Compositions and Methods for Quantitative Histology, Calibration of Images in Fluorescence Microscopy, and ddTUNEL Analyses
AssigneeThe Methodist Hospital Research Institute
InventorsMartyn Alun Sharpe, David S. Baskin
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Abstract
Disclosed are compositions and methods for quantitation and calibration of images in fluorescence microscopy. Also provided are tissue phantoms that contain known amount(s) of fluorophore standard(s), as well as components and diagnostic kits containing the same for use in various histological analyses. In certain embodiments, three distinct nucleic-acid based assays provide improvements over conventional TUNEL methods to facilitate precise quantitation of a variety of nucleic acids obtained from a biological sample.
Claims28
A tissue phantom comprising gelatin that is operably linked to at least a first detection moiety.
The article of manufacture of claim 16, comprising a plurality of distinct tissue phantoms, each of which includes a distinct, known amount of the first calibration standard, adapted and configured to facilitate the generation of a standard curve to quantitate one or more selected compounds of interest within a specimen, a sample, a tissue, a cell, or any combination thereof.
A method of modifying a terminal deoxynucleotidyl transferase nick end labeling assay, comprising substituting a 3′ dideoxy UTP substrate for a dUTP substrate under conditions effective to permit quantitation of the —OH 3′ ends present in an assayed biological sample suspected of containing a population of nucleic acids.
A method of quantitating —PO4 3′ ends in a nucleic acid molecule, comprising using calf intestinal alkaline phosphatase to convert the —PO4 3′ ends to —OH 3′ ends, and then assaying the converted —OH 3′ ends using a ddTUNEL assay.
A method of visualizing reactive oxygen species damage within a biological cell, wherein one or more 2,4-dinitrophenyl hydrazine-derivatized oxo-species are localized in the cell by detecting the presence of a labeled anti-DNP antibody.
Dependencies are inferred from the claim text.