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