Systems and Methods of Identifying Biomarkers for Subsequent Screening and Monitoring of Diseases

Abstract

A system for generating an image of ultrastructural biomarkers from a biological sample is provided. The system includes a grid onto which a sample to be imaged may be placed and a cryogenic reservoir into which the grid and sample may be immersed for vitrification of the sample. The system also includes a stage onto which the grid and sample may be situated for subsequent imaging in a high contrast imager to permit identification of ultrastructural biomarkers therein. A method for generating an image of ultrastructural biomarkers from a biological sample is also provided. The generated image of ultrastructural biomarkers may be used subsequently for screening and monitoring diseases, evaluating drug and therapeutic efficacy, and assessing risks associated with a drug or therapeutic candidate, among other things.

1 . A system for identifying ultrastructural biomarkers from a biological sample, the system comprising:
 a grid onto which a biological sample to be imaged may be placed;   a cryogenic reservoir into which the perforated grid and sample may be immersed for vitrification of the sample;   a stage, provided with a temperature substantially similar to the cryogenic reservoir, and onto which the grid and sample may be situated for subsequent imaging; and   a high contrast imager designed to receive the stage with the grid for imaging a region of the thin sample for subsequent identification of ultrastructural biomarkers.   
   
   
       2 . A system as set forth in  claim 1 , wherein the grid includes a plurality of holes across which the sample may extend, so as to enhance generation of a thin film thereacross. 
   
   
       3 . A system as set forth in  claim 1 , wherein the grid further includes a first plate onto which the grid may be positioned and a second plate for placement onto the grid. 
   
   
       4 . A system as set forth in  claim 3 , wherein the first and second plates act to spread the sample across the grid, so as to generate certain portions that can be substantially thinner in thickness than others across the grid. 
   
   
       5 . A system as set forth in  claim 3 , further including cryogenically cooled forceps to permit separation of the grid from the first and second plates. 
   
   
       6 . A system as set forth in  claim 1 , wherein the cryogenic reservoir includes an inner chamber for accommodating a first cryogenic fluid, and an outer chamber situated about the inner chamber for accommodating a second cryogenic fluid. 
   
   
       7 . A system as set forth in  claim 6 , wherein the first and second cryogenic fluids are different and the presence of the second cryogenic fluid in the outer chamber helps to maintain the first cryogenic fluid substantially close to its melting point. 
   
   
       8 . A system as set forth in  claim 6 , wherein the cryogenic reservoir further includes a grid holder positioned within the outer chamber for placement of the grid thereon prior to transference onto the stage. 
   
   
       9 . A system as set forth in  claim 1 , wherein the stage includes a container for accommodating a cryogenic fluid, and an arm extending from the container for placement of the grid thereon, the arm having a channel along which cryogenic fluid from the container may flow toward the grid, so as to maintain the temperature of the grid substantially similar to that of the cryogenic reservoir. 
   
   
       10 . A system as set forth in  claim 1 , wherein the high contrast imager can generate substantially artifact-free images in the absence of contrasting agents. 
   
   
       11 . A system as set forth in  claim 1 , further including a positive pressure environment within which the grid may be transferred onto the stage and into the high contrast imager, so as to maintain the integrity of the sample and to minimize risks of contamination of the sample. 
   
   
       12 . A system as set forth in  claim 1 , wherein the biomarkers include components from one of intracellular organelles or components, extracellular organelles or components, tissue components, and biological fluids. 
   
   
       13 . A system as set forth in  claim 12 , wherein the identified ultrastructural organelles or components, extracellular organelles or components can be used to evaluate, determine, or predict drug or therapeutic efficacy, patient response or response rate, or clinical trial participant response or response rate. 
   
   
       14 . A system as set forth in  claim 12 , wherein the identified ultrastructural biomarkers, when compared to ultrastructural biomarkers from healthy or control intracellular organelles or components, extracellular organelles or components, tissue components, and biological fluid can act to assess risks for adverse events, toxicity, or serious adverse events associated with drug or therapeutic candidates in preclinical development, in animal models, or in clinical development, as well as risks for drug attrition in preclinical development, animal models, clinical development or in marketed drugs. 
   
   
       15 - 41 . (canceled)