System for creating microscopic digital montage images

Abstract

An imaging apparatus. The imaging apparatus may find an area in which a specimen is present, then focus on the specimen and capture images of the specimen during continuous stage motion.

1 . A focusing method, comprising: 
 focusing a microscope optic on a plurality of points of a specimen to determine a focal length for each point;    fitting a surface to the points at the focal lengths; and    capturing a plurality of microscopic images of portions of the specimen while focusing on each microscopic image based on the fitted surface.    
     
     
         2 . The focusing method of  claim 1 , wherein the fitted surface is curved.  
     
     
         3 . The focusing method of  claim 1 , wherein the fitted surface is planar.  
     
     
         4 . The focusing method of  claim 1 , wherein the fitted surface is described by a model.  
     
     
         5 . The focusing method of  claim 1 , wherein the fitted surface is described by an equation.  
     
     
         6 . The focusing method of  claim 1 , wherein the fitted surface is a focal surface.  
     
     
         7 . The focusing method of  claim 1 , wherein the fitted surface includes at least comprises an area equivalent to an area of the specimen.  
     
     
         8 . The focusing method of  claim 1 , wherein the captured microscopic images are adjacent.  
     
     
         9 . The focusing method of  claim 1 , further comprising capturing a first image of the specimen; and 
 selecting the plurality of points on which to focus the microscopic optic from the first image.    
     
     
         10 . The focusing method of  claim 9 , wherein the first image is a macroscopic image.  
     
     
         11 . The focusing method of  claim 9 , wherein the first image is captured at a lower magnification than that of any of the microscopic images.  
     
     
         12 . The focusing method of  claim 9 , wherein the plurality of points are selected based on a relative intensity of the image at each of the points.  
     
     
         13 . The focusing method of  claim 1 , wherein the plurality of points are selected based on their distribution across the specimen.  
     
     
         14 . The focusing method of  claim 1 , wherein the specimen is divided into regions and the plurality of points are each selected from a different one of the regions.  
     
     
         15 . The focusing method of  claim 1 , further comprising calculating an error function to determine a fit accuracy of the fitted surface.  
     
     
         16 . The focusing method of  claim 1 , further comprising: 
 selecting an additional point;    determining another focal length for the additional point; and    fitting another surface to the plurality of points at the focal lengths and the additional point at the other focal length.    
     
     
         17 . The focusing method of  claim 1 , wherein the specimen is moved continuously while a plurality of adjacent images of the specimen are captured; and 
 wherein the microscope optic is focused based on the focal length of the fitted surface for each of the adjacent images to be captured while the specimen is moved.    
     
     
         18 . An imaging apparatus, comprising: 
 a motorized stage;    a camera focused relative to the motorized stage; and    a processor coupled to the camera, wherein the processor contains instructions which, when executed by the processor, cause the processor to: 
 capture a low resolution image that is incident on the camera, wherein the low resolution image includes a plurality of pixels, the pixels having a characteristic;  
 establish the characteristic for each pixel;  
 determine which of the pixels contain a target image based on the characteristic of the pixels and establish a target area that includes those pixels;  
 transpose the position of the target area into a plurality of stage coordinates; and  
 capture a high resolution image that is incident on the camera at each of the stage coordinates.  
   
     
     
         19 . The imaging apparatus of  claim 18 , wherein the characteristic includes pixel intensity.  
     
     
         20 . The imaging apparatus of  claim 18 , wherein the characteristic includes pixel color.  
     
     
         21 . The imaging apparatus of  claim 18 , wherein the processor determines which pixels contain the target image based on a relative intensity of the pixels and further: 
 determines a mean intensity of the pixels;    compares the intensity of each of the pixels to the mean intensity; and    divides the pixels into a group of non-target image pixels having high intensities and a group of target image pixels having intensities lower than the high intensities.    
     
     
         22 . The imaging apparatus of  claim 18 , wherein the processor determines which of the pixels contain the target image based on a relative intensity of the pixels and further: 
 determines, for the pixels, an intensity standard deviation that provides an amount of variation in pixel intensity;    compares the intensity of each pixel to the intensity standard deviation; and    divides the pixels into a group of non-target image pixels having low standard deviations and a group of target image border pixels having standard deviations that are greater than any of the low standard deviations.    
     
     
         23 . The imaging apparatus of  claim 18 , further comprising a pulsed light directed toward the motorized stage.  
     
     
         24 . The imaging apparatus of  claim 18 , further comprising a stage position sensor adjacent the motorized stage.  
     
     
         25 . An imaging apparatus, comprising: 
 a motorized stage;    a camera having a lens directed toward the motorized stage; and    a processor coupled to the camera, wherein the processor contains instructions which, when executed by the processor, cause the processor to: 
 select at least three points of a sample adjacent the motorized stage;  
 determine a stage position for each of the selected points;  
 focus the camera on each of the selected points;  
 determine an object distance from the camera lens to the sample at each of the selected points; and  
 develop a focus surface based on the stage position and the object distance for the selected points.  
   
     
     
         26 . The imaging apparatus of  claim 25 , wherein the selecting at least three points of a sample adjacent the motorized stage includes selecting points dependent on a characteristic of an image of the sample at those points.  
     
     
         27 . The imaging apparatus of  claim 25 , wherein the selecting points dependent on a characteristic of the image of the sample at those points includes selecting the darkest regions.  
     
     
         28 . The imaging apparatus of  claim 25 , wherein the selecting points dependent on a characteristic of the image of those regions includes selecting the lightest regions.  
     
     
         29 . The imaging apparatus of  claim 25 , wherein the selecting points dependent on a characteristic of the image of those regions includes selecting points having a high contrast relative to the regions.  
     
     
         30 . The imaging apparatus of  claim 25 , wherein when the processor selects at least three points of a sample adjacent the motorized stage, the processor further: 
 determines a distribution of the at least three selected points within the sample;    determines whether at least one of the selected points lies within each of at least two predetermined areas; and    selects additional points until at least one of the additional points lies within each predetermined area.    
     
     
         31 . An imaging apparatus, comprising: 
 a motorized stage;    a camera focused relative to the motorized stage;    a stage position sensor adjacent the motorized stage; and    a pulsed light directed toward the motorized stage and coupled to the stage position sensor such that the pulsed light illuminates in response to the stage position sensor.    
     
     
         32 . The imaging apparatus of  claim 31 , further comprising a processor coupled to the camera, the pulsed light, and the stage position sensor, wherein the processor contains instructions which, when executed, cause the processor to: 
 initiate motion of the motorized stage;    energize the pulsed light when the stage position sensor indicates the motorized stage is in a predetermined position; and    capture an image by way of the camera while the pulsed light is energized.    
     
     
         33 . The imaging apparatus of  claim 32 , wherein the motorized stage moves continuously while the images are captured.