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Microscopes

Microscopes are commonly used lab equipment that come in various types such as light, phase contrast, fluorescence, and electron microscopes. The document discusses the components of microscopes like the light source, condenser, specimen stage, objectives, and eyepiece. It also covers objective properties, common microscope problems, cleaning, and servicing methods.

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LenchoGudinaBegna
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157 views21 pages

Microscopes

Microscopes are commonly used lab equipment that come in various types such as light, phase contrast, fluorescence, and electron microscopes. The document discusses the components of microscopes like the light source, condenser, specimen stage, objectives, and eyepiece. It also covers objective properties, common microscope problems, cleaning, and servicing methods.

Uploaded by

LenchoGudinaBegna
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPTX, PDF, TXT or read online on Scribd
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Microscopes

Moisey (2009), Optical Microscope [photograph]. Retrieved from https://commons.wikimedia.org/wiki/File:Optical_microscope_nikon_alphaphot_%2B.jpg


Microscopes:

Microscopes are some of the most commonly
used pieces of lab equipment

Seldom maintained

Medical uses;
 Histology
 Pathology
 Hematology
Types of Microscopes:
•
Compound light
•
Inverted Compound light
•
Phase shift
•
Fluorescence
•
Digital
•
Electron
Arranged in order of commonality
Types of Microscopes
• Light microscopes (magnification ~ 2000X)
– Compound
• Staining needed
– Stereo
• For dissection, etc. (low magnification)
– Phase contrast
• Upright or inverted (for cell culture)
• No staining needed, can see live cells
– Fluorescence (usually with phase contrast) (your limit!!- don’t touch)
– Confocal
• Produces 3-D images (Z-stack)
• Electron microscope (magnification 500,000 X – 5000,000X)
– Scanning, Transmission , Cryo
• Live samples cannot be viewed because of vacuum
• Sample preparation is laborious (except for cryo)
• Cannot transport, dedicated facility needed
Compound Light Microscopes:

•
Monocular/Binocul •
Slide clips
ar •
3-5 Objectives
•
Inverted •
Stage XYZ
•
Light or Reflector •
Locks
•
Filtered
•
Aperature
Optical Path in a Typical Microscope

Wikipedia. “Optical Microscope.” Wikipedia. Retrieved from: https://en.wikipedia.org/wiki/Optical_microscope


Components of a Microscope

• Light source
• Condenser
• Specimen stage
• Fine and coarse focus knobs
• Objective (s)
• Eyepiece (s)
• Camera
Basic Light Microscope
Components

Wikipedia. “Optical Microscope.” Wikipedia. Retrieved from:


https://en.wikipedia.org/wiki/Optical_microscope
Microscope Components
Condenser
A condenser is designed
to collect, control and
concentrate light from the
lamp onto the specimen.
Iris controls the amount
light

Egmason (2011), Condensor specimen [drawing]. Retrieved from


https://commons.wikimedia.org/wiki/File:Condenser_diagram.svg
Microscope Component
Specimen Stage
Anchors the specimen on the slide, dish, etc. and allows
scanning he specimen in X-Y direction using micrometer
screws

Stage

Moisey (2009), Optical Microscope [photograph]. Retrieved from


https://commons.wikimedia.org/wiki/File:Optical_microscope_nikon_alphaphot_
%2B.jpg
Microscope Component
Focus Knob(s)

• Moves the stage or the objective tube up


and down to achieve focus.
• Uses fine micrometer screws.
• All microscopes have a ‘coarse’ and
‘fine’ focus knobs.
• Modern microscopes have them as one
piece as shown
Microscope Component
Objective
Heart of the microscope, determines the magnification of the
image and resolution . Objectives are interchangeable easily to
change the magnification.

WHO. “Learning Unit 6: The Microscope.” From the Publication: Basic Malaria Microscopy: part 1. A Learner’s Guide. WHO: Switzerland (2010), p. 37-44. Retrieved from:
http://apps.who.int/iris/bitstream/10665/44208/1/9789241547826_eng.pdfhttp://apps.who.int/iris/bitstream/10665/44208/1/9789241547826_eng.pdf
Microscope Component
• Magnification
Objective- Properties
– How big the image looks
– As magnification increases, area seen decreases
– Magnification does not always mean better resolution
• Numerical aperture
– How much light is collected from specimen from the objective
– Numerical aperture determines resolution
• Immersion medium
– At high magnification, more light can be captured if immersion
medium such as oil is used
• Working distance
– How close the objective has to be for optimal function
• Depth of focus/field
– The range of distance of the objective from the specimen within which
the image will stay in focus
Objective Properties
Magnification vs. Resolution

High magnification without resolution is not useful;


this is known as ‘empty magnification’
Objective Properties
Oil Immersion
Immersion in media with higher refractive index than air (such as oil)
increase the amount of light collected, hence creates higher numerical
aperture and greater resolution

User Gcg (2009), Immersion Microscopy [photograph]. Retrieved from https://commons.wikimedia.org/wiki/File:Immersion_microscopy.jpg


Objective Properties
Depth of Focus/Field
• The range of distance of the objective from the
specimen within which the image will stay in focus
• Depth of focus decreases with higher numerical
aperture
• The image will go out of focus with small
movements of the objective when depth of focus is
small
• Like f-stop in a camera
Microscope Component
Eyepiece
• Eyepiece is the viewing unit
of a microscope
• Can be mono ocular (one
eye: old microscopes) or
binocular, where both eyes
are used
• Eyepieces add magnification
(usually 10X- 20X) to the
image produced by the
objective
Jastrow (2007), Clave 25mm Eyepiece [photograph]. Retrieved from
https://commons.wikimedia.org/wiki/File:Clave_25mm_eyepiece.jpg
Common Problems:


Dirty Lenses

Dirty Light path

Stiff stage mechanism

Broken or missing parts
Cleaning method:


Clean with 99% alcohol or lens cleaner
ONLY!

Use a chamois or microfibre cloth;
 Cotton batten may be used with care
 Light pressure

If light path is open take care not to
contaminate

Cover microscopes when not in use
Servicing Microscopes:

Clean with 99% alcohol or lens cleaner
ONLY!

Check light path for contamination

Test light range

Check all magnifications for focus and
tracking

Stage must move freely-never force stage
movement with fine/coarse controls

Check locks & slide clamp

Cover when servicing is complete
?
Questions?

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