Compressed acquisition of microscopic images
A microscope for computational imaging may include an illumination source configured to illuminate a sample with a plurality of wavelengths, an image sensor, an objective lens to image the sample onto the image sensor, and a processor operatively coupled to the illumination assembly and the image sensor. The processor may be configured to acquire a first image dataset from the sample illuminated using a first set of illumination conditions at a first wavelength. The processor may also be configured to acquire a second image dataset from the sample illuminated using a second set of illumination conditions having a second number of illumination conditions at a second wavelength. The second set of illumination conditions comprises fewer illumination conditions than the first set in order to decrease acquisition time. The processor may be configured to combine the first and second image datasets into a computationally reconstructed image of the sample.
1 . A method for generating a computationally reconstructed image, comprising:
acquiring, with an image sensor, a first image dataset from a sample illuminated using a first set of illumination conditions comprising a first number of illumination conditions each comprising a first wavelength, the first image dataset comprising a plurality of images comprising a first spatial frequency bandwidth;
acquiring, with the image sensor, a second image dataset from the sample illuminated using a second set of illumination conditions comprising a second number of illumination conditions, each comprising a second wavelength, the second image dataset comprising one or more images comprising a second spatial frequency bandwidth, wherein the first number of illumination conditions is larger than the second number of illumination conditions; and
combining the first image dataset and the second image dataset into a computationally reconstructed image of the sample, wherein the computationally reconstructed image comprises a spatial frequency bandwidth greater than the first spatial frequency bandwidth of the first image dataset and the second spatial frequency bandwidth of the second image dataset.
2 . The method of claim 1 , wherein the first spatial frequency bandwidth corresponds to the first wavelength and the second spatial frequency bandwidth corresponds to the second wavelength, and wherein the first spatial frequency bandwidth is greater than the second spatial frequency bandwidth.
3 . The method of claim 2 , wherein the computationally reconstructed image comprises one or more of an increased contrast or an aberration correction for the second wavelength based at least in part on the first image dataset from the first wavelength.
4 . The method of claim 1 , further comprising:
providing the reconstructed image on a display with the first spatial frequency bandwidth and a first user perceivable color corresponding to the first wavelength and the second spatial frequency bandwidth and a second user perceivable color corresponding to the second wavelength, the first spatial frequency bandwidth greater than the second spatial frequency bandwidth and a spatial frequency bandwidth of the plurality of images acquired by the image sensor with the first set of illumination conditions at the first wavelength.
5 . The method of claim 1 , wherein the first number is larger than the second number by at least a factor of two and a second acquisition time associated with the second image dataset is no more than half of a first acquisition time associated with the first image dataset and wherein the spatial frequency bandwidth of the computationally reconstructed image is greater than a spatial frequency bandwidth of each of a plurality of images acquired with the first set of illumination conditions by at least a factor of 1.5.
6 . The method of claim 1 , wherein the computationally reconstructed image comprises one or more of a correction for optical aberrations, or an increase in image contrast.
7 . The method of claim 6 , wherein the image sensor comprise a spatial frequency bandwidth and the spatial frequency bandwidth of the computationally reconstructed image is greater than the spatial frequency bandwidth of the image sensor divided by a magnification of an image of the sample onto the image sensor.
8 . The method of claim 6 , wherein each of the first plurality of images comprises the first spatial frequency bandwidth, and the spatial frequency bandwidth of the computationally reconstructed image is greater than the first spatial frequency bandwidth of said each of the first plurality of images.
9 . The method of claim 6 , wherein the correction of optical aberrations is provided by separating aberration information from sample information, so as to decrease an effect of optical aberrations on the computationally reconstructed image.
10 . The method of claim 6 , wherein the increased image contrast of the computationally reconstructed image is provided by computationally amplifying high spatial frequencies of the reconstructed image to better represent the sample.
11 . The method of claim 1 , wherein the plurality of images and the one or more images are processed together to generate the computationally reconstructed image.
12 . The method of claim 1 , wherein the computationally reconstructed image comprises a color image, the color image comprising two or more of a red channel, a green channel, or a blue channel.
13 . The method of claim 12 , wherein the first wavelength corresponds to one of the red channel, the green channel or the blue channel and the second wavelength corresponds to another one of the red channel, the blue channel or the green channel.
14 . The method of claim 13 , wherein the computationally reconstructed image is shown on a display with the first spatial frequency bandwidth for a first color channel corresponding to the first wavelength and with the second spatial frequency bandwidth for a second color channel corresponding to the second wavelength, the first spatial frequency bandwidth greater than the second spatial frequency bandwidth.
15 . The method of claim 14 , wherein the first wavelength comprises green light, the first channel comprises the green channel, the second wavelength comprises red light or blue light and the second color channel comprises the red channel or the blue channel and wherein the green channel is shown on the display with the first spatial frequency bandwidth and the red channel or the blue channel is shown on the display with the second spatial frequency bandwidth.
16 . The method of claim 15 , wherein the computationally reconstructed image comprises the red channel, the green channel and the blue channel and wherein the second wavelength comprises red light corresponding to the red channel and a third wavelength comprises blue light corresponding to a third channel wherein the third channel is shown on the display with a third spatial frequency bandwidth less than the first spatial frequency bandwidth.
17 . The method of claim 1 , further comprising determining the first wavelength.
18 . The method of claim 17 , wherein the first wavelength is user-defined.
19 . The method of claim 17 , wherein determining the first wavelength further comprises:
acquiring, with the image sensor, an initial image dataset of the sample illuminated with a plurality of wavelengths;
determining that a first image of the initial image dataset includes more information than a second image of the initial image dataset;
selecting, as the first wavelength, a first wavelength of the plurality of wavelengths corresponding to the first image; and
selecting, as the second wavelength, a second wavelength of the plurality of wavelengths corresponding to the second image.
20 . A microscope for image reconstruction, comprising:
an illumination assembly configured to illuminate a sample with a plurality of wavelengths at a plurality of angles;
an image sensor;
an objective lens to image the sample illuminated with the illumination assembly onto the image sensor; and
a processor operatively coupled to the illumination assembly and the image sensor, the processor configured with instructions to perform the method of claim 1 .
21 . The method of claim 1 , wherein the first number of illumination conditions comprises a plurality of angles.