MICROSCOPES Light (visible) Fluorescent U-V Electron Monocular 4/17/2017 MICROSCOPES “illumination” Light (visible) Fluorescent U-V Electron eyepieces Monocular Binocular Resolving Power N.A. affects Wavelength affects specimen background Bright field Dark field MICROSCOPES Microscopes

PARTS & TERMS Lenses (ocular, objective) Total Magnification Condenser 4/17/2017 PARTS & TERMS Lenses (ocular, objective) Total Magnification ocular mag. X objective mag. Condenser Diaphram MICROSCOPES Microscopes

PARTS & TERMS Numerical Aperture (N.A.) 4/17/2017 PARTS & TERMS Numerical Aperture (N.A.) optical characteristic of a lens N.A. = i sinq N.A. increases with magnification MICROSCOPES Microscopes

[ RP of human eye ~ 0.2mm = ______m ] 4/17/2017 PARTS & TERMS Resolving Power (R.P.) size of the smallest discernable detail minimum distance between objects so that they are able to be distinguished as separate (l in nm) R.P. = wavelength of illumination (2)N.A. MICROSCOPES [ RP of human eye ~ 0.2mm = ______m ] Microscopes

Resolving Power (R.P.) l = 650nm, NA = 0.25 4/17/2017 Resolving Power (R.P.) l = 650nm, NA = 0.25 R.P. = l/2NA = _____nm = _____ m 1300 1.3 l = 650nm, NA = 1.25 R.P. = l/2NA = _____nm = _____ m 260 0.260 l = 450nm, NA = 1.25 R.P. = l/2NA = _____nm = _____ m 180 0.180 MICROSCOPES Microscopes

Larger or Smaller R.P. = BETTER ? 4/17/2017 Resolving Power (R.P.) R.P. = l/2NA Larger or Smaller R.P. = BETTER ? R.P. improves as N.A. increases or decreases? R.P. improves as l increases or decreases? MICROSCOPES Microscopes

4/17/2017 Types of Scopes MICROSCOPES Microscopes

4/17/2017 Types of Scopes MICROSCOPES Microscopes

Types of Scopes Compound Light Bright or Dark field 4/17/2017 Types of Scopes Compound Light Bright or Dark field Use blue light or blue filter for shorter l R.P. ~ 0.2m MICROSCOPES Microscopes

4/17/2017 Microscopes

Types of Scopes Fluorescent Dark field compound light microscope 4/17/2017 Types of Scopes Fluorescent Dark field compound light microscope Uses U-V for side illumination of specimen, fluorescent parts or dyes give off visible light that is viewed R.P. ~ 0.2m MICROSCOPES Microscopes

Types of Scopes UltraViolet (U-V) Uses U-V as illumination (shorter l) 4/17/2017 Types of Scopes UltraViolet (U-V) Uses U-V as illumination (shorter l) Image recorded then viewed… still or video camera image (should NOT look directly at U-V!) Special lens material (quartz), glass absorbs U-V R.P. ~ 0.15m MICROSCOPES Microscopes

Types of Scopes Electron Microscopes 4/17/2017 Types of Scopes Electron Microscopes Use electron beam for illumination, magnets for “lenses”, video to view Transmission Electron Microscope (TEM) beam passes thru, view internal structure Scanning Electron Microscope (SEM) beam reflects, see external structure, texture MICROSCOPES Microscopes

4/17/2017 Microscopes

Types of Scopes Transmission Electron Microscope (TEM) MICROSCOPES 4/17/2017 Types of Scopes Transmission Electron Microscope (TEM) MICROSCOPES Microscopes

Types of Scopes Transmission Electron Microscope (TEM) 4/17/2017 Types of Scopes Transmission Electron Microscope (TEM) Special specimen preparation (vacuum) Working magnification: whole bacteria….. 8,000x - 10,000x thin section, viruses…. 30,000x - 40,000x Maximum magnification ~100,000x R.P. ~0.001m = ______nm MICROSCOPES Microscopes

4/17/2017 rickettsia herpes simplex Microscopes

Types of Scopes Scanning Electron Microscope (SEM) MICROSCOPES 4/17/2017 Types of Scopes Scanning Electron Microscope (SEM) MICROSCOPES Microscopes

Types of Scopes Scanning Electron Microscope (SEM) 4/17/2017 Types of Scopes Scanning Electron Microscope (SEM) Special specimen preparation (vacuum & conductor) Working magnification: 14,000x - 50,000x Maximum magnification ~130,000x R.P. ~0.01m = ______nm MICROSCOPES Microscopes

4/17/2017 Q fever (rickettsia) Image enhancement Microscopes