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!!

OPTICALMICROSCOPES

SCANNING ELECTRONMICROSCOPES

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

Simple Lens

Kirkpatrick/Francis – “Physics; A World View”

SPHERICAL SURFACE

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

Major Parts of the Optical Microscope

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

Getting in FocusGetting in Focus

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.

ResolutionResolution

Higher resolution Lower resolution

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

•Confocal Microscopy

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

Expensive Cheap**

** If you can find one.

Standard ImagingGet some high detail images of things of interest – like bugs, microbes (the bigger ones) and pond scum creatures.

AMAZINGMICROLIFE

Created by Shari Waters for

The Orlando Science Center

GREEN PROTISTS

Cosmarium

ROTIFERS

Testudinella

MOVIE

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