Jeffrey B. Bindell October 17, 2008 1. Introduction 2. The Optical Microscope Some material taken...
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Transcript of Jeffrey B. Bindell October 17, 2008 1. Introduction 2. The Optical Microscope Some material taken...
Jeffrey B. BindellOctober 17, 2008
1. Introduction2. The Optical Microscope
Some material taken from:http://www.me.ust.hk/~mejswu/MECH343/343om-01principles.pdf
What’s Happening
MORNING SESSION
WELCOME, 8:30 – 9:00 a.m.
Dr. Jeffrey Bindell, University of Central Florida and
Florida High Tech Corridor Council (FHTCC)
Dean John WiencekUniversity of South Florida, College of Engineering
The Optical Microscope, 9:00 – 9:30 a.m.
Dr. Jeffrey Bindell, University of Central Florida & FHTCC
The Scanning Electron Microscope, 9:30 – 10:00 a.m.
Kerry N. Siebein, University of Florida
COFFEE BREAK, 10:00 – 10:15 a.m.
TEM - 10:15 – 11:00 a.m.Dr. Brenda Prenitzer, NanoSpective, Inc
Internet Topics – 11:00 a.m. – 11:30 a.m.Dr. Jeffrey B. Bindell
LUNCH, 11:30 a.m. – 12:15 p.m.
AFTERNOON ROTATION SCHEDULE12:30 p.m. – 3:00 p.m. Sessions 45 Minutes Each
An Introduction to Vacuum and Vacuum CoatingsMr. Robert Tufts & Mr. Richard Everly, University Of South
FloridaNanomaterials and Nanomanufacturing Research Center
(NNRC)
Hands-On Exposure to the Scanning Electron Microscope,
Mr. Joshua Schumacher, University of South Florida (NNRC)
TEM Internet DemoDr. Yusuf Emirov, University of South Florida
(NNRC)
WRAP UP 3:00 – 3:30 p.m.
Jeff Bindell
Introductory Comments on the Optical Microscope
http://www.microscopeworld.com/MSWorld/parts.aspx
What we do with light and now electrons
We look to the outer limits of the universe..
We look at microbes
We look at Virus particles
We look at atoms
We look at POLITICIANS
ObaObamamaMcCaiMcCai
nn
VOTE TWICE
VOTE FOR BOTH!!
We can do all of this imaging becauseWe can bend light with transparent
materials
And we can bend electrons with magnetic fields
The Lens
Incident Ray
Refracted Ray
Transmittedray
Parallel
Notice that if the slab ofglass is this, the incidentand transmitted rays arealmost the continuation
of each other.
Kirkpatrick/Francis – “Physics; A World View”
Some Rays …
Focu
sFocu
s
Stronger
Stronger
bending
bending
toward
toward
the
the
outsid
e of
outsid
e of
the le
ns.
the le
ns.
StrongeStronger r bending
bending toward toward the the outside
outside of the of the lens.lens.
Focu
sFocu
s
Optics ReviewedOptics Reviewed
How does the eye create an image for the brain to understand?
The man on the bicycle appears upside down due to lenses, because lenses invert the image.
OpticsOptics (Continued)(Continued)
The diagram to the right illustrates how the microscope creates an image that the eye can process.
A compound microscope uses a very short focal length objective lens to form a greatly enlarged image.
The Following Slides have The Following Slides have been stolen, without apology, been stolen, without apology,
from Dr. Dempere - UFfrom Dr. Dempere - UF
Total Magnification Example:Total Magnification Example:
If the ocular (eyepiece) lens has a magnification of 10x and the objective lens being used is 100x, the total magnification would be
1,000 x
OpticsOptics (Continued)(Continued)
Images viewed through the eyepiece of compound microscopes will appear
upside-down and backwards.
Slide with the letter “F”
The letter “F” as it appears when
viewed through the eyepiece
F
Important Microscope parts Important Microscope parts include :include :
Ocular lens or eyepiece is used for viewing.
Revolving nosepiece contains objective lenses that are used to magnify the image in combination with the ocular lens.
Total Magnification:Total Magnification: The total magnification of
the specimen being viewed is calculated using the ocular lens multiplied by the objective lens.
For example, if the ocular lens is 10x and the ocular lens is 45x then the total magnification would be 450x.
Other important microscope Other important microscope parts:parts: Stage- the location of the
specimen to be viewed
Clips- utilized in holding the specimen in place
Lamp- typically a light source underneath the stage
Diaphragm- controls the amount of light allowed to pass through the specimen
Focus and Resolution PartsFocus and Resolution Parts
Course-adjustment knob- is the larger of the two knobs. It is used in bringing the object into quick focus.
Fine-adjustment knob- is used for improving the clarity of the image, especially when viewing under high power.
Factors that influence the qualityFactors that influence the quality and size of image: and size of image:
Focal Length Size of sampleType of sampleQuality of
Microscope and lenses
Amount of light on the sample
Quality of sample
Depth of FieldDepth of Field
"Depth of field" refers to the thickness of the plane of focus.
It is the vertical distance (from above to below the focal plane) that yields a useful image.
Depth of Field (continued)Depth of Field (continued)
With a large depth of field, all of the threads can be in focused at the same time.
With a smaller or narrower depth of field, only one thread or a part of one thread can be focused, everything else will be out of focus.
Depth of Field Example:Depth of Field Example:
The series of images show how the depth of field can influence the appearance of an image.
With narrow depth of field, only part of the image is in focus at the same time.
With a large depth of field, the entire image is in focus at the same time.
ResolutionResolution
The amount of detail you can see in an image.
You can enlarge a photograph indefinitely using more powerful lenses, but the image will blur together and be unreadable.
OBSERVATION MODES (A) STAINING
Cell staining is a technique that can be used to better visualize cells and cell components under a microscope. By using different stains, one can preferentially stain certain cell components, such as a nucleus or a cell wall, or the entire cell.
Observation Modes (B)Observation Modes (B)
Phase Contrast- is a type of light microscopy that enhances contrasts of transparent and colorless objects by influencing the optical path of light.
The Phase Contrast microscope is able to show components in a cell or bacteria, which would be very difficult to see in an ordinary light microscope.
Normal Microscope
Phase Contrast
Observation Modes (C)Observation Modes (C)
Polarization- uses two polarizers, one on either side of the specimen, positioned perpendicular to each other so that only light that passes through the specimen reaches the eyepiece.
Estradiol
Ibuprofen
Observation Modes (D)Observation Modes (D)
FluorescenceFluorescence- this technique is used to study specimens, which can be made to fluoresce.
Endothelium Cells
Mitosis in hamster
Advantages of Using Optical Advantages of Using Optical MicroscopesMicroscopes
InexpensiveEasy to learn and
operateMagnification
acceptable up to 2,000 times
Very sharp plane of focus
Small and portable
Standard ImagingGet some high detail images of things of interest – like bugs, microbes (the bigger ones) and pond scum creatures.
MICROSCOPYMICROSCOPY
Presented By The
Florida High Tech Corridor Council
and
techPATH Education Consortium~
This Module was Prepared by
Carmen S. AustinScience Teacher
School District of Hillsborough CountyTampa, Florida
ON THE DISKON THE DISK
Classroom Activities
Experiments
Ideas