Systems and methods for illumination phase control in fluorescence microscopy

A phase control and microscopy technology, applied in the field of fluorescence microscopy, can solve the problems of low optical distortion and slow switching time of deformable windows (about tens to hundreds of milliseconds, etc.)

Active Publication Date: 2013-12-25
莱卡微系统 CMS +1
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Problems solved by technology

However, deformable windows transition times are rather slow (on the order of tens to hundreds of milliseconds) and extremely low optical distortion is difficult to achieve because the window is undergoing physical deformation

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  • Systems and methods for illumination phase control in fluorescence microscopy
  • Systems and methods for illumination phase control in fluorescence microscopy
  • Systems and methods for illumination phase control in fluorescence microscopy

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[0019] Various illumination phase control (“IPC”) are described along with a general description of three-dimensional structured illumination microscopy (“3D-SIM”). Compared with conventional wide-field fluorescence microscopes used in cell biology, 3D-SIM achieves a 1 / 2 increase in lateral and axial resolution. Unlike some competing super-resolution techniques, 3D-SIM does not require specialized fluorescent dyes or proteins. Biologists use 3D-SIM to achieve high resolution but retain the convenience and familiarity of fluorescent labeling. Illumination phase control provides the ability to shift and rotate the illumination pattern to capture multiple images of the subject. Higher resolution can be achieved by solving a system of equations to recover fine spatial details that are usually blurred by diffraction.

[0020] figure 1 A schematic representation of an example 3D-SIM instrument 100 is shown. There are many different types of SIM instruments and corresponding opti...

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Abstract

Illumination phase controls that provide precise and fast phase control of structured illumination patterns used in structure illumination microscopy are described. A coherent light source is used to generate a beam of coherent light that is split into at least three coherent beams of light. In one aspect, an illumination phase control is composed of at least one pair of rotatable windows to apply at least one phase shift to at least one of the beams. An objective lens is to receive the beams and focus the at least three beams to form an interference pattern. The phase control can be used to change the position of the interference pattern by changing the at least one phase shift applied to the at least one beam.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of Provisional Application No. 61 / 447,707, filed March 1, 2011. technical field [0003] The present disclosure relates to fluorescence microscopy, and in particular to systems for controlling the phase of excitation light illuminating a sample. Background technique [0004] Precise phase control of light sources in microscopy instruments is important for various optical techniques involving interference of separate beams, as well as polarization optics. For example, phase control is used in phase contrast microscopy, differential interference microscopy, and polarized light microscopy. In particular, precise and fast phase control is used in constructed illumination microscopes where phase control of interfering beams exploits precise changes in the path lengths of the separated beam paths on the order of fractional wavelengths (which for visible light corresponds to 10 nanometers ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G02B26/06G02B26/08
CPCG02B21/0032G02B21/0076G02B21/06G02B21/14G02B21/367G02B26/0875G02B27/106G02B27/1093G02B27/144G02B27/58
Inventor J.R.库珀
Owner 莱卡微系统 CMS
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