Lecture 1 - Fall 2007

Resolution vs Magnification Is Bigger Always Better?

Magnification

Distinguishing Detail    



Light Usage Stain Usage Detection of Specific Proteins Detection of Specific Messenger RNA Detection of Polymorphisms in Genes

Resolution Depends Upon Magnification, Type of Scope, Specimen Fixation, Thickness, Staining 

Staining types – General • Acid stains (eg., eosin, orange G, aniline 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 • Specific tissue component stains (eg.,

Resolution, cont’d 

Staining Types - Specific • Immunohistochemistry • Enzyme Histochemistry • Autoradiography

Light

Sherwin, J. R. A. et al. Mol Endocrinol 2004;18:2185-2195

Stain

H&E

Picro Mallory

Specificity

Secondary Antibody

Microscope Slide

Protein in tissue

Primary Antibody

Fatunase 2003

Actin staining of monkey fibroblast cell

Actin staining of microfilaments (red fluorescence)

In Situ Hybridization

In Situ PCR + Chlamydia Primer

- Chlamydia Primer

Guess The Technique