v:; ..* ..* 9 :.@' :.@' . J .,: > ? -4 4 , 5 - t . ?i. t ! + ii,: r'I. i I f. Proceedings-of-me First Workshop/Conference of the Association for Biology Laboratory Education (ABLE) . .- Jon C. .Glase TESTED STUDIES FOR 1,ABORATORY I7F!ACHING Proceedings of the First Workshop/Conference of the Association for Biology Laboratory Education (ABLE) Edited by Jon C. Glase, Ph.D. Senior Lecturer and Laboratory Coordinator Section of Neurobiology and Behavior Division of Biological Sciences Cornell University KENDALL/HUNT PUBLISHING COMPANY Dubuque, Iowa, USA Toronto. Ontario, Canada Copyright (D 1980 by KendallIHunt Publishing Company Library of Congress Catalog Card Number: 80-82832 ISBN 0-8403-2271-2 All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without the prior written permission of the copyright owner. Printed in the United States of America We dedicate this volume to hardworking, effective laboratory biology teachers everywhere. Contents Foreword xi Preface xv L- Chapter 1 A Review of Sources of Living and Supplementary Materials for Laboratory Instruction with a Note about Vivisection 1 Don Zgelsrud Key Words: living organisms (sources),marine organisms (sources),sea urchins (sources),insects (sources),frogs (sources), animal organs (sourcesj, algae (sources), cycads (sources), dissection kits (sources),films (selected),film-loops (selected), vivisection (teaching). Reliable sources (based on 12 years teaching y experience) are given for protozoa, marine organisms, clams, sea urchins, nematodes, rotifers, earthworms, insects, crayfish, ammocoetes larvae, trout, frogs, rats, anesthesia, organs, algae, fungi, Physarum, plants, chromosomes, local resources, dissecting kits, films, film loops, charts and models. Materials for introductory I biology and zoology are stressed. Living materials are emphasized I and their ethical use is discussed. J 2 Use of Nitella in Biology Laboratory Classrooms 23 Charles E. Burr Key Words: IVtglLa, Cha~acid-base formation, OH- eflux. Hf t extrusion, protoplasmic streaming, cytoplasmic streaming, cyclosis. The freshwater genera Nitella and Chara are macroscopic algae which have giant, coenocytic internodal cells. Two striking physiological features of these cells are easily demonstrated: (a) the formation of external acid and base in discrete, alternating bands along the cell length, and (b) protoplasmic streaming. Directions for the preparation of these demonstrations are given, along with a method for culturing Nitella clavata. 3 Structural Organization of Living Cells 33 Osiris Boutros and Susan Noblit Boutros Key Words: cell organelles, living cells, structural organization of cell, polarizing light microscopy, phase contrast microscopy, cyto- chemistry, vital staining, photomicrography. Most of the unique vi Contents features of eukaryotic cells are visible in the light microscope. In addition to examining and identifying cell organelles and cell structures, students are introduced to the techniques and concepts of microscopy and cyto-chemistry through the use of phase and polarized light microscopy, vital stains, and reagents. Photomicrography and video cameras are discussed as valuable aids in teaching microscopy. 4 The Use of Plant and Animal Tissue Culture in Biology Laboratory Instruction 55 Lester C. Eddington Key Words: tissue culture, cell culture, in vitro culture, monolayers, cell propagation, plant cloning, phase microscopy, cytological staining, photomicroscopy. The theory and application of tissue culture as a method of teaching cellular concepts and methodologies are examined. Specific emphasis is given methodologies of animal tissue primary and subcultures, basic tissue culture equipment, cell and tissue staining, and microscope photography. 5 Use of a Plant Pathogen to Examine Koch's Postulates 89 Daniel D. Burke Key Words: Koch's postulates, pathogen, Erwinia, soft rot. This exercise describes the use of carrot as a model system in which to demonstrate Koch's postulates. The pathogenic organism used is Erwinia carotovora which causes soft rot of vegetable tissue. The ability of Escherichia coli to grow on carrot, but not rot the tissue, is also demonstrated. 6 Digestive Enzymes of the Cockroach 111 C. Leon Harris Key Words: digestion, enzymes, amylase, lipase, sucrase, protease, cockroach, statistics, sign test. Sites of enzyme secretion in the cockroach digestive system are determined by comparing enzyme activities in homogenates of salivary glands, crop, midgut and hindgut. Each group of students prepares crude enzyme extract from the four sections of the digestive system and tests for either sucrase, lipase or amylase and protease activity. Data giving the activity for each section relative to a control are compiled from all groups. The sign test is used to determine which section of the digestive system secretes each enzyme. Contents vii 7 Effects of Fasting and Exercise on the Depletion of the Liver and Muscle Glycogen in the Mouse (Mus musculus) 125 Bruce B. Virgo Key Words: glycogen, glucose, energy metabolism, fasting, exercise, liver, muscle. The purpose of this exercise is to determine the differential effects of feeding, fasting, and exercise on hepatic and muscle glycogen and to use these data to infer the biochemical mechanisms underlying the differences. The method is simple, reliable, and inexpensive. Tissue samples from fed, fasted, and exercised mice are weighed and digested in hot 30% KOH containing Na,SO,. The glycogen is precipitated from the digestate with ethanol and pelleted by centrifugation. The supernatant is discarded and the pellet dissolved in water. Aliquots of this solution are reacted for 10 min at 100' with anthrone in concentrated acid. The absorbance of the resulting blue complex is measured at 650 nm and compared to that of a standard glycogen solution. The concentration of glycogen in the tissues is calculated and compared. 8 A Rapid and Simple Procedure for Preparing qertebrate Karyotypes 137 Joseph R. Larsen and Carl Gilbert Key Words: vertebrates, karyotypes, chromosomes. A simple and rapid procedure for preparing vertebrate karyotypes is presented. The procedure involves arresting bone marrow cells in metaphase with a mitotic inhibitor, sacrificing the animal, recovery of the bone marrow cells, fixation, and staining with carbol-fuchsin. This procedure allows students to obtain karyotypes from vertebrates, study the species chromosome morphology, and estimate the species chromosome number in a single three-hour laboratory period. 9 The Elucidation of a Biochemical Pathway 145 Marcia K. Allen Key Words: gratia marcescens, biochemical pathway, prodigiosin. The elucidation of a biochemical pathway is performed using color mutants of Serratia marcescens. This technically simple experiment enables the students to order a series of mutant alleles in a biosynthetic pathway by a simple plating technique. Only basic sterile technique is needed. The principles of the one gene-one enzyme theory are demonstrated without the use of complicated biochemical extractions. This exercise could be performed anywhere where there are sterilization facilities. viii Contents 10 Brassica campestris L., a New Plant for Teaching Genetics 155 L.V. Crowder, J.A. Hawk, T.M. Gradziel, and S.E. Fast Key Words: introductory genetics, Brassjea mutants, techniques and procedures. An early-flowering (14-16 days) selection of Brassica campestris L. (field mustard or turnip rape) is suitable for completing inheritance studies during a 10-week quarter or a 15- week semester. Mutants are available for studying such principles and concepts as dominance and recessiveness, segregation and independent assortment, nuclear and extrachromosomal inheritance, linkage and epistasis, pleiotropic effects, penetrance and expressivity. The ease of culture, small plant size, flower characteristics, short life cycle of about 40 days, and prolific seed production are qualities that make this plant useful for students to design and carry out their own independent project, record observations, maintain records and interpret data. 1 1 Uses of the Honey Bee (Apis mellifera) in Teaching Introductory Biology Laboratory Courses 171 Jon C. Glase Key Words: honey bee (&is mellrferaj, observation colonies, investigative studies, honey bee morphology, electroantennogram. This chapter describes some uses of the honey bee (Apis mellifera) in the teaching of introductory biology laboratory courses and includes information on methods for establishing, maintaining, and using observation colonies within laboratory environments. In addition to specific suggestions for studies of phenomena occurring within observation colonies, a simplified form of the electroantennogram bioassay technique and its use in pheromone studies of the honey bee are described. 12 Dual Purpose (Process and Content) Laboratory Exercises: Animal Behavior Experiments Using the Hermit Crab, Pagurus longicarpus, the Land Snail, Polygyra sp., and the Isopod, Armidillidium vulgare 209 Eugene H. Kaplan Key Words: chi-square, null hypothesis, levels of confidence, habituation, anthropomorphism, ecological niche, limiting factor, herbivore, isopod, taxis, kinesis, T-maze, rotting log community. Two exercises are described. Each has two primary objectives: to teach an epistemological (process) aspect of science and to teach biological content. The first exercise presents the chi-square