Compact, high-resolution fluorescence and brightfield microscope and methods of use

Abstract

The present invention provides a compact, inexpensive fluorescence microscope capable of high-resolution imaging with high light throughput suitable for use in both laboratory and field environments, and methods of use. A simple and inexpensive fluorescence microscope allows health care workers to perform various medical assays at the point of care instead of having to collect and transport biological samples to distant labs, and subsequently return the results to the patient. The microscope of the present invention is also useful for educational use and field use, and other uses as well.

1 . An inverted fluorescence microscope, comprising:
 a. at least one stage for placing at least one sample for observation;   b. a at least one epifluorescence illumination and detection system, comprising:
 i. at least one filter set below said stage, comprising:
 a) at least one excitation filter; 
 b) at least one dichroic mirror; and 
 c) at least one emission filter; 
 
 ii. at least one objective lens between said stage and said filter set; 
 iii. at least one image sensor below said filter set; 
 iv. at least one epi-illumination system below said stage, comprising:
 a) at least one source of epi-illumination light, 
 b) at least one condenser lens; and 
 c) at least one said excitation filter of said filter set, 
 
 v. at least one imaging barrel below said dichroic mirror in said filter set, comprising:
 a) said emission filter; 
 b) at least one projection lens; and 
 c) said image sensor; 
 
   c. at least one focusing assembly that attaches said epi-fluorescence illumination and detection system to said stage; and   d. at least one host computer comprising at least one program to control image acquisition of at least one image from said image sensor.   
     
     
         2 . The inverted fluorescence microscope of  claim 1 , wherein said stage comprises an x-y clamping caliper system for horizontal movement of said sample. 
     
     
         3 . The inverted fluorescence microscope of  claim 1 , wherein said epifluorescence illumination and detection system lacks a direct optical human interface. 
     
     
         4 . The inverted fluorescence microscope of  claim 1 , further comprising at least one sample cover to shield said stage from an outside light source or a stray light source. 
     
     
         5 . The inverted fluorescence microscope of  claim 4 , wherein said sample cover can be displaced to expose said stage. 
     
     
         6 . The inverted fluorescence microscope of  claim 1 , wherein said epi-illumination system extends in a generally horizontal direction from said filter set. 
     
     
         7 . The inverted fluorescence microscope of  claim 1 , wherein said excitation filter provides non-vergent or vergent focus of said source of epi-illumination light on said sample. 
     
     
         8 . The inverted fluorescence microscope of  claim 1 , wherein in said epifluorescence illumination and detection system, said source of epi-illumination light, said condenser lens, and said excitation filter are mounted in at least one illumination tube as a generally horizontal extension from at least one filter cube with at least one optical axis extending horizontally toward said filter set from said source of epi-illumination light, and light emitted from said source of epi-illumination light being directed to said stage by way of said dichroic mirror. 
     
     
         9 . The inverted fluorescence microscope of  claim 8 , wherein said filter cube comprises an about  45  degree slanted end of said illumination tube, and comprises said dichroic mirror centered at an optical axis extending from said objective lens to said projection lens and focuses at least one sample image on said image sensor, wherein along said optical axis said emission filter is provided between said dichroic mirror and said projection lens. 
     
     
         10 . The inverted fluorescence microscope of  claim 1 , wherein said imaging barrel focuses at least one filtered image onto said image sensor. 
     
     
         11 . The inverted fluorescence microscope of  claim 1 , wherein said imaging barrel is mounted in at least one tube extending in a generally vertical direction away from said filter set and focuses at least one filtered image of said sample onto said image sensor. 
     
     
         12 . The inverted fluorescence microscope of  claim 1 , wherein in said focusing assembly, the field of view does not substantially shift during focusing and that variation of focus due to mechanical vibration is minimized. 
     
     
         13 . The inverted fluorescence microscope of  claim 1 , wherein said source of epi-illumination light comprises at least one light-emitting diode (LED). 
     
     
         14 . The inverted fluorescence microscope of  claim 1 , wherein said source of epi-illumination light comprises at least one laser diode. 
     
     
         15 . The inverted fluorescence microscope of  claim 1 , wherein said program controls at least one image display from said image sensor. 
     
     
         16 . The inverted fluorescence microscope of  claim 1 , wherein said program configures at least one image display from said image sensor. 
     
     
         17 . The inverted fluorescence microscope of  claim 1 , wherein said image sensor detects multiple fluorescent emissions of differing wavelengths. 
     
     
         18 . The inverted fluorescence microscope of  claim 17 , wherein said multiple fluorescent emissions are from different fluorescent labels in said sample. 
     
     
         19 . The inverted fluorescence microscope of  claim 1 , wherein said excitation filter provides a plurality of wavelengths or range of wavelengths from said source of epi-illumination light. 
     
     
         20 . The inverted fluorescence microscope of  claim 1 , wherein said dichroic mirror provides excitation light towards said sample and transmits emission light towards said image sensor. 
     
     
         21 - 31 . (canceled)