Contents

Appendix 1

Advanced Project

We have seen that the area in which the inexpensive microscope is most seriously lacking is that of substage illumination. For the microscope with mirror, one can simply purchase a better illuminator, although this will not make up for the lack of a condenser or a rudimentary fixed condenser. Inexpensive microscopes with built in lights may seem even less flexible. However, the following project can provide perfect Köhler illumination for most microscopes. The project is actually a vertical light table built under your microscope. Light tables are used by optics designers for experimentation. Not only will the table allow you to set up Köhler illumination, it will also encourage advanced experimentation with other illumination setups.

The figures depict the illumination table with a microscope sitting on top.

All of the photomicrographs used for this book were taken through a microscope sitting on such a light table. The microscope has standard, non-plan, achromatic optics.

We have separated the project to an appendix because your microscope's warranty will probably be voided by it, and because more skills, tools, and time are required than in earlier projects. Expect to spend about sixteen hours on the carpentry and an additional eight hours or so fiddling with the adjustment of the optical components before the right combination of distances and angles is achieved. The figure gives the dimensions of the table, although you will probably want to modify them for your height and your microscope.

These figures show how the supports for the optical components are attached to the table.

The next figures give the dimensions of these support components. The component that directly supports the optical parts (OC) is shown bolted above the L-shaped support component (LC), but it can be bolted below as well. Some situations may also require that it be made of thinner stock. LC can also be flipped upside down where necessary. The back support component (BC) is used to attach LC to the table.

Any of the members can be designed with extra bolt holes to suit your needs. Except for BC, which needs a bolt at either end, a single bolt will probably be sufficient for attachment of all supports. Small bolts (¼") are used with oversized washers in holes that are larger than the bolt shaft (½"). This allows fine adjustment of optical components. Place a washer that is the right size for the bolt next to the bolt head or nut and the oversized washer next to the wood.

In some cases you may want to provide a member with a slot instead of a hole to allow a wider range of adjustment. A router or jointer makes the best slots, but you can improvise by drlling multiple holes and smoothing the inside of the slot with a coping saw and sand paper.

The final figure shows how an optical component fits into OCM. A hole slightly smaller in diameter than the optical component is drilled at the optical axis and a hole the size of the component is drilled part way through the board. The component rests in this. If you do not have a hole saw the exact size of the component, you will still be able to shim it into a slightly larger hole using cardboard or the like.

Cost:

Costs should run between $150 and $200, excluding tools. Costs can be cut by salvaging old lenses and camera diaphragms, and renting tools is an option.

Tools:

Drill press or, at the minimum,

a large power hand drill;

Complete set of hole saws for the drill;

Set of wood boring drill bits;

Band saw, if rounded corners are desired (a sabre saw can be substituted);

Table saw (a power hand saw or a hand saw with a miter box can be substituted);

Screw drivers and wrenches;

Sandpaper.

Materials

Clear grade 1x4 (about 24') and 1x8 lumber (about 12');

Clear grade 1/2x4 lumber for thinner OC components (small amount -- 3' should be sufficient);

Wood screws;

Wood glue;

1/4 inch bolts, nuts, and Oversize washers.

Optical Components

Ceramic light fixture, standard size;

50 - 75 watt halogen bulb for a standard fixture (either clear or with small pyramids on the

front of the bulb);

Disk or sheet of heat absorbing glass (you don't want that big halogen bulb to damage your objectives);

Large positive lens with focal length between five and ten inches;

Small positive lens with focal length of about two inches (plano convex preferred -- this lens will not be needed if a focusing condenser is already present on

the microscope. See focal length in glossary for method of measurement.);

One or two iris diaphragms, depending on how many your microscope already has;

Ground glass disk or sheet (to allow low power work, where Köhler illumination does not apply);

Light dimmer, as described in earlier project.

Cut the pieces, drill all the holes, and predrill screw holes before assembling the table with wood screws and glue. The light dimmer can be built into the table if you wish, or can be left on an extension cord.

Remove the plate on the bottom of the microscope from the bottom of the base and remove the light and wiring. Replace the rubber feet without the plate. Unscrew the light housing from the top of the microscope base. You should now have an open hole in the microscope base that light from below can pass through.

Review the box on setting up Köhler illumination on page 64. If the microscope has a fixed condenser, the small lens must be placed near it, with a diaphragm just below. The flat side of the lens is placed face down. A filed diaphragm must be placed between the aperture diaphragm and this lens. The lighting must be arranged so that a sharply focused image of either the filament (with a clear bulb) or the pyramids on the front of the bulb fills the bottom of the closed aperture iris. It may help to place a piece of white paper in the aperture iris so the image will be easier to see.

Set the large lens and aperture diaphragm in position, then move the light up and down below the large lens until the image of the light is focused on the bottom of the aperture diaphragm. If the image is too large, move the lens further away from the aperture and repeat; if the image is too small, move the lens closer to the aperture and repeat.

The field diaphragm must also be in focus when viewed through the microscope. This is usually done by moving the small lens up and down, although you have the option of moving the field diaphragm as well. Sometimes, a good location for the field diaphragm is setting on the top of the hole left in the base of the microscope after the built-in lighting components are removed. It all depends on the lenses you have available. It may be necessary to reset the field and aperture diaphragm distances and both lens distances several times, since a movement in any one may cause a change in the others. You may also have to make modifications to some of the support components to get just the right distances between optical components and to get the optical components set correctly in their mounts.

Center all elements by adjusting the bolt that attaches the OC to the LC components. The microscope can also be moved around above the hole in the board that supports it. You may also have to shim up two of the microscope feet since the optical axis of the microscope may not be perfectly vertical.

The heat absorbing glass can be placed just above the light. However, any filter that is not smooth glass (i.e. a gelatin filter or the like) should be placed above the heat absorber and very near the bottom of the aperture diaphragm, where it will interfere the least with the focused image of the light.

To prevent glare, paint the inside of the light table matte black.

You will be surprised at how much light gets to the objective without heating it or the specimen when using a large halogen bulb and a heat absorbing filter. Heat is, however, created in different parts of the system.

WARNING:
1) The light fixture can be screwed directly to an OC support component, but be sure to use a ceramic fixture, since a large halogen bulb creates heat.

2) The OC support for the heat absorbing glass may be best placed near the bulb. To prevent fire, coat the bottom of the OC with aluminum foil to reflect some of the heat away from the wood, or fashion this OC from some non-flammable material. If you cannot place your hand comfortably on the bottom of this OC after the light has been left at high intensity for a while, move the OC further from the light. Dim the light when you walk away from the table.

3) A clear container with a smooth bottom and two inches of water can be substituted for the heat absorbing glass.

WARNING: To prevent electrical shock, never let water spill on the electrical components. Do not use more water or a larger container than necessary. Do not move the table without first unplugging the electrical components and removing the water.

4) Younger microscopists should obtain permission and supervision before undertaking this project.

CAUTION: Clean all sawdust from the table and the environment before bringing the microscope into the room where the project is being built.

Contents

Appendix #2: Suppliers

Baxter Diagnostics Inc. Scientific Products
1430 Waukegan Road
McGaw Park, IL 70085-6787
(Microscopes and scientific products)

Biology Store, The
P.O.Box 2691
Escondido, CA 92033
619-745-1445
(Microscopes, living and preserved biological materials)

Chiu Technical Corp.
252 Indian Head Rd.
Kings Park, N.Y. 11754
516-544-0606
(Microscope illuminators)

Connecticut Valley Biological
82 Valley Rd., P.O. Box 326
Southampton, MA 01073
413-527-4030
(Microscopes, scientific products of all kinds)

Delta Biologicals (a division of Southwestern ScientificCo.)
P.O. Box 26666
Tucson, AZ 85726-6666
800-821-2502
(Preserved biological materials)

Dolan -- Jenner Industries, Inc.
Blueberry Hill Industrial Park
Box 1020
Woburn, MA 01801-9982
800-833-4237
(Microscope illuminators)

Edmund Scientific Company
101 East Glouchester Pike
Barrington, NJ 08007-1380
609-547-8880
(Microscopes, microscope components, other optical components, a wide selection of scientific products of all kinds)

EMD (a division of Fisher Scientific)
4901 W. LeMoyne St.
Chicago, IL 60651
800-955-1177
(Microscopes, scientific products of all kinds)

Modulation Optics, Inc.
100 Forest Drive at East Hills
Greenvale, NY 11548
516-484-8882
(Modulation contrast condensers and objectives)

Nasco Science
901 Janesville Ave.
Fort Atkinson, WI 53538-0901
414-563-2446
800-558-9595
(Microscopes, scientific products of all kinds)

Southern Precision Instrument Co.
P.O. Box 2078
San Antonio, TX 78297
512-224-5801
(Microscopes, microscope components, other scientific instruments)

Stangert Corporation
240 Long Beach Rd., P.O. Box E
Island Park, NY 11558-0504
516-432-4277
(Microscopes)

Start International
3361 Boyington, Suite 120
Carrollton, TX 75006
800-527-1809
(Industrial optical inspection and measuring devices)

Unitron Inc., Instrument Group
170 Wilbur Place
P.O. Box 469
Bohemia, NY 11716
516-589-6666
(Microscopes, polarizing microscopes)

Contents

References

CHAPTER 1

Bradbury, S. The Evolution of the Microscope, Oxford: Pergamon Press, 1967.

Bureau of Naval Personnel. Basic Optics and Optical Instruments, New York: Dover Publications, 1969. (Reprint of Opticalman 3 & 2, NAVPERS 10205, 1969.)

Clay, Reginald S. and Court, Thomas H., The History of the Microscope, London: Charles Griffin and Company, 1932.

Dobell, Clifford. Antony van Leeuwenhoek and His 'Little Animals': Being Some Account of the Father of Protozoology and Bacteriology and His Multifarious Discoveries in the Disciplines, New York: Russell and Russell, 1958.

Hooke, Robert. Micrographia, or Some Physiological Descriptions of Minute Bodies Made by Magnifying Glasses with Observations and Inquiries Thereupon, New York: Dover, 1961. (Facsimile reproduction of the first edition, Royal Society, 1665.)

Jacker, Corinne. Window on the Unknown: A History of the Microscope, New York: Charles Scribner and Sons, 1966.

Johnson, B. K. Optics and Optical Instruments: An Introduction with Special Reference to Practical Applications, New York: Dover, 1960.

Pedrotti, Frank L. and Pedrotti, Leno S. Introduction to Optics, Englewood Cliffs, N.J.: Prentice Hall, Inc., 1987.

Phin, John. Practical Hints on the Selection and Use of the Microscope, Philadelphia: David McKay, 1906.

CHAPTER 2

Carryl, Frank M. How to Use a Light Microscope, Decatur, Georgia, By the Author, 134 Burton St., 30030, 1985.

Ford, Brian J. The Optical Microscope Manual: Past and Present Use and Techniques, London: David & Charles Ltd., and New York: Crain, Russak, & Co., Inc., 1973.

Gage, Simon Henry The Microscope, Ithica New York: Comstock, 1941.

Grave, Eric V. Discover the Invisible: A Naturalist's Guide to Using the Microscope, Englewood Cliffs, N.J.: Prentice-Hall, 1984.

Herman, Brian, and Jacobson, Ken, editors. Optical Microscopy for Biology, New York: John Wiley and Sons, Inc., 1989.

Hoffman, Robert. "The Modulation Contrast Microscope: Principles and Performance," Journal of the Microscope, Vol. 110, Pt. 3, (August 1977), pp. 205-222.

Hoffman, Robert, and Gross, Leo. "Reflected Light Modulation Contrast Microscope," Microstructural Science, Vol. 4, pp. 287-295.

James, J. Light Microscopic Techniques in Biology and Medicine, Amsterdam: Martinus Nijhoff Medical Division, 1976.

Scarff, Deborah. "Economical Procedure to Convert a Standard Light Microscope to the Hoffman Modulation Contrast System," Journal of Biological Photography, 50, no.4, (October 1982), pp 131-34.

CHAPTER 3

Chayen, J., and Bitensky, L. Practical Histochemistry, Chichester: 1991.

Clark, George, and Kastin, Fredrick. History of Staining, Baltimore: Williams and Wilkins, 1983.

Clark, George, ed. Staining Procedures, Baltimore: Williams and Wilkins, 1981.

El-Badry, Hamed M. Micromanipulators and Micromanipulation, New York: Academic Press, 1963.

Gordon, P.F., and Gregory, P. Organic Chemistry in Color, Berlin: Springer-Verlag, 1987.

Gray, Peter. Handbook of Basic Microtechnique, New York: McGraw-Hill, 1964.

Holum, Hohn R. Elements of General and Biological Chemistry, New York: John Wiley and Sons, 1979.

Horobin, Richard W. Understanding Histochemistry: Selection, Evaluation, and Design of Biological Stains, New York: John Wiley and Sons, 1988.

Jones, Ruth McClung. Basic Microscopic Techniques, Based on Guyer, Michael F., Animal Micrology, Chicago: University of Chicago Press, 1966.

Lee, Arthur Bolles. The Microscopist's Vade-Vecum: A Handbook of the Methods of Microscopic Anatomy, Philadelphia: P. Blakiston, Son and Co., 1890.

Lillie, Ralph D.. H. J. Conn's Biological Stains, Baltimore: Williams & Wilkins, 1969.

Ross, Michael H. and Reith, Edward J., Histology: A Text and Atlas, New York: Harper International Edition, 1985.

CHAPTER 4

Bergner, J., Gelbke, E., and Mehliss, W. Practical Photomicrography, London: The Focal Press, 1966.

Delly, John Gustav. Photography Through the Microscope, Rochester, N.Y.: Eastman Kodak Company, 1988.

Freeman, Michael. The 35mm Handbook: A Complete Course from Basic Techniques to Professional Applications, Philadelphia: Running Press, 1980.

Photography as a Tool, Revised Edition, Life Library of Photography, Alexandria, Virginia: Time Life Books, 1983.

Photomicrography: An introduction to Photomicrography with the Microscope, 14th Ed., Rochester, N.Y.: Eastman Kodak Co., 1944.

Shillaber, Charles Patten. Photomicrography in Theory and Practice, New York: John Wiley & Sons, Inc., 1944.

CHAPTER 5

Burgess, Jeremy; Marten, Michael; and Taylor, Rosemary. Microcosmos Cambridge, New York, New Rochelle, Melbourne, Sydney: Cambridge University Press, 1987.

Mason, Clyde Walter. Handbook of Chemical Microscopy, Vol. 1, New York: John Wiley & Sons, 1983.

CHAPTER 6

Bold, Harold C., and Wynne, Michael J. Introduction to the Algae: Structure and Reproduction, second edition, Englewood Cliffs, N.J.: Prentice-Hall, 1985.

Curtis, Helena. The Marvelous Animals: An Introduction to the Protozoa, Garden City, New York: The Natural History Press, 1968.

Grzimek, Dr.Dr.h.c. Bernhard. Grzimek's Animal Life Encyclopedia Volume 1: Lower Animals, New York: Van Nostrand Reinhold Co., 1972.

Hall, Richard P. Protozoa, The Simplest of All Animals, New York: Holt, Rinehart, and Winston, Inc., 1964.

Margulis, Lynn, and Schwartz, Karlene V. Five Kingdoms: An Illustrated Guide to the Phyla of Life on Earth, San Francisco: W. H. Freeman and Company, 1982.

Seeley, Harry W., Jr., and VjanDemark, Paul j. Microbes in Action: A Laboratory Manual of Microbiology, New York: W.H. Freeman and Co., Third Edition, 1981.

Sleigh, Michael. Protozoa and Other Protists, London: Edward Arnold: a division of Hodder & Stoughton, 1989.

Thorpe, Neal O. Cell Biology, New York: John Wiley and Sons, Inc., 1984.

Tortora, Gerard J., Funke, Berdell R., and Case, Christine L. Microbiology: An Introduction, Menlo Park, California: Benjamin/Cummings, 1986.

Wilson, Carl L., and Loomis, Walter E. Botany, New York: Holt, Rinehart, and Winston, 1967.

BOOKS OF GENERAL INTEREST

Bodanis, David. The Secret House: 24 Hours in the Strange and Unexpected World in which We Spend Our Nights and Days, New York: Simon and Schuster, 1986.

Curry, Alan; Grayson, Robin; and Hosey, Geoffrey. Under the Microscope, New York: Van Nostrand Reinhold, 1982.

Gould, James L., and Gould, Carol Grant, editors. Life at the Edge: Readings from Scientific American Magazine, New York: W. H. Freeman and Company, 1989.

Gray, Peter, ed. The Encyclopedia of Microscopy and Microtechnique, New York: Van Nostrand Reinhold Company, 1973.

Jírovec, O. Boucek, B., and Fiala, J., translated by Schierl, Margaret, Life Under the Microscope, London: Spring Books.

Stehli, Georg, translated by Vorderwinkler, William A. The Microscope and How to Use It, New York: Dover Publications, Inc., 1970.

BOOKS FOR YOUNG MICROSCOPISTS

Berman, William. How to Dissect: Exploring with Probe and Scalpel, New York: Prentice Hall, 1984.

Bleifeld, Maurice. Experimenting with a Microscope, New York: Franklin Watts, 1988.

de Kruif, Paul. Microbe Hunters, San Diego: Harcourt Brace Jovanovich, 1926.

Headstrom, Richard. Adventures with a Hand Lens, New York: Dover, 1976.

Headstrom, Richard. Adventures with a Microscope, New York: Dover, 1977.

Johnson, Gaylord, and Bleifeld, Maurice, revised by Beller, Joel. Hunting With the Microscope, New York: Prentice Hall, 1980.

Kelley, Alberta. Lenses, Spectacles, Eyeglasses, and Contacts: The Story of Vision Aids, Nashville: Thomas Nelson, Inc., 1978.

Rainwater, Clarence. Light and Color, New York: Golden Press, 1971.

Contents