microscopy fits needs of researcher, type of specimen being viewed brightfield brightfield darkfield...
TRANSCRIPT
Microscopy fits needs of Microscopy fits needs of researcher, type of researcher, type of
specimen being viewedspecimen being viewed
BrightfieldBrightfield DarkfieldDarkfield DIC/NomarskiDIC/Nomarski UltrastructuralUltrastructural Dimensional microscopy (SEM and Dimensional microscopy (SEM and
Confocal)Confocal) Simple magnification Simple magnification
Resolution Depends Upon Resolution Depends Upon Magnification, Type of Scope, Magnification, Type of Scope, Specimen Fixation, Thickness, Specimen Fixation, Thickness,
StainingStaining Staining types – GeneralStaining types – General
Acid stains (eg., eosin, orange G, aniline Acid stains (eg., eosin, orange G, aniline blue), blue),
Basic stains (Methyl green, methylene blue)Basic stains (Methyl green, methylene blue) Mordants (Hematoxylin, a generally basic Mordants (Hematoxylin, a generally basic
dye, is a mixture of hematoxylin and mordant dye, is a mixture of hematoxylin and mordant (link b/w dye and tissue) generally used in (link b/w dye and tissue) generally used in connection with acidic dyes to provide connection with acidic dyes to provide contrast and staining of cellular componentscontrast and staining of cellular components
Specific tissue component stains (eg., PAS Specific tissue component stains (eg., PAS for carbohydrates)for carbohydrates)
Staining Types - SpecificStaining Types - Specific ImmunohistochemistryImmunohistochemistry Enzyme HistochemistryEnzyme Histochemistry AutoradiographyAutoradiography
Resolution, cont’dResolution, cont’d
Microscope Slide
Primary antibodySecondary Antibody
Protein in tissue
Fluorescent or enzymatic label
Immunohistochemistry
Enzyme in cells of tissue
Substrate
Enzyme Histochemistry
AutoradiographyAutoradiography Incorporation of label into DNAIncorporation of label into DNA
Injection of labelled tracer into animal or cellInjection of labelled tracer into animal or cell Removal of tissue, fixation, sectioningRemoval of tissue, fixation, sectioning Exposure of section to photographic emulsionExposure of section to photographic emulsion Emission of radioactive tracer reacts with Emission of radioactive tracer reacts with
emulsionemulsion Emulsion is developed, silver grains denote Emulsion is developed, silver grains denote
label locationlabel location Counterstain and viewCounterstain and view
Areas of label indicative of cellular Areas of label indicative of cellular process (eg., making DNA, protein) being process (eg., making DNA, protein) being examinedexamined
Guess the Technique
- control + control)
- control + control