SUBJECT: Mitosis and Meiosis Labs
DATE: 2/95; 2/97

I'm looking for help in finding a long-term solution to a perennial problem in our introductory labs. Our students, majors and nonmajors alike, THINK they know all about mitosis and meiosis, because they've heard the terminology in high school. When pressed for details, however, they're clueless. (The worst thing is that many of our graduate TA's are clueless, too, even though THEY think they've got it!) Even so, they uniformly despise any and all labs that attempt to cover this subject ("we know all this," "we did this in high school," etc.).

Here's what we've tried so far, in various combinations:
1. Prepared slides (root tip, whitefish blastula, lily life cycle), to identify and estimate the time spent in various stages.
2. Pop-bead chromosome models (available from Carolina Biological)
3. Having students make their own squash preparations from pretreated (with paradichlorobenzene) and unpretreated root tips, and Chlorophytum (spider plant) buds.

I figure we've got to develop a lab that teaches them the fundamentals of these processes without letting them know that WE know that they don't know it yet...:) If you've had success with a mitosis/meiosis lab, I'd LOVE to hear about it!

Thanks in advance,
Nora Ann

Nora Ann Bennett
Department of Biology
College of William and Mary
Williamsburg, VA 23187
(804) 221-2219
FAX: (804) 221-6483
EMAIL: nabenn@facstaff.wm.edu (as of 2/13)



Nora Ann Bennett asks about successful ways to teach mitosis and meiosis in labs. While the following doesn't directly show either of those processes, I've found students to get fairly excited (as excited as jaded intro biology students CAN become) when beginning such labs with a human chromosome spread. CellServ, in the Center for Advanced Training at Catholic Univ. of America, offers a kit with cultured HeLa cells. Students can make a stained spread and see many cells in interphase and stages of mitosis. HeLa cells are weird because their chromosome numbers are highly variable, but students are generally very interested in these tumor cells. This can also lead into discussion of controls of cell division. CellServ needs several weeks advance notice in order to produce the cells. Contact them at (202)319-5725, FAX (202)319-5721.
Address: CellServ
The Center for Advanced Training
103 McCort Ward Bldg.
Catholic Univ. Of America
Washington, D.C. 20064

The procedure for using this also appears in a Wadsworth Publishing Co. lab manual by Perry and Morton that accompanies the biol. textbooks by Starr and by Starr and Taggart.
Joy Perry
Univ. Of Wisconsin Center - Fox Valley
joyperry@uwcmail.uwc.edu
(414)832-2653


In reference to your problem with Mitosis and Meiosis, take a look at the Mac program titled Mitosis and Meiosis by Intellimation at PO Box 1530, 130 Cremona Dr. Santa Barbara CA 93116-1530. I have found using their program in class and lab has raised the learning curve for the processes in both major and nonmajor courses. I have been pleasently surprised by the apparent increase in understanding of Mitosis and Meiosis.

George C. Boone
Susquehanna U.
boone@einstein.susqu.edu


Nora Ann's note about Mitosis and Meiosis hit several hot buttons in me. The first question is, I guess, what do you mean by the "fundamentals" of these processes? Are you interested in the processes of chromosome movement, the segregation, crossing over, etc., or the functions that are carried out through these mechanisms.
More importantly, from my perspective, is my feeling that one should never mention Mitosis and Meiosis in the same sentence, or the same lecture. The coupling of the processes (because of their mechanical similarity) often confuses students about their very different functions. Mitosis is a process that is associated with cellular duplication, with equal distribution of chromosomal material to succeeding generations. Its primary function is in growth of the organism. Meiosis, on the other hand, is a rather different process whose function is to create haploid organisms from diploid, for the purpose of sexual reproduction.
It has always seemed to me that these processes should be presented in discussions that are appropriate to the function of each. i.e., discuss Mitosis when you are talking about growth, and save Meiosis for a discussion of reproduction. Then, it is possible to discuss how Meiosis is a varient of Mitosis -that some of the mechanisms i.e. use of a spindle, are similar, and that others, such as the chromosome alignment, are different.
Regardless of this quibble, I find that the most dramatic way to demonstrate the power and wonder of either process is to show some of the excellent time-lapse videos of mitosis. Among the best images are those from Jeremy Pickett-Heaps, but there are others. I would combine the videos with real life observations, such as sea urchin cleavage.

Joel B. Sheffield
Biology Department
Temple University
Philadelphia, PA 19122
jbs@sgibio.chem.temple.edu or
v5415e@TEMPLEVM.bitnet
(215) 204 8854



We have used mastery quizzes with yarn models of mitosis and meiosis to get some pretty spectacular group learning situations. At the beginning of lab, or at the previous lab meeting, students are given a list of quiz questions that involve the yarn models in demonstrating the answers. They are working in groups and they are told to prepare themselves to respond to any of the possible questions. When the group feels prepared, the individual who will do the demonstration (take the quiz) is selected by drawing straws and the question is drawn out of a hat (literally!).

Under these circumstances the questions can be pretty challenging. For example, "Use a yarn model to explain and illustrate why meiosis I is sometimes called the heterotypic division and meiosis II is sometimes called the homotypic division. Be prepared to explain how the term'reduction division' relates to these other terms." We didn't define any of the terms, but gave students access to a dictionary. Another question I liked was "Using a yarn model for an organism with 2N = 4, create two gametes with distinctly different chromosomes. Work the model through one generation, producing gametes unequal in numbers of grand-paternal and grand-maternal chromosomes."

Groups really sought out instructor help, which was freely given but only through aggressive questioning (What does this represent, What happens next?) and once they had worked out a solution, group members took turns quizzing each other on it, and often got into questioning each other rather aggressively also. It worked. We've done it with and without actually giving a grade for the final performance. The approach works just as well with questions related to microscopy preparations.

Robert B. Ketcham rketcham@strauss.udel.edu
Department of Biology (302) 831-2377
University of Delaware
Newark, DE 19716-2590


I have made a number of chromosomes from poster board that I have colored with different colors of magic markers to indicate dominate and recessive alleles. I give each set of students a diploid set and then tell them to prepare to undergo mitosis. It usually takes them a minute to realize that they need to replicate them first. I then distribute another set to represent the replicated chromosome (they are given tape to hold the sister chromatids togather, but velcro would be easier). I then have them sort the chromosomes at each stage and we move through the class to make sure they have the chromosomes lined up properly. At the completion of mitosis, we find out how many cells we end up with and what is their gentic constitution compared to the parent cells This seems to help students visualize the sorting process.

For meiosis we start over again with replication again (this time they catch on right away). We then tell them to arrange the chromosomes as they might appear at prophase I, metaphase I, anaphase I, etc. At this point we let them make mistakes in segregation (that is we do not correct their mistakes) so that we can illustrate the proper segregation of chromosomes. We then have them complete meiosis I and meiosis II and we then take a poll of the class to determine the genetic constitution of each of the daughter cells. If we have a large enough class, this allows us to nicely illustrate recombination and distribution of alleles. This is especially important for us because the next week we deal with genetics.
--
James M. Bader
Asst. Director, Center for Biological Education
Case Western Reserve University
jxb14@po.cwru.edu



I use post-it notes in my non-major's class to help demonstate WHY chromosomes pair during meiosis. Students label a set of post-it notes to represent a pair of alleles. Initially I have them scatter them randomly on a table top (up-side down). Working in pairs, each student picks up half of the notes. Even with just three sets of genes it is rare that a student gets one copy of each gene. I then have them stick the alleles together and repeat the selection. This time each student gets one note from each of the pairs. Of course, they both must get one of each allele using the paired notes.

I have also had students attempt to model the process of meiosis using post-it notes, but frankly I do not think that the steps of the meiosis are as important as the logic of the process.

Richard Weisenberg
Dept. of Biology
Temple University



I'm glad you brought up the subject of teaching mitosis and meiosis. we haven't solved the problem, but I second the idea of using time lapse films. We have several antique film loops of both processes and they really do help to get the idea across. I also put out sets of photographs of the different stages of meiosis which are labelled on the back. I tell the students to examine them and then to shuffle the photos and try to get them back in the correct order without looking at the labels. We also do a root tip squash and pipe cleaner chromosomes. But my favorite is the "Dance of the Chromosomes". I divide the students up into sets of eight and label them either A, a, B, or b. Then I tell them to go outside and pretend to be chromosomes going through meiosis. I let them figure out what to do with the minimum of direction and then have each group perform for me. Most of them find this silly but fun, and even the ones who feel it's rather an affront to their dignity have grudgingly admitted that it did help them to visualize the process. I certainly enjoy it. Good luck and please pass on any ideas you find work.

Susan Schenk
Claremont Colleges
Claremon, Ca
sschenk@jsd.claremont.edu


We do the 'dance of the chromosomes as well here at Bates. Yes, its silly but they really do enjoy it. We prep them for this dance by having them watch a video of a humerous dance/skit filmed by a senior rhetoric student a few years back......she needed a science credit to graduate and we had her produce the mitosis/meiosis skit as part of her effort. After all that work...she REALLY does understand the two processes.
Joe P
--
Joseph G. Pelliccia PP-SEL-IA * "A beginning is a delicate time"
Chairman, Department of Biology * -Princess Irulan
Bates College, Lewiston, ME 04240 * DUNE




I've been following the Mitosis and Meiosis thread and my understanding of Nora's problem is not so much How to teach this material but rather, How to keep students who have an inflated sense of their understanding open to further exploration?

I have had good success with using cancer biology as a hook to get them into cell cycle/cell division material without ever saying the m-words. Sometimes I come at it from forensic toxicology. What would you expect to find in tissue killed by colchicine? I like to hook them with nice-to-know mind-candy and then I build in the need-to-know biology. (Of course I'm clear with them about which is which.)

In the lab, I think it is hard to beat some type of investigative exercise in these situations.

How about giving someone a handful of beans or a spider plant (or prepared slides), a microscope etc. and expect them to ask a question for which they can find an answer with the tools provided. Yarn models, pop beads, video, modelling clay, and chromosome dancing etc. may all be used for background or to help make predictions. The theory goes that if students can be sold on asking their own questions and then doing their own science they will admit/discover their misconceptions and work to straighten themselves out. This has been my experience with a large Year 2 genetics class.

We have done investigative style labs with the Vicia root tips/colchicine system. Students investigated questions relating to the proportion of cells in various stages, chromosome condensation, mitotic index variation in different root tip sections as well as effects and kinetics of colchicine treatment. There are also estimates of chromosome number and karyotypes etc. to be done.

I would like to expand our repetoire in this area. I'd appreciate further information on the spider plant bud exercise as well as leads on other cytogenetic systems that would lend themselves to student questioning. (Preferably not involving scraping marrow out of the broken bones of furry animals).

Tom Haffie Phone: 519-679-2111 (6502)
Department of Plant Sciences FAX: 661-3935
University of Western Ontario e-mail: thaffie@julian.uwo.ca
London, ON, Canada
N6A 5B7


Tom,

For Chlorophytum, 2N = 28.

Nora Ann

Nora Ann Bennett
Department of Bioloyg
College of William and Mary
Williamsburg, VA 23187
(804) 221-2219
email: nabenn@facstaff.wm.edu


I agree with Joel Sheffield that the key to making mitosis/meiosis labs work is knowing what it is you are really trying to stress. I disagree with the idea that mitosis and meiosis shouldn't be mentioned together, precisely because they are so easily confused- I think the best way to teach students to discriminate between two similar things is to put them side by side and carefully point out the differences.

In contrast to Nora Ann we've had success with the Carolina popit beads (we used to use pipe cleaners to simulate chromosomes but the beads are better for showing crossing over). I have the students start by making a model of a G1 phase nucleus with single chromosomes as a reference. They then work together to make models of the various phases of mitosis ending with 2 daughter nuclei. They should then see that the daughter nuclei are identical to the reference nucleus. They then repeat the process with meiosis and find that this time they end up with 4 haploid nuclei that are all different from the reference and (thanks to crossing over) different from each other. The time and effort they invest in making all the models lined up together on the bench top and the graphic differences in outcome seem pretty effective in helping them remember which process is which and what each process is good for.

I haven't encountered too much resistance from the students. They're often so confused after the lecture hall presentation on M&M that they are happy to have a hands on opportunity to straighten it out. TAs are another matter. In the prelab meeting I tell them that even though they may have mastered this as freshmen, that was years ago and they probably haven't used it much since then. I urge them to review before they get in front of their classes rather than let themselves look stupid by getting confused at the last moment. Since I can tell them that this has happened in the past several times and that the TAs involved really did feel pretty embarrassed, they generally heed the warning.

John Dickerman
Biology Lab Coordinator
Northern Illinois University
T80JWD1@WPO.NIU.EDU
(815) 753-3101



I'm looking to have students design their own study of cell division (thinking of next Fall). How much time do you allocate? I'm thinking of 2 to 3 weeks (1 3hr lab per week), to introduce the technique of staining and squashing a root tip, time for students to confront the decisions necessary for testing the effects of a chemical, and a full lab to execute.

Do you send your students out to do some research into methods - chemicals to use, concentrations, length of exposure, etc.? Are all your studies looking at the effects of a chemical?

Do you let them develop their own methods of quantification? Do you always use squashes or are you doing sectioning? Squashes are so easy and they are fine for catching cells in interesting poses, but students get befuddled when asked to do counts/area or percentages - the sampling question of what is a suitable area is a stumper. The crisp geometry of thin section makes this easier but too much is lost in the transition from squash to commercially prepared slides - the latter are no longer the students' project.

Related to the quantification, one of the appeals of doing this experimentally is the opportunity to address bias in sampling. Students can do a 'blind' study, which to me is the solution for the sampling problem. But I've found students don't discover this on their own. What do you do?

We've been using onion sets, which are great. We can give a student a set in a bit of vermiculite in a cheap plastic drinking cup and tell them to moisten vermiculite thoroughly 2-3 days before lab and Bingo! students have prepared their own material. [Actually we have yet to do this with the students. We did do it with the instructors and its so easy it would work with students (next year). This year I encouraged instructors to have their students use this material in experiments but none did - they used it for things like quizzing students on microscopy.] Do you have students sprout their own beans?

As I understand it, seedling roots of many species will work. Grocery store sprouts don't, unfortunately - they aren't growing any more. I'd be interested in hearing about materials that others use.

RKetcham/UDelaware



At the risk of this thread turning into a knot, I'll respond to Bob
Ketcham's posting about M and M labs;
>
We've had pretty good luck with introducing the I-lab as creative tool-using. Our standard format is to make a big deal of the central role of questioning in research and the nearly complete absence of questioning in science education. We are very direct with students that I-labs require the use of unfamiliar tools as well as familiar tools in unfamiliar ways. We set them the task of asking their own questions and then assure them that much help is available.
We meet for 2 hr early in the week and then again for 3 hr later in the week.
In lab #1, we open with some community building among students and instructors and then set the general biological stage - the conceptual tools - videos, brainstorming, concept maps etc. Then, just as people get restless, we break out the technical tools and roll up sleeves for the rest of the (first)lab.

Lab #2 is devoted entirely to a "dry run" of the techniques for practice - similar to a standard "cook book" lab. This exercise may also provide data or material to be used in future labs. During down times in the protocol, we mingle and discuss how these techniques might be applied to their own questions.

Before lab #3, many students worry and fret about their research question but at least they arrive in an attentive frame of mind. We then spend some time on general experimental design with some specific (obvious) examples that pertain to the system being studied in the lab. In this "design lab", I have a 36 students and 3 - 5 well briefed instructors. It is an intensive, hand to hand, one on one, small group, teaching environment that leaves everyone exhausted. It's great! In reality, there are only about 6 or 8 appropriate questions and the 36 individual students coalesce into subgroups based on common interest.

Lab #4 and #5 is devoted to confirmation of designs, experimentation and, perhaps, for extensions or repeats of the original designs. Students collaborate for the technical work if they like. Groups of 1 to about 5 depending on the design and personalities.

Lab#6 is time for making sense of the data and marshalling the persuasion tools to be used in Lab #7 to convey the science to instructors and peers in a Symposium. We use such instruments as oral presentations, poster sessions, and personal interviews as well as some some of writing.

This probably sounds like a luxuriant amount of time for one exercise but we are also working specifically on scientific writing and a GCK exercise is playing out in the background. Students then move from my genetics area into a 4 week session in ecology and then another session in cell biology.

>Do you send your students out to do some research into methods -
>chemicals to use, concentrations, length of exposure, etc.? Are all your
>studies looking at the effects of a chemical?

We do our best to assume only background covered in prerequisite Year 1
biology courses. Specialized information or data that might be expected to
arise from a preliminary experiment is often provided or referenced as
appropriate.

Chemicals are just one of many variables that we use to broaden the range of good questions that can be asked.
>
>Do you let them develop their own methods of quantification?

We would present established methods as a "tool".
Do you
>always use squashes or are you doing sectioning?
Squashes only. We satisfy ourselves with very crude scale (1 mm +) sections for sampling.

I am mostly concerned with the thinking that goes on in the lab. If a student can ask a good question and find the answer by analysing prepared slides then I'm happy. They have done the technical practice session - I can let go of the rest.
>
I lead them into discovering such things on their own with a couple of well aimed criticisms of their designs. I point out how they will be vulnerable when it comes time to present the data to peers.

>Do you have students sprout their own beans?

We didn't, but we will. I'm anticipating the look on their faces as their initial confusion clears as they figure out that they can get cycling cells by germinating the seeds.

Sets sound great. How many chromosomes? Big ones?

>As I understand it, seedling roots of many species will work.

We are having some difficulty finding shoots with few, fat chromosomes.
Vicia faba is wonderful, Corn is OK, Some types of peas are OK. Commercial
crop species tend to be polyploid.

Tom Haffie Phone: 519-679-2111 (6502)
Department of Plant Sciences FAX: 661-3935
University of Western Ontario e-mail: thaffie@julian.uwo.ca
London, ON, Canada
N6A 5B7



Tom, thanks for the detailed reply. I really like what you are doing and would love to dig even deeper into specifics but perhaps that should be done as a private chat at a future time.

I would like to pick up on one thing you say and turn it into a general discussion if others can offer insights. It has to do with having students work in groups. As I read Tom's description, his students are talking, thinking, questioning, and practicing techniques for the first 3 labs, and this involves small group interactions, probably of shifitng compositions. When they lock in on their own investigations, in labs 4 and 5, they work collaboratively at their own discretion ("Students collaborate for the technical work if they like. Groups of 1 to about 5 depending on the design and personalities.")

I would like to have this happening in my labs, but I'm not there yet. We've used groups for several years, but have been in a more structured mode. We assign students to a group (5 per), have each group write a 'group contract' in which people define the level of commitment they are willing to make to the group effort of designing and conducting an investigation. This is not bad, and I will probably continue to start the course this way (freshman, by the way), but I would like to move on to a more voluntary arrangement of groups. What I'm guessing has to happen is that the whole lab section needs to be functioning as a group at the beginning - working in one area, exploring what they do and do not know already, exploring possibilities, and drafting potential experiments - the give and take, trial and error preludes to making decisions. Then they can break out into voluntary groups to do different things, but they will still be able to talk and consult across groups, and will be able to communicate their results to an informed audience. As I say, I'm just trying to work these things out for my situation. Tom Haffie's comments gave me a boost of confidence (thanks again, Tom). I wonder if others agree or disagree with the reasoning.

Tom, you did ask a specific question about the number of chromosomes in the onion. As my daughter would say, DUHHHHH! I think its in the twenties (26?) - that would fit with what I saw, probably countable but not easily. Maybe someone else has the number at hand?

rketcham, Univ. Delaware

According to the Chromosome Atlas of Flowering Plants, most
species of onion (Allium) have 2N=16, but the number varies
from 14 to 48. I knew this book would come in handy!!

--
James M. Bader
Asst. Director, Center for Biological Education
Case Western Reserve University
jxb14@po.cwru.edu

Several years ago, we decided to have our non-majors make their own slides
to observe mitotic figures rather than looking at prepared slides. We found that onion roots worked pretty well, but that narcissus roots worked even better. The cells in narcissus roots are larger with correspondingly larger chromosomes than onions. Almost everyone in class got a nice prep with visible figures.

Narcissus bulbs should be prepared 48-72 hours before lab. To stimulate root growth, lightly scrape off (with a scalpel) the roughly circular callous found within the circle of old roots. Then place the bulb in water. In a warm room, roots should begin to grow after about 24-36 hrs and be usable in 48. After the roots have reached about 4 cm or so, mitotic figures are difficult to locate The only problem with using narcissus roots is that the bulbs are not available year round. However, they will keep in the refrigerator, so you can purchase them ahead of time.

Not only do narcissus roots have larger chromosomes than onions, but in another week or so, you have beautiful flowers!

Doreen Schroeder
Department of Biology
Mail # 4327, University of St. Thomas
2115 Summit Ave.
St. Paul MN 55105




I've really enjoyed the discussion on M & M labs, particularly the investigative labs done by Tom Haffie and planned by Bob Ketcham. Their discussion has inspired me to bring mitosis back into the laboratory. We used to do one lab on cell division and the cell cycle using onion bulblets purchased at garden stores or home improvement stores (just wrapped them in moist paper towels for 1-2 days). Students made their own squashes, and calculated mitotic indeces and the duration of each phase (based on a 16-hour cell cycle for onion). However, it was at this time that one of the lecturers in our team-taught course kept on trying to squeeze mitosis out of the lecture entirely, suggesting that it be covered solely in a demonstrative lab complete with an intro lecture by me. Being that I was in a contrary mood at the time, I argued that a non-investigative lab was what I was trying to get away from, and that mitosis should be taught in lecture, using some of the techniques (pop-it beads, yarn, dances, shuffled pictures) that have been discussed here. This would provide a chance to do some group work and active learning in lecture -- a great break from the "normal" lecture routine. I ended up dropping mitosis entirely from the lab last year -- to create more space for 2-3 week investigative labs. Now, Tom's and Bob's ideas will push me to try and bring back mitosis and cell cycle next year as a several week investigation. Thank you!

(Incidentally, we teach mitosis & meiosis separately. Meiosis is covered in our "organismal" fall course when Mendelian genetics is taught, and mitosis is taught in our "cell/molecular" spring course when we discuss the cell cycle.)

It seems we constantly battle with trying to blend (at least!) 2 goals of the teaching laboratory: allowing students to more fully comprehend concepts presented in lecture by hands-on experience with those concepts after lecture; and allowing students to experience and learn the critical thinking skills used in science by designing, performing, and analyzing their own experiments, often with no regard for what's being covered that week in lecture. I find it very hard to have a few "investigative" labs in a row, and then go back to a "concept-reinforcing" lab -- the students often object. On the other hand, if I have a few concept-reinforcing labs in the beginning, and then switch to investigative labs, students argue that they need and rely on the laboratory to help them with the lecture material. I tend to lean towards (if not exclusively) the investigative lab, but then feel guilty that I'm not doing all I can to help students with as many concepts as possible from lecture. Do many people try a mix of the 2 types of labs? And if so, how do students respond? If you use all investigative labs, do students complain that "lab has nothing to do with lecture" much of the time?

Mike O'Donnell
Dept. of Biology
Trinity College
Hartford, CT 06106-3100
michael.odonnell@mail.trincoll.edu



Mike O'Donnell laments that his students complain that the investigative labs have nothing to do with the lecture. Maybe the problem is with the lecture!

Tom Haffie
thaffie@julian.uwo.ca



Thank you all for the wonderful discussion of mitosis and meiosis!

I heartily agree that mitosis and meiosis are too often lumped together, leading to massive confusion. In fact, I've argued that we should do as Mike O'Donnell suggested (our cell/molecular course is in the fall, and the genetics/ecology/evolution in the spring):

> (Incidentally, we teach mitosis & meiosis separately. Meiosis is covered
> in our "organismal" fall course when Mendelian genetics is taught, and
> mitosis is taught in our "cell/molecular" spring course when we discuss the
> cell cycle.)

Currently we do a semester-long inter- and intraspecific competition experiment with Drosophila population cages. One cage has two species of Drosophila, and the other has two morphs of the same species (red vs. white-eyed melanogaster). The students sample the cages weekly, then graph and analyze the data at the end of the semester. The weakness of this lab is that each lab section has one set of cages, and a different group of students samples each week. They don't really see any continuity, and they don't "get it" until the end of the semester when we pool the data for the semester. With over 400 students, though, we can't afford a cage for each group of 4-
6 students.

Tom asked about the spider plant bud exercise. This procedure was written by Stan Hoegerman, our cytogeneticist. Collecting the buds is the tedious part. We've got about a dozen plants in the greenhouse, and we collect the buds sporadically. We pick them with tweezers, and drop them into a fixative of 3 parts ethanol to 1 part glacial acetic acid. The fixative should be changed daily until it's clear (the chlorophyll, etc. will leach out). Once you've got clear fixative, you can store the buds indefinitely (from one year to the next).

In lab the students select a flower bud, place it on a microscope slide in a drop of 45% acetic acid, and dissect out the anthers (with freshmen we've got to be very careful to show them what they're looking for, since they've never had a botany class...). Once they've isolated the anthers, blot the acetic acid, and add a drop of aceto-orcein stain. Macerate the tissue with dissecting needles until you've got a fine pulp. Add a cover slip. Most of the cells will be in the same phase of meiosis, but different students will have different phases.

Our goal for this lab is much the same as when we have the students do their own root tip squashes: that they see where this process actually occurs, and that they learn some cytogenetics techniques.

I've enjoyed this discussion, and I thank you all for your generous comments.

Nora Ann

Nora Ann Bennett
Department of Biology
College of William and Mary
Williamsburg, VA 23187
(8040 221-2219
email: nabenn@facstaff.wm.edu


This should have been sent long ago when the topic on the table
was cell division, but for what it's worth, here it is.

I didn't see any mention of the very useful technique developed
by Joe Nickolas, Northland Community College, Holbrook, AZ, for
staining root tip squashes from Zebrina spp.(Zebra plant, Wandering
Jew, etc.). We have had great luck using this technique and we
especially like the fact that the students start with living
root tips and have a stained squash in about ten minutes.
The first-time success rate is high, but the short preparation time means
that "do-overs" don't cause unreasonable delays in the work if
students don't get good preps the first time. The preps
are pretty durable, too. I tried sealing the cover glasses
on several student slides and observing them after storage.
There were still decent, visible mitotic figures after several
weeks.

Now, the question. Was this technique, which was originally
presented at an ABLE mtg., ever included in an ABLE volume?
If it wasn't, maybe Joe could be imposed upon to put it on
this bulletin board. I think that many people would like it as a
replacement for some of the messy, smelly techniques that
employ pretty nasty fixatives or preservatives, or both.

Leland Johnson
johnson@inst.augie.edu

 

2/97: plant chromosome squashes
 
 
I seek suggestions on obtaining meiotic chromosome squashes from anthers of
spring flowering trees and shrubs.
 
Last spring two students and myself collected blossom buds in February, March
and April, fixed them in FAA, stained in iron acetocarmine, and squashed the
anthers. We had poor results.
 
David Crabtree
Hillsdale College
Hillsdale, Michigan
 
 
 
David,
 
Do you mean by "poor results" that your staining failed to show
chromosomes, or the cells were not in meiosis. Several years ago, an
ABLE meeting had a workshop on using Allium for meiosis labs, and
included techniques on growing the plants, fixing the flowers
and staining anthers. My own experience is that meiosis usually
occurs very early in the morning when the flower buds are barely
noticeable.
 
Hope this helps, Graham
 
----- "Graham R. Kent" <gkent@science.smith.edu




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