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 - 18 . (canceled) 
   
   
       19 . A method for identifying ultrastructural biomarkers from a biological sample the method comprising:
 providing a substantially thin film of a biological fluid sample to be imaged;   wherein, the sample includes components from one of intracellular organelles or components, extracellular organelles or components, tissue components, and biological fluids;   immersing the sample in a cryogenic fluid so as to cause the sample to vitrify;   transferring the sample onto a stage for placement in a high contrast imager under positive pressure;   generating, from a high contrast imager, an image from a region of the thin film sample for subsequent identification of ultrastructural biomarkers; and   identifying ultrastructural biomarkers in the image.   
   
   
       20 . A method as set forth in  claim 19 , wherein the cellular organelles or components, or tissue components include one of plasma membrane, organelle membranes, basement membrane, extracellular matrix, intercellular organelles, intercellular structures, intracellular membranes, intracellular organelles, cell-cell junctions, cell-cell adhesion, gap junctions, tight junctions, nucleus, nucleolus, nuclear membrane, nuclear pore, chromosomes, chromatin, ribosomes, polyribosomes, monosomes, cellular proteins, cellular protein complexes, cellular protein subunits, extracellular proteins, extracellular protein complexes, extracellular protein subunits, secretory proteins, secreted protein complexes, secretory protein subunits, secreted intracellular or extracellular protein aggregates, golgi, lysosomes, mitochrondria, endosomes, mitochrondrial membranes, peroxisomes, endoplasmic reticulum, mRNA, DNA, tRNA, rRNA, small RNA, proteosomes, vacuoles, intracellular and extracellular vesicles, cavity, and droplets, cellular lipids or carbohydrates, cellular lipid or carbohydrate complexes, cellular lipoproteins, cellular glycoproteins, intracellular and extracellular lipids, extracellular lipoprotein complexes, extracellular lipoprotein subunits, secreted proteins, secreted protein subunits, lipoprotein or glycoprotein aggregates. 
   
   
       21 . A method as set forth in  claim 19 , wherein the biological fluid includes one of blood, mucosa, plasma, serum, cerebral spinal fluid, spinal fluid, joint fluid, urine, saliva, bile, pancreatic fluid, peritoneal fluid, lung fluid, alveolar sac fluid, sinus fluid, lachrymal fluid, nasal mucous and fluid, intrathoracic fluid, gastric fluid, gastrointestinal fluid, ovarian fluid, testicular, prostrate fluid, uterine fluid, cystic fluid, renal fluid, brain fluid, ophthalmic fluid, tear, ear fluid, auditory canal fluid, subcutaneous or muscular fluid. 
   
   
       22 - 41 . (canceled)