Optical microscopy basics

1. OPTICAL MICROSCOPY
THE BASICS

2. OPTICAL MICROSCOPY WAS FIRST CREATED BY
ROBERT HOOKE IN 1665

3. TWO IMPORTANT DEFINITIONS
RESOLUTION: ABILITY TO SEE TWO ADJACENT
OBJECTS SEPARATELY
MAGNIFICATION: RATIO OF IMAGE PRODUCED TO
ACTUAL DIMENSIONS OF SAMPLE

4. OPTICAL LIGHT MICROSCOPY USES
CERVICAL SMEAR
BREAST CANCER IMPRINT
TISSUE FROZEN SECTIONS
CULTURE COVERSLIPS
PARAFFIN WAX EMBEDDED SECTIONS

5. CELLS ARE TRANSPARENT
-STAIN
-LABEL (ENZYME OR FLUORESCENT)
ARE REQUIRED FOR VISUALIZATION

6. CONFOCAL MICROSCOPE
ONLY FOCUSES AND DETECTS LIGHT REACTIONS
THEREFORE CAN CREATE VERY SHARP IMAGES
3D RECONSTRUCTION FUNCTION IS AVAILABLE

7. LIVE CELL IMAGING
ACHIEVED BY GREEN FLUORESCENT PROTEIN