Multi-Exposure Imaging for Automated Fluorescent Microscope Slide Scanning

1 . A method of forming an image of a fluorescent microscope slide, comprising:

acquiring an image of a fluorescent microscope slide at a first exposure level resulting in a first acquired image at a first contrast level;

acquiring at least one additional image of said fluorescent microscope slide each at an exposure level different from each other and different from the first exposure, resulting in different acquired images at different contrast levels;

forming a composite image of said fluorescent microscope slide by forming a weighted sum of the intensity values of corresponding pixels of each of said images, wherein said composite image has a composite contrast level over the entire composite image that represents a weighted average of said contrast levels.

2 . The method of claim 1 wherein the weight given to each of said pixels of each of said images is calculated in part from a localized contrast measure around that pixel.

3 . The method of claim 2 wherein a higher weight is given to a pixel whose localized contrast is higher.

4 . The method of claim 2 wherein the localized contrast measure is determined over a pre-defined sub-area of an image including said pixel.

5 . The method of claim 4 further comprising forming a range image for each image acquired at said exposure levels.

6 . The method of claim 5 wherein said range image is processed to determine said localized contrast measure.

7 . A method of forming an image of a fluorescent microscope slide, comprising:

using multi-exposure auto-focus at one or more points spaced at or around one or more sub-areas within said microscope slide;

generating a map of auto-focus values per sub-area of said microscope slide;

acquiring an image of a fluorescent microscope slide using said map of auto-focus values at a first exposure level resulting in a first acquired image at a first contrast level;

acquiring additional images of said fluorescent microscope slide using said map of auto-focus values, said images acquired at an exposure level different from each other and different from the first exposure, resulting in different acquired images having different contrast levels;

forming a composite image of said fluorescent microscope slide by forming a weighted sum of the intensity values of corresponding pixels of each of said images, wherein said composite image has a composite contrast level over the entire composite image that represents a weighted average of said contrast levels.

8 . The method of claim 7 wherein the weight given to each of said pixels of each of said images is calculated in part from a localized contrast measure around that pixel.

9 . The method of claim 8 wherein a higher weight is given to a pixel whose localized contrast is higher.

10 . The method of claim 8 wherein the localized contrast measure is determined over a pre-defined sub-area of an image including said pixel.

11 . The method of claim 10 further comprising forming a range image for each image acquired at said exposure levels.

12 . The method of claim 11 wherein said range image is processed to determine said localized contrast measure.

13 . A fluorescent image system comprising:

a source of illumination;

a microscope system;

a CCD camera for acquiring digitized images;

a computing unit for storing and processing said digitized images of a fluorescent microscope slide, said computing unit executing a method so as to cause:

a CCD camera image of said fluorescent microscope slide to be acquired at a first exposure level resulting in a first contrast level;

additional images of said fluorescent microscope slide to be acquired each at an exposure level different from each other and different from the first exposure, resulting in different contrast levels;

a composite image of said fluorescent microscope slide to be formed as a weighted sum of the intensity values of corresponding pixels of each of said images, wherein said composite image has a composite contrast level over the entire composite image that represents a weighted average of said contrast levels.

14 . The system as in claim 13 wherein said microscope system comprises an epi-fluorescence system comprising:

one or more excitation wavelength filters;

a dichroic mirror;

a system of lenses for directing the light beam;

one or more emission wavelength filters; and

a microscope objective.

15 . The system as in claim 13 wherein said microscope system comprise a trans-illuminating system comprising:

one or more excitation wavelength filters;

a mirror;

a system of lenses for directing the light beam;

one or more emission wavelength filters; and

a microscope objective.

16 . The system as in claim 13 wherein said computing unit executes a method so as to calculate the weight given to each of said pixels of each of said images in part from a localized contrast measure around that pixel.

17 . The system as in claim 16 wherein a higher weight is given to a pixel whose localized contrast is higher.

18 . The system as in claim 16 wherein the localized contrast measure is determined over a pre-defined sub-area of an image including said pixel.

19 . The system as in claim 18 wherein said computing unit executes a method so as to form a range image for each image acquired at said exposure levels.

20 . The system as in claim 19 wherein said computing unit executes a method so as to process said range image to determine said localized contrast measure.

21 . The system as in claim 13 wherein said computing unit is configured to use a map of auto-focus values to adjust said CCD camera focus per said slide area; said map created by multi-exposure auto-focus at one or more points spaced at or around one or more sub-areas within said microscope slide.