Invertebrate Anatomy OnLine
Supplies and Recipes ©
2jul2006
Copyright 2001 by
Richard Fox
Lander University  

Preface

            This is a chapter from Invertebrate Anatomy OnLine , an Internet laboratory manual for courses in Invertebrate Zoology.  Additional exercises can be accessed by clicking on the links on the left.  Terminology and phylogeny used in these exercises correspond to usage in the textbook by Ruppert, Fox, and Barnes (2004). 

Sources

BioQuip Products, 17803 LaSalle Ave, Gardena, CA 90248-3602 www.bioquip.com bioquip@aol.com 310.324.0620

Specializing in equipment for entomologists.

Carolina Biological Supply Co., tel = 800.334.5551, fax = 800.222.7112, www.carolina.com, 

Dissecting tools, living and preserved specimens, slides.

Charles D. Sullivan Inc. 6685 Holt Road, Nashville, TN 37211, tel = 615.832.0958. Living amphibians.

Fine Science Tools, tel = 1.800.521.2109, fax = 1.800.523.2109, info@finescience.com, www.finescience.com

Stainless steel minuten nadeln, stainless steel insect pins, dissecting tools.

Fisher Scientific, www.fishersci.com 800.766.7000

General Scientific equipment and supplies.

Fluker's Cricket Farm, P.O. Box 378, Baton Rouge, Louisiana 70821 tel = 800.735.8537

Living crickets.

Gulf of Maine, Inc., PO Box 321, Pembroke, ME.  www.gulfofme.com 207.726.4620

Living marine specimens. 

Gulf Specimen Marine Laboratories, Inc.  POBox 237, Panacea, Fl 32346, tel = 850.984.5297, fax = 850.984.5233, gspecimen@sprintmail.com, www.gulfspecimen.org

Living marine specimens. 

Leeches USA, 300 Shames Drive, Westbury, N.Y., 11590, tel = 800.645.3569.  

Living medicinal leeches.

Maine Bait Co., Box 204, Newcastle, ME 04553, tel = 207.563.3000, fax = 207.563.3055.

Living Glycera and Nereis for the bait trade.  Glycera (bloodworms) are $0.20 each, Nereis (sandworms) are $0.15 each.  No minimum order.  Send order attention Tom Dinsmore. 

Marine Biological Laboratory, Department of Marine resources, Woods Hole, MA 02543, tel = 508.289.7375, fax = 508.548.7371, www.mbl.edu/animals/index.html  go to Index of Services, ARD Catalog.

Living marine specimens.       

Nasco Science, 901 Janesville Ave., Fort Atkinson, WI 53538-0901 1.800.558.9595 or 1.920.563.8296 (fax).

Live and preserved materials, lab equipment, chemicals, microscopes, etc.

Nebraska Scientific, 3823 Leavenworth St., Omaha, NB 68105, tel = 800.228.7117.

Preserved specimens.

Orbis Scientific, Inc.  P.O. Box 3890, Milton, FL, tel = 904.626.0708, fax = 904.626.4634.   Provides Triops eggs, Artemia eggs, Daphnia eggs.

Pacific Biomarine, P.O.Box 1348, Venice, CA 90294, tel = 213.677.1056.

Living marine invertebrates. 

Santa Barbara Marine Biologicals, www.sbmarinebio.com Located in California this new company currently offers living specimens of Muricea californica, Strongylocentrotus franciscana, S. purpuratus, Lytechinus variegatus, Pycnopodia helianthoides, and Aplysia californica. The web site was being developed in March 2005.

Sea Life Supply, 740 Tioga Ave., Sand City, CA 93955, tel = 408.899.3399.

Living marine animals.

Selph's Cricket Ranch, Inc.  420 S. Front St., P.O.Box 2123, Memphis, TN 38101-2123, tel = 800.238.7322

crickets (Acheta domestica).

Sigma Chemical Co., P.O.Box 14508, St. Louis, Mo. 63178.

Biochemicals, including amaranth. 

Triarch, POBox 98, Ripon, WI 54971-0098, tel = 800.848.0810, fax = 888.848.0810, foreign fax = 920.748.3034

Microscope slides.

VWR Scientific Products, tel = 800.932.5000, www.vwr.com,

Scientific equipment, applicator sticks. 

Ward's Natural Science, POBox 92912, Rochester, NY 14692-8439, tel = 800.962.2660, fax = 800.635.8439, www.wardsci.com,

Dissecting pan wax, dissecting tools, living and preserved specimens, prepared slides. 

Recipes

            ACETIC ACID, 1%.  Mix 1ml glacial acetic acid with 99ml water.

            ACID FUCHSIN, ACIDIFIED 0.25%.  Mix 0.25g acid fuchsin, 1 ml conc. hydrochloric acid, and 100 ml distilled water.

            AMARANTH, 0.1% IN CRICKET SALINE.  Add 0.1g amaranth powder to 100 ml cricket saline solution.  Amaranth is available from Sigma.

            BALAMUTH'S pH 7 BUFFER.  For frog rectal contents, especially opalinids.  To 1500ml water add 7g sodium chloride (NaCl), 0.4g potassium chloride (KCl), 0.4g magnesium sulfate (MgSO 4•7H 2O), 0.1g potassium dihydrogen phosphate (KH 2PO 4), 0.1g sodium bicarbonate, and 0.2g calcium chloride (Wessenberg, H.  1961.  Studies on the life cycle and morphogenesis of Opalina.  Univ. California Publ. Zool. 61:315-370.)

        BRINE For Artemia cultures.  40 g non-iodized salt per liter of chlorine-free fresh water.  Use pond water, spring water, lake water, or aged tap water for this purpose.  Tap water should be allowed to sit for at least a week (in a container that provides a large surface area) before use.  

            CARMINE/SEAWATER SUSPENSION, 0.1%.  Mix well 1g powdered carmine powder with 1000ml seawater.  Shake well before each use.

            CHLOROFORM-SATURATED WATER.  An anesthetic for aquatic organisms.  Place 5 ml chloroform in 1000 ml water (seawater, pondwater, or tapwater, as appropriate) and shake vigorously.  The chloroform will sink below the water after shaking.  Use the water, not the chloroform, as needed. 

            CHLORETONE, 0.2%.  Earthworm anesthetic.  Dissolve 2g chloretone in 1000 ml pondwater.  Chloretone is 1, 1, 1-trichloro-2-methyl-2-propanol, or chlorobutanol, and is available from Ward's Natural Science Establishment.

            CHLORETONE, 5%.  Anesthetic for freshwater metazoans.  Dissolve 50g chloretone in 1000 ml water.

            ETHANOL, 5%.  An anesthetic for freshwater or marine invertebrates.  Add 53ml 95% nondenatured ethyl alcohol to 957ml pondwater or seawater.

            IODINE-POTASSIUM IODIDE .  Mix 6g potassium iodide, 4g iodine in 100 ml water.  Deep blue-black is positive test for starch.  Flagella and cilia stain with this solution.

            JANUS GREEN-NEUTRAL RED.  Mitochondrial stain.  1g neutral red, 0.5g Janus green, 100ml 95% ethanol.  Add stain to slide, allow to air dry.  Add culture to dry stain on slide.

            MAGNESIUM CHLORIDE, ISOTONIC WITH STANDARD SEAWATER.  Dissolve 77g MgCL 2•6H 2O in 1000ml distilled water or chlorine-free tapwater.

            METHYL CELLULOSE, 10%.  To immobilize protozoa.  To 10g methyl cellulose add 45 ml boiling water.  Let stand 30 min at 10 ° C., add 45 ml cold water and stir until smooth.  Bubbles will eventually disappear.

            METHYLENE BLUE, 0.5%.  Add 0.5 g methylene blue to 100ml water.  For nuclei and other organelles.

            METHYL GREEN, ACIDIFIED 1%.  Excellent nuclear stain but kills cells.  1g methyl green, 1 ml 1% acetic acid, 100ml water.  Methyl green without acid is a vital stain and does not kill cells.

            NEUTRAL RED, 1%.  1g neutral red in 100ml water.

            NIGROSIN, 10%.  For flagellate and ciliate pellicle surface pattern.  10g nigrosin, 100g water.  Add to preparation on slide and allow to air dry.  Observe dry without coverslip.

            PHLOROGLUCINOL-HYDROCHLORIC ACID.  For lignin, as a test for the presence of wood.  Prepare a saturated solution of phloroglucinol (1,3,5 trihydroxybenzene) in 20% hydrochloric acid.  Place a drop or two on the slide, mix and add a coverslip.  Lignin stains red-violet.

            PHOSPHATE BUFFER, 0.01M, pH 6.8.  Add 4.58g potassium phosphate and 4.67g sodium dihydrogen phosphate (NaH 2PO 4) to 1000ml water.  This makes a 0.5M stock solution.  Dilute one part of stock with 4 parts of water to make a 0.01M working solution.

            SALINE, EARTHWORM 0.70%.  Add 7.0g sodium chloride to 1000ml water.

            SALINE, BLUE CRAB. Callinectes Ringer's Solution:   2.444g NaCl, 0.086g KCl, 0.142g CaCl 2, 0.166g MgCl 2•6H 2O per 100 ml water at pH 7.3-7.5 (Pyle & Cronin, 1950).

            SALINE, 0.8-0.9% HOMIOTHERM.  Add 8-9g, as desired, sodium chloride to 1000ml water.

            SALINE, INSECT.  Dissolve 9g sodium chloride (NaCl), 0.2g potassium chloride (KCl), 0.27g calcium chloride (CaCl 2•2H 2O)  and 4g glucose.  Adjust pH to 7.2 with sodium bicarbonate (NaHCO 3) Add enough distilled water to bring final volume to 1000ml.

            SALINE, LEECH ISOTONIC (Hedon-Fleig's solution).  To 100ml distilled water add 7g sodium chloride (NaCl), 0.3g potassium chloride (KCl), 0.1g calcium chloride CaCl 2), 1.5g sodium bicarbonate (NaHCO 3), and 0.3g magnesium sulfate MgSO 4.

            SALINE, 0.5-0.7% POIKILOTHERM.  Add 5-7g, as desired, sodium chloride to 1000ml water.

            SALINE, WOODRING'S CRICKET.  A physiological saline solution for crickets.  Dissolve 9.1g sodium chloride (NaCl), 0.52g potassium chloride (KCl), 1.2g calcium chloride (CaCl 2•2H 2O), 0.82g magnesium chloride (MgCL 2•6H 2O) in 1000ml distilled water.

            YEAST/CONGO RED SUSPENSION.  Paramecium and rotifer food for visualizing feeding.  Boil 10g dry yeast with 0.1g Congo red in 30 ml water for 10 min.  Cool and store refrigerated until needed.  The suspension can be kept frozen indefinitely.

            YEAST SUSPENSION Daily Artemia food for maintaining cultures.  1 package baker's yeast suspended in 100 ml fresh water.