Microscope-based system and method for image-guided microscopic illumination
A system and method for image-guided microscopic illumination are provided. A processing module controls an imaging assembly such that a camera acquires an image or images of a sample in multiple fields of view, and the image or images are automatically transmitted to a processing module and processed by the first processing module automatically in real-time based on a predefined criterion so as to determine coordinate information of an interested region in each field of view. The processing module also controls an illuminating assembly to illuminate the interested region of the sample according to the received coordinate information regarding to the interested region, with the illumination patterns changing among the fields of view.
1 . A system for image-guided microscopic illumination of a biological sample comprising:
i) a microscope comprising a stage;
ii) an imaging assembly;
iii) an illuminating assembly;
iv) a first processing module operatively coupled to the microscope and configured to control the stage; and
v) a second processing module comprising a memory unit, wherein the second processing module is operatively coupled to the imaging assembly, the illuminating assembly, and the first processing module;
wherein the second processing module is configured to:
(a) control the imaging assembly to acquire at least one image of a first field of view of the biological sample;
(b) process the at least one image automatically based on a predefined criterion to determine coordinate information of one or more regions of interest in the first field of view; and
(c) control the illuminating assembly to illuminate the biological sample with a light pattern corresponding to the one or more regions of interest in the first field of view according to the coordinate information of the one or more regions of interest in the first field of view;
wherein after the one or more regions of interest in the first field of view is illuminated, the first processing module is configured to control the stage of the microscope to move to a subsequent field of view.
2 . The system of claim 1 , wherein the imaging assembly comprises a camera.
3 . The system of claim 1 , wherein the imaging assembly comprises a camera, an imaging light source, and a first shutter to control the imaging light source.
4 . The system of claim 1 , wherein the memory unit comprises a random access memory (RAM), a flash read-only memory (ROM), or a hard drive.
5 . The system of claim 4 , wherein the RAM comprises a dynamic random access memory (DRAM), a static random access memory (SRAM), or a zero-capacitor random access memory (Z-RAM).
6 . The system of claim 1 , wherein the second processing module is operatively coupled to a signal converter, wherein the signal converter is configured to convert the coordinate information to an analog signal.
7 . The system of claim 1 , wherein the illuminating assembly comprises an illuminating light source and a second shutter to control the illuminating light source.
8 . The system of claim 3 , wherein the first shutter is a non-mechanical shutter.
9 . The system of claim 7 , wherein the second shutter is a non-mechanical shutter.
10 . The system of claim 9 , wherein the non-mechanical shutter comprises an acousto-optic modulator (AOM) or an electro-optic modulator (EOM).
11 . A method for image-guided microscopic illumination of a biological sample comprising:
(a) moving a stage of a microscope to a first position using a first processing module to position the biological sample for imaging of a first field of view;
(b) controlling a camera of an imaging assembly with a second processing module to acquire an image of the first field of view of the biological sample;
(c) performing image processing of the image automatically in real-time based on a predefined criterion with the second processing module to determine one or more regions of interest in the image and obtain coordinate information of the one or more regions of interest;
(d) controlling an illuminating assembly to illuminate the first field of view of the biological sample with a light pattern corresponding to the one or more regions of interest according to the coordinate information of the one or more regions of interest with the second processing module; and
(e) after the one or more regions of interest in the first field of view is illuminated, controlling the stage of the microscope with the first processing module to move to a subsequent field of view.
12 . The method of claim 11 , further comprising, after (b) and prior to (c), transmitting the image of the first field of view of the biological sample automatically from the imaging assembly to the second processing module.
13 . The method of claim 11 , further comprising, after (c) and prior to (d), transmitting the coordinate information corresponding to the one or more regions of interest from the second processing module to the illuminating assembly.
14 . The method of claim 13 , wherein transmitting the coordinate information corresponding to the one or more regions of interest from the second processing module to the illuminating assembly comprises converting the coordinate information to an analog signal.