Drosophila larval hemisegment, 20x

      Spring 2004 

Capsella  ursa-pastoris, 25x 

Higley Room 214 

Office Hours  M,W 10-noon, T2:30-3:30

     PBX 5398

edwardsr@kenyon.edu

Text: Barbara Foster. 1997. Optimizing Light Microscopy for Biological and Clinical Laboratories.  ASCLS.  Kendell/Hunt Publishers.
 

This laboratory is a practical course (not an experimental course) in light microscopy.  It is designed to give you a theoretical basis in and a "taste" of the power and limitations of light microcopy as an investigative tool. 

Microscopy and image analysis protocols require time, diligence/patience, respect, and care.  Some protocols may require weeks of work prior to observation.  You must invoke patience and utilize great care to maximize the chances for successful outcomes.  People proceed at different rates and returning to lab during the week to complete exercises is encouraged, rather than racing through them and garnering less.

The pedagogy of the class is one of cooperative and collaborative learning.  Discussion/queries/advice shared amongst us in the class is highly encouraged.  I thank this and every class for informing changes that make this course a more effective and fruitful one.

1. Attendance

2. Inquisitive and helpful attitude, diligence and perseverance.

3. Respect and care of the equipment (its all very expensive)

4. Project reports, both oral and annotated PowerPoint, as requested. (library and lab work)

5.  2 laboratory exam/practical.

   Human sperm, 1500x
 

Mouse tongue epithelia,  170x

KENYON COLLEGE 

TEXT:   Barbara Foster. 1997. Optimizing Light Microscopy for Biological and Clinical Laboratories.  ASCLS.  Kendell/Hunt Publishers.

This is the first offering of this specialized laboratory.  Due to facility and staff reasons, the course will be a “rough draft”, something in the making that we will shape together.

Thus the syllabus is not written in stone.  We may not complete all; we may go further.  We may head into other avenues of microscopic inquiry.  Students and the instructor will present materials on aspects of microscopy and analysis throughout the course.

Date     

Topic

¨Text

¨Lab                      ¨Links

January  16

Early history of microscopy:  The single lens microscope

pp 141-142

Make single lens microscopes  

http://www.microscopy.fsu.edu/

primer/museum/index.html

23
 

The dissecting and compound/inverted microscopes: Parts, use, care (Olympus CH2 video).

Kohler alignment/illumination

Intro

Ch. 1

Observe organism’s cellular structure and its development with dissecting and compound microscopes.  Submit drawings and description of organism.

http://www.microscopyu.com/tutorials/java/kohler/index.html

 30

                   

The nature of light and lenses

Brightfield

Student presentations on how the eye “sees”:

visualization, retina structure and thresholds, optic nerve and brain decoding, color and contrast sensitivity

Ch. 2

Ch. 8

Kohler handout

Quantification: determination of actual magnification and making cellular measurements (handout)

http://www.microscopyu.com/articles/ergonomics/ergointro.html

eronomics of microscopy viewing

http://www.microscopyu.com/articles/formulas/measurehome.html

quantification

http://www.microscopyu.com/tutorials/java/components/characteristicrays/index.html

light and lenses

http://micro.magnet.fsu.edu/primer/java/microscopy/immersion/index.html

oil immersion

   February  6    

Nature of light and lenses cont.

Optimizing contrast techniques

Chp 3

Chp 4

Handout on Airy disk

http://micro.magnet.fsu.edu/primer/lightandcolor/diffractionintro.html

http://micro.magnet.fsu.edu/primer/java/diffraction/basicdiffraction/index.html

Diffraction and Airy disk

http://micro.magnet.fsu.edu/primer/java/imageformation/rayleighdisks/index.html

http://micro.magnet.fsu.edu/primer/anatomy/numaperture.html

Airy disk

http://www.microscopyu.com/articles/formulas/formulasindex.html

concepts and formulas

 13

Optimizing contrast techniques

Darkfield

Ch. 5

Lab test

http://micro.magnet.fsu.edu/primer/techniques/contrast.html

contrast techniques

http://micro.magnet.fsu.edu/primer/techniques/contrast.html

darkfield

http://micro.magnet.fsu.edu/primer/java/rheinberg/index.html

Rheinberg

http://micro.magnet.fsu.edu/primer/techniques/oblique/obliquehome.html

oblique illumination

 20

Phase contrast microscopy

 Ch. 6
 Ch. 5

http://micro.magnet.fsu.edu/primer/techniques/phasecontrast/phaseindex.html

http://www.microscopyu.com/tutorials/java/phasecontrast/microscopealignment/index.html

Phase Contrast

http://micro.magnet.fsu.edu/primer/techniques/hoffman/hoffmanindex.html

Hoffman Modulation Contrast

27
 

Polarized light microscopy

Ch. 7:

pp. 78-88

Convert a compound brightfield microscope to polarizing.  Handout: polarized light and biological organization

http://micro.magnet.fsu.edu/primer/techniques/polarized/polarizedhome.html

http://www.microscopyu.com/articles/polarized/polarizedintro.html

March   

   6-20

Spring Break

20
  

Histology: the art of specimen preparation and slide making

Handout

  

27

Differential interference microscopy: DIC Histology cont. (sectioning with Spencer Microtome)

Ch. 7: pp.89-95

http://micro.magnet.fsu.edu/primer/techniques/dic/dichome.html

http://www.microscopyu.com/tutorials/java/dic/dicalignment/index.html

http://www.microscopyu.com/galleries/dicphasecontrast/index.html

compares phase contrast and DIC images

April 3
 

Fluorescence microscopy and image capture

Ch. 7: pp. 72-78

Ch. 9

http://micro.magnet.fsu.edu/primer/techniques/fluorescence/fluorhome.html

http://www.microscopyu.com/articles/fluorescence/filtercubes/filterindex.html

http://www.microscopyu.com/tutorials/java/arclamp/index.html

Fluorescence

http://www.microscopyu.com/articles/photomicrography/digital/index.html

Digital imaging

      10

Limitations to light?  Electron microscopy Histology cont.

Ch. 10: pp 137-140

http://www.unl.edu/CMRAcfem/em.htm

17
 

Limitations to light microscopy? Viewing individual molecules in action with the light microscope.

Lab test

http://www.microscopyu.com/articles/general/innovations.html

innovations

http://www.microscopyu.com/articles/interferometry/twobeam.html

interferometry...interference microscopy innovation

    24      

Deconvolution and Confocal microscopy: 3D imaging

Ch. 10: pp. 133-134

Getting a stereo image from a compound microscope

Resolving and quantifying in 3D (if system software is working)

http://micro.magnet.fsu.edu/primer/techniques/confocal/index.html

http://www.microscopyu.com/articles/confocal/index.html

confocal

http://micro.magnet.fsu.edu/primer/anatomy/stereohome.html

http://www.microscopyu.com/articles/stereomicroscopy/index.html

stereomicroscopy

     May 1       

Student Presentations: Literature examples of how light microscopies are used to advance biological questions/research.