Post on 19-Mar-2016
description
Detection of Amnionless Protein in
Kidney Sections
AuthorsNatasha TongeDr. Roberta Rivi
Acknowledgements
Dr. Roberta RiviDr. Sat Bhattacharya
Dr. Katia ManovaDr. Betul Altay-Ozer
Harlem Children SocietyElizabeth Lacy Lab
MSKCC
Wax in a Different Context
Tissue sample is embedded in paraffin wax
Wax in a Different Context
Tissue sample embedded in paraffin wax
Sliced with microtome
Wax in a Different Context
Tissue sample embedded in paraffin waxSliced with microtome
Placed in Tissue bath
Wax in a Different Context
Tissue sample embedded in paraffin waxSliced with microtomePlaced in Tissue bath
Mounted on a slide
Wax in a Different Context
Tissue sample embedded in paraffin waxSliced with microtomePlaced in Tissue bathMounted on a slide
Automatically Processed
Immunohistochemistry - a process used to detect proteins in a tissue section.
Antigen - a substance that provokes an immune responseAntibody – the protein complex that binds to antigens
Antibodies in a Different Context
Enzyme - A protein that speeds up a chemical reaction.Substrate - The chemical that induces the chemical reaction
Immunohistochemistry - a process used to detect proteins in a tissue section.Antigen - a substance that provokes an immune responseAntibody – the protein complex that binds to antigens
Antibodies in a Different Context
primary antibody
secondary antibody
substrate
enzyme
Antigen
Enzyme - A protein that speeds up a chemical reaction.Substrate - The chemical that induces the chemical reaction
The Anatomy of the Kidney
Proximal Tubule - close to the glomerulus.Ammnionless - protein located on the proximal tubule.
Cortex - outer region of the kidney. Most likely to find the glomerulus and proximal tubule here.Medula - inner region of the kidney. Lacks ammnionless.
The Kidney in Focus (more or less)
The Kidney in Focus (more or less)
The Past and Future of this Experiment
Tissue samples are sectioned, placed on slides and stained
The staining process, immunohistochemistry, makes use of antibodies to induce a color change
In future experiments the same process will be used topinpoint mutations in abnormal mouse tissue.
To Detect Amnionless in Kidney Sections:
That color change enabled us to see the location of the proximal tubules in the kidney sections.
Bibliography
http://upload.wikimedia.org/wikipedia/en/thumb/b/b0/Kidney_section.jpg/180px-Kidney_section.jpg (altered by Natasha Tonge)http://mskweb3.mskcc.org/MolecularCytology/mccf-final.htm
http://en.wikipedia.org/wiki/Human_hair
Fin