Fluorescence biopsy specimen imager and methods
An imaging device is described that uses multiple cameras to image a biological sample on a turntable bathed in white light or fluorescing due to a systemically administered dye. Fluorescence farther away from an excitation light source can be compensated upon determining a 3-D position of portions of the sample. The turntable is turned and tilted in order to take enough images to prepare an animation of the sample. In a graphical user interface, the animation can be stopped, zoomed, and tilted per a user's gesture, touch, tablet-tilting, or other commands. The image manipulation can be with touch gestures entered using a sterilizable or disposable touch pen.
1. An apparatus for imaging a biological sample with fluorescence, the apparatus comprising:
a rotatable imaging stage adapted for supporting at least a portion of a biological sample within an imaging volume;
a first camera;
a monochromatic second camera; and
a switchable mounting system configured to alternatingly position the first camera and the second camera to receive light reflecting or emitting from the biological sample.
2. The apparatus of claim 1 further comprising:
a fluorescence excitation light source configured to illuminate the imaging volume.
3. The apparatus of claim 1 further comprising:
a preview third camera having an optical system focused on the imaging volume.
4. The apparatus of claim 1 further comprising:
a transparent window having a plurality of equally spaced marks at predetermined intervals, the transparent window attached to the imaging stage.
5. The apparatus of claim 4 further comprising:
a computer processor operatively connected with a machine-readable non-transitory medium embodying information indicative of instructions for causing the computer processor to perform operations comprising:
taking reflected light images of a biological sample on the stage using the first camera, the reflected light images including the equally spaced marks;
collecting fluorescence images of the biological sample using the second camera;
detecting the equally spaced marks in the reflected light images or the fluorescence images; and
extrapolating the equally spaced marks in order to determine a three-dimensional position of a portion of the biological sample.
6. An apparatus for imaging a biological sample with fluorescence, the apparatus comprising:
a rotatable imaging stage adapted for supporting at least a portion of a biological sample within an imaging volume;
a transparent window having a plurality of equally spaced marks at predetermined intervals, the transparent window attached to the imaging stage;
a first camera;
a monochromatic second camera; and
a partially reflective mirror, the mirror configured to reflect a portion of an image of the at least a portion of the biological sample to one of the cameras, and the mirror configured to transmit a portion of the image to the other of the cameras.
7. The apparatus of claim 6 further comprising:
a fluorescence excitation light source configured to illuminate the imaging volume.
8. The apparatus of claim 6 further comprising:
a preview third camera having an optical system focused on the imaging volume.
9. The apparatus of claim 6 further comprising:
a computer processor operatively connected with a machine-readable non-transitory medium embodying information indicative of instructions for causing the computer processor to perform operations comprising:
taking reflected light images of a biological sample on the stage using the first camera, the reflected light images including the equally spaced marks;
collecting fluorescence images of the biological sample using the second camera;
detecting the equally spaced marks in the reflected light images or the fluorescence images; and
extrapolating the equally spaced marks in order to determine a three-dimensional position of a portion of the biological sample.
10. An apparatus for imaging a biological sample with fluorescence, the apparatus comprising:
a rotatable imaging stage adapted for supporting at least a portion of a biological sample within an imaging volume;
a first camera;
a monochromatic second camera;
a flip mirror; and
an optical system configured to present an image of the at least a portion of the biological sample to the flip mirror, the flip mirror having a first configuration for reflecting the image to the first camera and a second configuration for reflecting the image to the second camera.
11. The apparatus of claim 10 further comprising:
a fluorescence excitation light source configured to illuminate the imaging volume.
12. The apparatus of claim 10 further comprising:
a preview third camera having an optical system focused on the imaging volume.
13. The apparatus of claim 10 further comprising:
a transparent window having a plurality of equally spaced marks at predetermined intervals, the transparent window attached to the imaging stage.
14. The apparatus of claim 13 further comprising:
a computer processor operatively connected with a machine-readable non-transitory medium embodying information indicative of instructions for causing the computer processor to perform operations comprising:
taking reflected light images of a biological sample on the stage using the first camera, the reflected light images including the equally spaced marks;
collecting fluorescence images of the biological sample using the second camera;
detecting the equally spaced marks in the reflected light images or the fluorescence images; and
extrapolating the equally spaced marks in order to determine a three-dimensional position of a portion of the biological sample.
15. A method of correcting an image for divergence of fluorescence excitation light, the method comprising:
illuminating a biological sample on a stage with a fluorescence excitation light source;
taking a first fluorescence image of the biological sample using a camera;
rotating the stage from a first angle to a second angle;
collecting a second fluorescence image of the biological sample using the camera;
determining from the fluorescence images a three-dimensional position of a portion of the biological sample;
ascertaining a divergence of the fluorescence excitation light;
acquiring a pixel reading of one of the images; and
applying a correction to the pixel reading based on the three-dimensional position and the divergence.
16. The method of claim 15 wherein the correction is based on a predetermined, calculated divergence of the fluorescence excitation light.