Basic Microscopy – An Overview – October 2005 Protistology Course MBL, Woods Hole, MA.
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Basic Microscopy Basic Microscopy – An Overview – An Overview – – October 2005 October 2005 Protistology Course Protistology Course MBL, Woods Hole, MA MBL, Woods Hole, MA
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Transcript of Basic Microscopy – An Overview – October 2005 Protistology Course MBL, Woods Hole, MA.
Basic Microscopy Basic Microscopy – An Overview –– An Overview –
October 2005October 2005
Protistology CourseProtistology Course
MBL, Woods Hole, MAMBL, Woods Hole, MA
Brief History of the MicroscopeBrief History of the Microscope
What‘s a microscope?What‘s a microscope?
Definition of MagnificationDefinition of Magnification
Conventional Viewing DistanceConventional Viewing Distance
Leeuwenhoek > Compound > Stereo Leeuwenhoek > Compound > Stereo Microscope Microscope
The “Telescope”, a simple detourThe “Telescope”, a simple detour
How to make the specimen visible – How to make the specimen visible – Contrast!Contrast!
Definition of ContrastDefinition of Contrast
Techniques: Techniques: BrightfieldBrightfield PhasePhase DarkfieldDarkfield PolPol DIC (Differential Interference Contrast)DIC (Differential Interference Contrast) FluorescenceFluorescence Optical Sectioning – an expansion of FluorescenceOptical Sectioning – an expansion of Fluorescence
Setting up the Microscope (Lab) Setting up the Microscope (Lab)
Koehler IlluminationKoehler Illumination
Resolution & Empty MagnificationResolution & Empty Magnification
Age
nda
Objects appear to the eye at different magnifications, depending on their distance from the eye. Accommodation (lens) will make it possible.
MB ~ 2x MA
A B
What is “Magnification”?
Conventional Viewing Distance
250 mm
1x
?
“Magnification” 1x
f = 250 mm
1x
1x
Magnification via Single Lens
f = 250 mm
1x
Example: f=50mm
5x
Magnifying Glass (Loupe)Lensf
mmM
250
The simple microscope
Leeuwenhoek Microscope
The -corrected Compound Microscope
Eyepiece
Tube
Objective f250mm
250mm
f
f250mm
M
EyepieceObjectiveMicroscope Compound M MM
Tube lens
(Zeiss: f=164.5mm)
Objective
Eyepiece
EyepieceObjective
Tube
f250mm
ff
M
Q: What happens if we take the objective away?
Tube lens
(Zeiss: f=164.5mm)
Objective
Eyepiece
Eyepiece
Tube
f
fTelescopeM
Eyepiece
Tube
f250mm
250mm
fM
Answer: We have created a “Telescope”
∞
∞
AxioImager Upright Research Microscope
Axiovert 200 Inverted Research Microscope
The basic light microscope types
Upright microscope.
Inverted microscope
Illumination via Transmitted Light
The specimen must be transparent !
Upright microscope.
Inverted microscope
Illumination via “Reflected” (Incident) Light
Eg. Fluorescence, Opaque Samples
Upright microscope.
Inverted microscope
Upright microscope.
Inverted microscope
Mixed Illumination
Other Ways to Illuminate
Reflectors
Ring Lights
Fiber Optics
LED’s
Etc.
Which Microscope types are typically illuminated this way?
Reflectors
Ring Lights
Fiber Optics
LED’s
Etc.
“Couldn’t one build a microscope for both eyes, and thereby generate spatial images?”
Question addressed to Ernst Abbe in 1896
by Horatio S. Greenough
1896: Drawing by Horatio S. Greenough
1897 – the first Stereo Microscope in the world, built by Zeiss, according to the “Greenough” principle
Greenough Type
Telescope Type Introduced first by Zeiss - 1946
Introduced first by Zeiss - 1897
Greenough Stereo Microscopes
SteMi DV4
Greenough Stereo Microscopes
SteMi 2000
(2000-C, 2000-CS)
Research Stereo Microscopes
SteREO Discovery V12 SteREO Lumar V12
• Short BreakShort Break
