Detection of Amnionless Protein in Kidney Sections

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Presentation transcript:

Detection of Amnionless Protein in Kidney Sections Acknowledgements as well. Maybe an animation to replace the authors. A bump Authors Natasha Tonge Dr. Roberta Rivi

Harlem Children Society Acknowledgements Dr. Roberta Rivi Dr. Sat Bhattacharya Dr. Katia Manova Dr. Betul Altay-Ozer Harlem Children Society Elizabeth Lacy Lab MSKCC The purpose of the study is. We do that going through this process : the steps of which are written on the star or circle. The summary should be basic bullet points, a literal couple of words that will be easy to remember though the talk about them will be longer. This is an experiment in histology. Using such and such we compared a wild type mouse kidney to a mutant exposed to enu and try to detect the presence of megalin in its kidney.

Wax in a Different Context Tissue sample is embedded in paraffin wax Is it possible to have something point to each word as they are defined? The talk part should give a brief “history” while the written should be few words. I’d like to review some terms before we proceed. Can the previous definition be made smaller? Not unless Flash is used it seems.

Wax in a Different Context Tissue sample embedded in paraffin wax Sliced with microtome Sliced with microtome - standard 5-12 micrometers. These samples are 8 micrometers thin. Diameter of human hair at its smallest is about 17

Wax in a Different Context Tissue sample embedded in paraffin wax Sliced with microtome Placed in Tissue bath Is it possible to have something point to each word as they are defined? The talk part should give a brief “history” while the written should be few words. I’d like to review some terms before we proceed. Can the previous definition be made smaller? Not unless Flash is used it seems.

Wax in a Different Context Tissue sample embedded in paraffin wax Sliced with microtome Placed in Tissue bath Mounted on a slide Is it possible to have something point to each word as they are defined? The talk part should give a brief “history” while the written should be few words. I’d like to review some terms before we proceed. Can the previous definition be made smaller? Not unless Flash is used it seems.

Wax in a Different Context Tissue sample embedded in paraffin wax Sliced with microtome Placed in Tissue bath Mounted on a slide Automatically Processed Is it possible to have something point to each word as they are defined? The talk part should give a brief “history” while the written should be few words. I’d like to review some terms before we proceed. Can the previous definition be made smaller? Not unless Flash is used it seems.

Antibodies in a Different Context Immunohistochemistry - a process used to detect proteins in a tissue section. Antigen - a substance that provokes an immune response Antibody – the protein complex that binds to antigens Enzyme - A protein that speeds up a chemical reaction. Start with a human cell. Then add an antibody from another animal . Say a rabbit. Then add an antibody from yet another animal. Like a goat with an enzyme attached to it. When that enzyme comes in contact with a substrate a color change is induced. In the case of this experiment, our initial antigen was a short protein chain in the proximal tubule of a mouse kidney. Discovery Substrate - The chemical that induces the chemical reaction

Antibodies in a Different Context Immunohistochemistry - a process used to detect proteins in a tissue section. Antigen - a substance that provokes an immune response Antibody – the protein complex that binds to antigens Enzyme - A protein that speeds up a chemical reaction. Substrate - The chemical that induces the chemical reaction primary antibody Start with a human cell. Then add an antibody from another animal . Say a rabbit. Then add an antibody from yet another animal. Like a goat with an enzyme attached to it. When that enzyme comes in contact with a substrate a color change is induced. In the case of this experiment, our initial antigen was a short protein chain in the proximal tubule of a mouse kidney. Discovery secondary antibody enzyme substrate Antigen

The Anatomy of the Kidney Ammnionless - protein located on the proximal tubule. Proximal Tubule - close to the glomerulus. 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 mouse kidney was the antigen in this case.

The Kidney in Focus (more or less)

The Kidney in Focus (more or less)

The Past and Future of this Experiment To Detect Amnionless in Kidney Sections: Tissue samples are sectioned, placed on slides and stained The staining process, immunohistochemistry, makes use of antibodies to induce a color change That color change enabled us to see the location of the proximal tubules in the kidney sections. In future experiments the same process will be used to pinpoint mutations in abnormal mouse tissue.

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