor configured to reduce a sample scattering signal and to provide a fluorescence emission signal from a portion of the sample including the spatially modulated imaging field.

2. The system of claim 1 , wherein the structured illumination stage comprises an anamorphic optical component to form the spatially modulated imaging field in a sheet of light.

3. The system of claim 1 , further comprising a phase shifting stage to adjust the phase of the spatially modulated imaging field, the structured illumination stage comprising a spatial light modulator configured to spatially separate two diffracted beams having a selected phase shift between them, to form the spatially modulated imaging field.

4. The system of claim 1 , wherein the spatial frequency modulation stage comprises a movable optical element configured to adjust an angle of incidence formed between a first diffracted illumination beam and a second diffracted illumination beam provided by the structured illumination stage, thereby to adjust a spatial frequency of an interference pattern between the first diffracted illumination beam and the second diffracted illumination beam.

5. A method for forming an image of a sample, comprising:
providing a spatially modulated imaging field;
adjusting a frequency of the spatially modulated imaging field;
directing, with a sample interface, the spatially modulated imaging field to a sample;
receiving a plurality of fluorescence emission signals from the sample; and
combining the plurality of fluorescence emission signals to reduce a sample scattering signal and to obtain an image of the sample.

6. The method of claim 5 , wherein receiving a plurality of fluorescence emission signals from the sample comprises receiving a first fluorescence emission signal from a structured illumination field having a spatial frequency and a first phase, receiving a second fluorescence emission signal having the spatial frequency with a second phase, and receiving a third fluorescence emission signal having the spatial frequency with a third phase.

7. The method of claim 5 , wherein receiving a plurality of fluorescence emission signals comprises receiving the plurality of fluorescence emission signals along a line intersecting a plane including the spatially modulated imaging field and a plane of motion of the sample.

8. The method of claim 5 , further comprising moving the sample along a plane oblique to a plane including the spatially modulated imaging field, and obtaining a plurality of images of the sample to form a three-dimensional image of the sample.

9. The method of claim 5 , wherein adjusting a frequency of the spatially modulated imaging field comprises linearly displacing an optical element to interfere two optical beams at a selected angle.

10. The method of claim 5 , wherein receiving a plurality of fluorescence emission signals from the sample comprises receiving a first fluorescence signal with the spatially modulated imaging field having a first phase, receiving a second fluorescence signal with the spatially modulated imaging field having a second phase, and receiving a third fluorescence emission signal with the spatially modulated imaging field having a third phase.

Assignees

• Screen Holdings Co Ltd
• Univ Leland Stanford Junior

Inventors

• Landry Joseph Russell
• Itoh Ryosuke
• Mandella Michael J
• Solgaard Olav

Classifications

• G02B21/367

  providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison · CPC title

• G01N2201/0675

  SLM · CPC title

• G02B21/0032

  Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers (G02B21/0036 - G02B21/008; means for illumination of specimens in general G02B21/06) · CPC title

• G02B21/0076

  arrangements using fluorescence or luminescence · CPC title

• G01N21/6458

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title