Click on the links above to go the the different labs for the Development Course. The format is a little funky, but you should be able to see what is being done in a given week. Below is an introduction to this semesters labs:

Developmental Biology Laboratory, Spring 2011 

Date                                    Weekly Topics                                                                   Page                                                                             

Aug 25             Dictyostelium - Cell Signaling                                                             5-12  

Sept 1              Drosophila - Heat Shock Proteins Expression                                   13-21 

Sept 8              Drosophila Imaginal Disk and Ecdysone                                           13-21 

Sept 15            Early Zebrafish Development/Protein Fingerprinting I                       22-33 

Sept 22            Protein Fingerprinting II                                                                       22-33 

Sept 29            Early Xenopus Development                                                             34-40 

Oct 6               Microscopic Examination Fertilization and Cleavage                        42-47 

Oct 13             Fertilization - Sea Urchin                                                                    42-47 

Oct 20             Microscopic Examination of Heart Development                              48 

Oct 27             In Vitro Chick Heart Development                                                     49-54 

Nov 3              Cell Surface Glycoproteins and Sponge Reaggeration                     55-61 

Nov 10            Presentations I 

Nov 17            Presentations II                                                                                   -- 

Dec 1              Buffer Lab

                                         Laboratory Notebooks for the Experimental Labs  

All students are expected to keep an accurate record of their observations and experiments during the experimental laboratories. The notebook need not be elaborate, but it should take into account the following important points.  

1.         The most important components of the notebook are things like dates, times, numbers and on-the-spot observations. You will find like most of us that memories provide a singularly bad scientific record.  

2.         Diagrams are very helpful in recording observations; they often save time and writing space. They need not be artistic, often a simple one serves quite well. Be sure to label things - the parts of your drawings and also the different dishes or test tubes that you use in the experiment. Make your drawings large enough to show all the details you can see.  

3.         After the results are recorded sit down and think about them. What conclusions can you draw from them? Write a coherent discussion of your results using correct grammar and spelling. Include a discussion of any "bad results", that is data contrary to that obtained by other students in the lab. This is important because bad results often provide useful information and insights.  

The laboratory notebook will be collected and graded and will be used as the basis of your weekly lab reports that explain the results of your labs. If you cannot express your ideas in the written word, how can they be useful to other investigators who want of continue where you left off? Be organized!

Often the lab exercises will ask you to perform many tasks. There is no time for you to sit in class and read the lab exercises for the first time. You will be expected to have a general flow chart that lists the key activities of that day’s lab. Include what kind of data or observations you are to collect before you come to the lab. Planned experiments work better and more efficiently. Most of all enjoy the lab. This is good stuff! Share your results and your excitement!  If you see something really neat show your neighbor or instructor.  

GENERAL LAB PROCEDURES FOR MOLECULAR LABS

Reagent Handling 

--          Wear gloves when handling all common reagents. 

--          Restriction enzymes are kept in alphabetical order in a "restriction enzyme" box.  Remove enzymes only when ready to use and place the tube in firmly packed ice.  Replace immediately.  Modifying enzymes are kept in the Stratacooler; when using these enzymes remove the Stratacooler when the enzyme is needed and immediately replace the lid and return to the freezer.  If the Stratacooler becomes covered with frost, it needs to be defrosted to prevent moisture from accumulating around the lips of the tubes.  

--          When attempting to hasten the thawing of a frozen reagent, rub the bottom of the tube between one's fingers; do not roll the entire tube between one's palms as this may contaminate the lip of the tube. 

--          Make a habit out of glancing at the pipette tip prior to inserting it into the enzyme tube to be sure that it is visibly clean.  

--          Frequently check and periodically clean the barrels of the pipette.  They should contain no moisture or dirt. 

--          Never place a pipette man with a used tip (or a "green thing" with a used glass pipette) in a horizontal position; fluid may leak into the barrel.  

--          If a particular reagent/enzyme did not work well, please inform someone.  While it might be attributable to a mistake on your part, it is also possible the enzyme is not working well, and it is a courtesy to inform co-workers of this possibility.  

 General Lab Cleanliness 

--          Following an experiment nothing should be left lying around the lab. Dirty glassware etc. should be rinsed and placed in the appropriate bin.  Gel boxes and other material that we wash ourselves should be thoroughly rinsed and placed on the sink to dry.  Once dry these items should be returned to the shelf and not left on the sink. 

--          Balances tend to become dirty easily; these should be checked and cleaned following each use. 

--          Periodically take the time to, e.g. clean the sinks, wipe out the incubators, remove extraneous glassware etc. 

Lab Tools 

You will need to make a few simple tools to be able to manipulate cells and embryos. Make up most of the tools in the first lab and then keep them for use throughout the semester. All of the tools can be sterilized before use with 70% ethanol.  

             Microknives –    One can use fine-tipped scalpels (expensive) or you can make your own using chips from double edged razor blades mounted on the ends of wooden dowels. Protective eye gear and leather work gloves must be worn when breaking the  razor blades.  

Break a razor blade into a number of small pieces by cutting it with heavy-duty scissors, aviation tin snips. Make a small slit in the end of a wooden swab and place the microknife in the slit. You can seal it in place with clear finger nail polish or with hot paraffin.  

            Microneedles - I normally make microneedles by cutting the blunt end off of # 00 or # 000 insect pins then sticking them in the end of a pasture pipette. Then seal in place by heating the end of the pipette/needle and touching it to a pellet of paraffin to hold  it in place. Finger nail polish can also be used to hold the needle in place.  

            Hairloops -        Make a loop of hair, yours or others, and place it in the end of a pasture pipette. Seal it in place with hot paraffin or nail polish. You should make several different size loops and if possible with different thickness hairs. These are used for manipulating eggs or embryo structures.  

            Eyelash Probes Pop out a couple of your eyelashes and insert them in the end of a pasture pipette. Seal in place with paraffin or nail polish. These can be used to move         embryos or structures in embryos. Because they taper to a fine tip, they are also  used as surgical tools to separate cells in early embryos.