Abstract: The present invention relates to systems and methods for sequential operation of staining, imaging and sectioning of tissue samples by a processing system. After each layer of the sample is removed by the sectioning system, the system automatically stains the exposed surface of a sample to a depth to enable imaging of the remaining tissue. The system then repeats the sectioning, staining and imaging steps in sequence to image the sample.

Abstract: In accordance with preferred embodiments of the present invention, a method for imaging tissue, for example, includes the steps of mounting the tissue on a computer controlled stage of a microscope, determining volumetric imaging parameters, directing at least two photons into a region of interest, scanning the region of interest across a portion of the tissue, imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest, sectioning the portion of the tissue, capturing the sectioned tissue, and imaging a second plurality of layers of the tissue in a second plurality of volumes of the tissue in the region of interest, and capturing each sectioned tissue, detecting a fluorescence image of the tissue due to said excitation light; and processing three-dimensional data that is collected to create a three-dimensional image of the region of interest.

Abstract: The present invention relates to systems and methods for sequential operation of staining, imaging and sectioning of tissue samples by a processing system. After each layer of the sample is removed by the sectioning system, the system automatically stains the exposed surface of a sample to a depth to enable imaging of the remaining tissue. The system then repeats the sectioning, staining and imaging steps in sequence to image the sample.

Abstract: A microtome method and apparatus includes a microtome blade configured to oscillate in a direction transverse to a direction of advancing a cut, and a first flexure to support and guide the blade. The first flexure is compliant in the transverse direction while being stiff in the cut direction. A second flexure operatively engaged at one end portion with the first flexure, is stiff in the transverse direction while being compliant in the cut direction. The other end portion of the second flexure is rotatably engaged by an eccentric driven by a rotatable actuator, which oscillates the blade in the transverse direction while effectively isolating non-transverse motion from the blade. The second flexure is configured to move independently of any guides or other stationary objects during oscillation.

Abstract: A microtome method and apparatus includes a microtome blade configured to oscillate in a direction transverse to a direction of advancing a cut, and a first flexure to support and guide the blade. The first flexure is compliant in the transverse direction while being stiff in the cut direction. A second flexure operatively engaged at one end portion with the first flexure, is stiff in the transverse direction while being compliant in the cut direction. The other end portion of the second flexure is rotatably engaged by an eccentric driven by a rotatable actuator, which oscillates the blade in the transverse direction while effectively isolating non-transverse motion from the blade. The second flexure is configured to move independently of any guides or other stationary objects during oscillation.

Abstract: A microtome method and apparatus includes a microtome blade configured to oscillate in a direction transverse to a direction of advancing a cut, and a first flexure to support and guide the blade. The first flexure is compliant in the transverse direction while being stiff in the cut direction. A second flexure operatively engaged at one end portion with the first flexure, is stiff in the transverse direction while being compliant in the cut direction. The other end portion of the second flexure is rotatably engaged by an eccentric driven by a rotatable actuator, which oscillates the blade in the transverse direction while effectively isolating non-transverse motion from the blade. The second flexure is configured to move independently of any guides or other stationary objects during oscillation.

Abstract: A microtome method and apparatus includes a microtome blade configured to oscillate in a direction transverse to a direction of advancing a cut, and a first flexure to support and guide the blade. The first flexure is compliant in the transverse direction while being stiff in the cut direction. A second flexure operatively engaged at one end portion with the first flexure, is stiff in the transverse direction while being compliant in the cut direction. The other end portion of the second flexure is rotatably engaged by an eccentric driven by a rotatable actuator, which oscillates the blade in the transverse direction while effectively isolating non-transverse motion from the blade. The second flexure is configured to move independently of any guides or other stationary objects during oscillation.

Abstract: A microtome method and apparatus includes a microtome blade configured to oscillate in a direction transverse to a direction of advancing a cut, and a first flexure to support and guide the blade. The first flexure is compliant in the transverse direction while being stiff in the cut direction. A second flexure operatively engaged at one end portion with the first flexure, is stiff in the transverse direction while being compliant in the cut direction. The other end portion of the second flexure is rotatably engaged by an eccentric driven by a rotatable actuator, which oscillates the blade in the transverse direction while effectively isolating non-transverse motion from the blade. The second flexure is configured to move independently of any guides or other stationary objects during oscillation.

Abstract: In accordance with preferred embodiments of the present invention, a method for imaging tissue, for example, includes the steps of mounting the tissue on a computer controlled stage of a microscope, determining volumetric imaging parameters, directing at least two photons into a region of interest, scanning the region of interest across a portion of the tissue, imaging layers of the tissue, sectioning a portion of the tissue, capturing the sectioned tissue, and imaging additional layers of the tissue in a second volume of the tissue, and capturing each portion of sectioned tissue, and processing three-dimensional data that is collected to create a three-dimensional image of the region of interest. Further, captured tissue sections can be processed, re-imaged, and indexed to their original location in the three dimensional image.