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Microscopy and Classification 2
Microscopy
"ow does a lens wor#$
%ptical propertiesrefraction
magnification
resol!tion
distortion
&imitations of a single lens
magnification
ill!mination
aberration 'i#ipedia lin#http://en.wiipedia.or!/wii/"ens#$2%optics$2&
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Microscopy and Classification 3
Some principles of a
compo!nd microscope
%c!lar lens
%b(ective lens
Condenser lens
"ow is magnification calc!lated$
&imitations to a compo!nd
microscope
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Microscopy and Classification '
Speed of light in vac!!m
Speed of light in medi!m
'hat were the #ey advances
in light microscopy$
)* %il immersion lens
+* !sed lenses
* .dvanced optics
"ow do oil immersion andf!sed lenses improve resol!tion$
efractive index /
0orosilicate glass / )12+
.ir / )133
'ater / )1Immersion oil / )12+
!sed lenses combine different types of glass
-- compensate for aberrations
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Microscopy and Classification
.dvanced optics only improve contrast
Theoretical limit of resol!tion
/ 31+ !M
ar# field
manip!lates the
path of light
Phase contrast
and IC
manip!late
phase of light
ar# field
0right field
IC
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Microscopy and Classification
l!orescence Microscopy allows precise
staining of cells and cell!lar components
Standard fl!orescence
manip!lates the
path of light
Confocal -- p!ts everything in foc!s
Standard l!orescence vs Confocal image of Tetrahymena
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Microscopy and Classification *
"ow does electron
microscopy yield
higher resol!tion$
esol!tion
- 5ye + 0.2 mm
- &M + 0.2 ,M
- 5M + 0.2 nM (0.0002 ,M)
'avelengths of
6 of electrons 7 31332 nm
6 visible light 7 223 nm
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Microscopy and Classification %
'hat are types of
electron
microscopy$
Scanning vs Transmission
STM is different
riscoll et al1 )8831
9at!re, 4:, :+;)
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Microscopy and Classification &
0iological classification
0iological taxa
-- M!ddled in the middle
-- 0l!rry at the bottom
0inomial naming
'riting conventions
Staphylococcus aureusStaphylococc!s a!re!s
S. aureus
Common names and strainsPne!moncocc!s
E. coli %)2< "<
Domain: Bacteria
Kingdom: Eubacteria
Phylum: Firmicutes
Class: Bacilli
Order: Bacillales
Family: Staphylococcaceae
Genus: Staphylococcus
Species: aureus
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Microscopy and Classification 10
The Antiquity of Microbes
dam
!ad "em## Stric$land Gillilan
'hat are the domains of life$
Eukarya – Archaea – Bacteria
'hat traits do .rchaea
and 0acteria share$ -- proaryotic cell str,ct,re
-- circ,lar chromosome
'hat traits do .rchaea
and 5!#aria share$ -- ,cleosomes
-- M,ltiple olymerases
-- iosome str,ct,re
Some .rchaea have !ni=!e properties -- enetically distinctie
-- Cell memrane
-- Cell wall str,ct,re
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Microscopy and Classification 11
'hat are the characteristics of
5!#aryotes vs Pro#aryotes$-- all >cell!lar organisms?
0asic similarities$
memrane
!enetic information
iochemistry
istinctive properties of e!#aryotes4 5 e,6 = tr,e 5aryo6 = n,cle,s4 si7e4 or!anelles
mitochondria8 chloroplasts8 etc
4 chromosomes
9s proaryotes 5pre-n,cle,s6
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Microscopy and Classification 12
rchaea E%tremophiles-- some notable ones…
Pyrolobus fumarii -- >ire lobe of the chimney?
"yper-thermophile -- &0C ; 113C (10C)
hydrothermal ents
<train 121:
http://www.!enomenewsnetwor.or!/articles/0%#03/hottes
t.shtml
Picrophilus spp.
5xtreme acidophiles -- p >' (opt 0.*)
?olcanic steam ents
alobacterium spp.
5xtreme halophiles -- 3 ; M salt
@ead seaA !reat salt laeA
<alted foods