Methods and Compositions for Preparing Biological Specimens for Microscopic Analysis

Abstract

Methods and compositions are provided for preparing a biological specimen for microscopic analysis. These methods find many uses, for example in medicine and research, e.g., to diagnose or monitor disease or graft transplantation, to study healthy or diseased tissue, to screen candidate agents for toxicity and efficacy in disease modification. Also provided are reagents, devices, kits and systems thereof that find use in practicing the subject methods.

1 .- 53 . (canceled) 
     
     
         54 . A system for preparing a biological specimen for imaging, the system comprising an electrophoretic tissue clearing device for electrophoresing a three-dimensional hydrogel-embedded biological specimen to substantially remove a plurality of cellular components from the specimen before imaging of the hydrogel-embedded biological specimen, wherein the device comprises:
 an electrophoresis chamber for containing the hydrogel-embedded biological specimen;   a plurality of electrodes;   a power supply; and   a temperature-controlled buffer circulator.   
     
     
         55 . The system according to  claim 54 , wherein the device further comprises a buffer filtering component. 
     
     
         56 . The system according to  claim 54 , wherein the device further comprises a plurality of fluid inlet and/or outlet ports. 
     
     
         57 . The system according to  claim 54 , wherein the device further comprises a component configured to support the hydrogel-embedded biological specimen. 
     
     
         58 . The system according to  claim 57 , wherein the component is configured to support the hydrogel-embedded biological specimen in a position that is substantially inside an electric field generated between two or more of the electrodes. 
     
     
         59 . The system according to  claim 54 , wherein one or more of the electrodes comprises an expansion component for increasing the size of an electric field generated by the electrodes. 
     
     
         60 . The system according to  claim 59 , wherein the expansion component comprises one or more S-shaped bends. 
     
     
         61 . The system according to  claim 54 , wherein the length and the width of the one or more electrodes are approximately equal. 
     
     
         62 . The system according to  claim 54 , further comprising a lid that forms a fluid-tight and/or air-tight seal with the electrophoresis chamber. 
     
     
         63 . The system according to  claim 54 , further comprising a buffer solution that comprises an ionic surfactant. 
     
     
         64 . The system according to  claim 63 , wherein the ionic surfactant is sodium dodecyl sulfate. 
     
     
         65 . The system according to  claim 54 , further comprising the hydrogel-embedded biological specimen. 
     
     
         66 . The system according to  claim 65 , wherein the hydrogel comprises an acrylamide. 
     
     
         67 . The system according to  claim 65 , wherein the biological specimen is central nervous system tissue. 
     
     
         68 . The system according to  claim 65 , wherein the biological specimen is a biopsy specimen or an autopsy specimen. 
     
     
         69 . The system according to  claim 54 , wherein the cellular components comprise lipids. 
     
     
         70 . The system according to  claim 54 , wherein the imaging is performed using a microscope. 
     
     
         71 . The system according to  claim 70 , wherein the microscope is an optical microscope, a laser microscope, an electron microscope, or a scanning probe microscope.