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51 Cards in this Set
- Front
- Back
It acts as a permanent record of tissues, organs, and specimens. |
Slides |
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What are the info needed to provide in a slide? |
1. Name of the organism then w.m. or e 2. Part of the organism used 3. Type of preparation 4. Date 5. Signed |
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The process of mechanical cutting of plant/animal materials are done by the aid of microtomes |
Microtomy |
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Four basic groups of Microtomy |
1. Rocking microtome 2. Rotary microtome 3. Sledge microtome 4. Freezing microtome |
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generally best for cutting sections thicker than six microns. |
Rocking microtome |
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This microtome is not suitable for embryological specimen specially if the juxtaposition of the cells in critical or when doing serial section |
Rocking microtome |
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More expensive than rocking microtome |
Rotary microtomes |
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enable sections of 5 microns thickness to be cut. |
Rotary microtomes |
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May weigh as much as fifty kilograms, and consists of a heavy base into which aremachined flat tracks on which the chuck sledge slides. |
Sledge Microtome |
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used to cut sections down to two or three microns. |
Sledge Microtome |
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This device usually has a wedge blade |
Freezing microtome |
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Such microtomes are sometimes mounted in chilled cabinets to make working with themeasier |
Freezing microtome |
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now possible to cut frozen sections using freezer aerosols instead of CO2 andfreezing chucks are made which are electrically cooled by means of Peltiercells. |
Freezing microtome |
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three basic cross sectional shapes of knives |
1. wedge 2. plano-concave 3. double concave |
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Knives that is generally,though not exclusively used for cutting frozen sections |
Plane wedge knife |
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Knives are often used for sectioning soft materials such as celloidin embedded tissues. |
plano-concave |
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razors are generally plano-convex. |
Botanical sectioning |
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best knives for the cutting of paraffin blocks |
double concave type |
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Combine histological methods with chemical or biochemical methods, make some compositions of tissue or cell become insoluble, colored or electron-dense, to show those chemical compositions of tissue or cell in situ, such compositions includes protein, amino acid, nucleic acid, lipid and enzymes. |
General Histochemistry |
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¨branch of science concerned with the qualitative and quantitative assessment of chemical compounds in a cell/tissue using stains/dyes and microscopy.¨ |
Histochemistry |
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marriage between bio/chemistry and cyto/histology |
Histochemistry |
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¨He stained starch in plant tissues blue with potassium iodide solution under the light microscope and demonstrated its localization microscopically |
Raspail,1825 |
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he published an essay on microscopic chemistry for the first time |
Raspail,1825 |
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¨first to introduce cell fractionation to analyze nucleic acids in nuclei of leukocytes. ¨ |
Miescher(1874) |
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It is used to analyze nucleic acids in nuclei of leukocytes. |
Cell fractionation |
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¨ microscopic chemistry or chemical microscopy are meant to observe chemical reactions in situ under microscopy. |
Microchemistry |
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¨frequently used to stain tissues in anatomy and pathology during early 20th century |
Aniline dyes |
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published his famous book entitled “Histochemie Animale”, |
Lison(1936) |
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¨He proclaimed Histochemistry to be the new science |
Lison 1936 |
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¨proposed that the field should be designated General Histo-cytochemistry just like we have General Histology. |
Nagata(1995) |
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One of the foremost histochemists in Nigeria |
LateProf. Caxton-Martins |
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classificationof histochemical methods |
1. Chemical methods such as chemical reactions by staining 2. Physical methods such as radiations 3. Biological methods such as immunity |
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¨histology,anatomy, immunology and biochemistry to detect the amount, distribution andlocalization of a specific target within a tissue. |
Immunohistochemistry (IHC) |
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anumbrella term that encompasses all the techniques that are used for thedetection of molecules employing the antigen–antibody reaction |
Immunostaining |
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aspecific use case of immunostaining when the antigen–antibody reaction is usedto study the status of molecules in tissue |
Immunohistochemical staining |
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¨Immunohistochemistryrefers to the evaluation of target antigens in tissues whenthe detection method can be either chromogenic or fluorogenic. |
Immunohistochemistry |
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mayrefer to both the evaluation of target antigens in cells and tissues andspecifically involves the detection of a fluorescent label(fluorophore) |
Immunofluorescence |
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studyof the status of antigens in tissue samples |
immunohistochemistryi |
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immunostainingwhen the sample of interest is cells in culture |
immunocytochemistry |
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Parameters in Performing Immunohistochemistry |
1. tissue 2. target 3. epitope 4. fixation method 5. fixative 6. sample preparation methods 7. sectioning methods 8. pre-processing tissue sections 9. antigen retrieval methods 10. permeabilization 11. blocking buffer 12. detection method 13. primary antibodies 14. secondary antibodies 15. signal amplification 16. labels 17. counterstain 18. mounting 19. multiplexing 20. imaging method 21. controls |
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thesmall three-dimensional surface region of the antigen to which an antibodywould specifically bind. |
epitope |
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Fixativesfall into three categories: |
1. aldehyde 2. alcohols 3. acetone-based fixatives |
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mostcommonly used fixative is |
4%(w/v) paraformaldehyde solution prepared in phosphate buffered saline (PBS). |
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Twocommon methods of sample preparation |
1. formalin-fixed paraffin embedding (FFPE) 2. freezing. |
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Antigenretrieval can be achieved either by the application of |
1. heat(heatinduced epitope retrieval: HIER) 2. enzymatic degradation (proteolytic-induced epitoperetrieval: PIER) |
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surfactants used to achieve permeabilization |
1. TritonX-100 2. Tween-20 3. saponin and digitonin |
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The concentration of the surfactant and the time of incubation are determinedbased on factors such as the |
1. fixative used 2. thickness of the tissue section 3. subcellular localization of the antigen of interest. |
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Commonly used blocking agent |
bovine and serum albumin |
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Primary antibodies may be |
polyclonal or monoclonal |
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Severalsignal amplification strategies that are used includ |
1. avidin-biotincomplex (ABC)method 2. labeledstreptavidin-biotin (LSAB)method 3. tyramide signalamplification(TSA). |
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Labels used to detect the target antibody are attached to the primary or secondary antibodies and may be either _ or _ |
1. fluorogenic 2. chromogenic. |