Fortunately, our students and faculty are unequalled too! The Organizers and the Students are both mindful of this tremendous support and extremely grateful for it.

The course is unusual in many ways and I have written this guide so that everyone knows what is expected.

To create an environment in which students with interesting biological problems are brought together with advanced instrumentation in order to increase their understanding of the 3D Microscopy of Living Cells and to develop and disseminate solutions to the problems we find.

From the point of view of the manufacturer's representatives, this course is likely to involve more hands-on experimental effort than they have ever experienced in a one-week period. It may be exhausting, but it is also a valuable chance to learn first-hand about the problems faced daily by those who really push their instruments to the limit.

The following procedures have been developed over the years to make the Course a success, for everyone.

There is a Pre-course on the afternoon of Saturday, June 16. It runs from Noon to 5:00 PM and discusses and demonstrates basic optics. Manufacturers are not really involved in this.

Then, in spite of what you might have heard, the Live-cell Course proper actually starts with Registration, the Welcome Reception and Introductory lecture on the evening of Saturday, June 16. It is vitally important that all attend these events. We need you to introduce yourself to other members of your group and choose your group's SkitMeister.

The Course itself is presented in three parts: 2D (the first 5 days), 3D (the next 5 days) and Summary (the last day and a half). The first two sections are separated by the Manufacturer's Reception and the last two, (almost) divided by the Beach Party. After a one day break, the Live-Cell Course is followed by a 3-day workshop on 3D Image Processing (3D IP).

Each day starts at 8:30 with Announcements of problems and changes etc. These may involve you so don't be late! The morning is usually three "lectures," with a Coffee Break after the first one. We then all eat Lunch together. Except for the first two days, most afternoons are labs with a break for "tea," but during the 3D LAB we have only 2 morning lectures followed by Show and Tell where we share our results from the previous labs. Early in the course some evenings are used for lectures. Later during the 3D LAB they are used for cell prep and one lab session.

In general, Reps are expected to make sure that their instruments are attended during all scheduled labs and Personal Project periods (unless their instrument is not signed up).

On the other hand, outside of these times they are welcome to show their equipment to anyone else. Students not involved in Cell Prep may find time for a "personal introduction" during part of the hour before the scheduled afternoon 3D labs.

2D EQUIPMENT

Some 2D equipment (Microinjection) will be set up to coincide with the evening introductory lecture on this topic on Day 1 (June 17). Likewise, the laser-tweezers system will be functioning by the morning of June 18. Companies bring equipment for the time-lapse demos and should have it installed by the morning of June 18.

Those providing equipment for use in the 2D labs, on Days 3 and 4, will be warmly welcomed as soon as they arrive!!! It would be great to have some extra time to set up some long-term timelapse sequences.

The afternoon 2D labs (TIRF, Timelapse, µinjection, Laser Tweezers, Ion Imaging) are 2x2 hours and are held on Days 3 and 4 (June 19 - 20).

Those manufacturers planning to leave after the 2D course are encouraged to participate in the Manufacturer's Reception that is held the evening of Day 5 (Thursday, June 21).

3D EQUIPMENT (WF/decon, confocal, multiphoton)

3D equipment should be set up and ready to go by the afternoon of the fifth day of the course (Thursday, June 21 ). It should remain functional until at least 5:00 PM on the tenth day of the Course (Tuesday, June 26). However, there are usually many motivated students looking for 3D microscopes until late on the next day, and you are encouraged to keep equipment available for as long as is possible.

The Manufacturer's Reception is held on the evening of Day 5. Each manufacturer is given 5 minutes to make a very brief introduction to the MOST UNUSUAL new aspects of their instruments. The idea is to hit the high points for the student. The order of presentations is determined by lot.

After Announcements each morning, Reps are invited to attend both the lectures and food breaks. Food-breaks are great chances to meet and talk to reps. These contacts may be important when students come to make their choices of instrumentation to carry out your Personal Project in the evenings (more below).

The food is pretty good, too! See Manufacturer's Reception photos here.

For Reps only, there will be an introduction to the "Lab Projects of the Day" at 10:00 am (after coffee) on Days 6-11 (June 22-27). This is to help manufacturers be ready for the student projects when they arrive.

During 3D-LAB, the students have an hour after lunch to "look after their cells." At 2:00 PM, 3D-LAB starts. All students are part of a group of 3 or 4 who, we hope, share similar interests. Each student group has a specific Faculty or Super-volunteer Advisor. Before coming to the Course, they have all worked together to develop one or two Personal Projects that they hope to carry out during the course. So that Reps know what to expect, we are asking students to bring the protocols of the personal projects with them when they come.

There are 14 sessions of 3D-LAB (10 afternoon sessions and 4 evening sessions). To insure that all students get a chance on every instrument, each group will be assigned to a different 3D set-up for the first 10 sessions of 3D-LAB.

For 6 of these sessions (the afternoon sessions on the first 3 days: Friday-Sunday) , groups will work on projects assigned by the course. These projects have been designed to both work on the equipment available and to introduce students to important topics. The protocols for these assigned experiments will be available from the Protocols WWW page well before the course commences.

These are the protocols that will be discussed with the Reps at the 10:00 AM (after morning coffee) sessions, but one should also be able to access them from the WWW before the Course starts. Where it makes sense to do so (and where the change is compatible with the instrumentation available), Advisors may permit students in their group to make small modifications to these protocols (different cell type, different dye, etc).

Any changes need to be approved by the Advisor. However, the Rep is responsible for the safety of his/her equipment and is the final judge of what it can be used for. Towards the end of the 3D LAB, it is probable that more requests for changes will be approved.

During a 2-hour 3D LAB period, the Rep will have to balance how much priority is given to explaining the features of his/her instrument and how much is spent trying to get the experiment to work. As these experiments involve living cells, I suggest that Reps initially emphasize the cells. There will doubtless be breaks in the action when a cell dies or a preparation needs to be replaced. These breaks are ideal times for Reps to try to explain the intricacies to any students not totally involved in solving the problem.

As this is a "learn-by-doing" course, we encourage the Reps to guide a student operator rather than running the instrument themselves. Both you and they will learn more (and we encourage students to get into the driver's seat and "ask for the keys").

A word to the wise. Live cells do not like being looked at. All instruments should be aligned as perfectly as possible. On a confocal, you should use a larger than normal pinhole, and the "right" zoom setting (not too high). All microscopes should have a functional cell chamber (with culture dishes if required) and water lens and you should have read, and understood, the tutorial on Spherical Aberration (a big topic this year!!!) posted on the WWW site, so that you can properly adjust the correction collar on the water lens that you should have brought!!!

The best results are likely to come from those instruments that collect useful images while subjecting the cells to the least possible laser (or other) light load (ND 2 or 3 filters on the laser are normal). Reps should practice this ahead of time. The Organizers have a laser power meter, if you wish to check power levels at the specimen. It should be in the low microwatt range.

To press this point home, the first assigned lab session will involve getting the best possible image, AFTER 10 scans, of a very bleachable specimen. Reps can try this ahead of time.

Reps who can deconvolve or apply a Gaussian filter to Nyquist-sampled data, will be able to remove a lot of Poisson noise, although, they will also have to increase the display contrast to see much (see ch 19).

It is inevitable that, on a given day, some 3D LAB projects will be more successful than others. So that all may taste success (and learn from defeat!!), on each day, the period just before lunch is reserved for all to see what was produced the previous day. Data files (usually in the form of movies) are transferred on CDs or DVDs disks to computers in the lecture room and students say a few words about what was done to produce the results projected onto the screen. Technical details of the experiment should be filled in on a table on the blackboard. Please don't bring files so huge they take an hour to read!

This is NOT a competition. It is an attempt to ensure that we learn as much as we can from what goes right.

The accepted response to the problems that inevitably occur is to try to help solve the problem. Far from pointing fingers, I have often been heartened by the generous assistance provided by alleged commercial competitors.

Students should not ask the Reps to comment on equipment that they do not sell. They can make "critical" comments about their own equipment but not that of others!

Encourage the reps to attend all SHOW-AND-TELL sessions. They will not only be able to ensure that results obtained on their instrument are displayed to their best advantage, it is also very likely that they may learn from the discussion and from the results produced on other instruments.

After a break for dinner, students return for their evening sessions. The period from 7:00 to 9:30 is divided into a half hour prep sesson and a 2-hour lab. Groups go to specific instruments that have either been assigned (first 3 nights of 3D section) or which they have signed up for, starting the previous Noon (noon on the previous day). The "Sign-up Sheet" will be in the Lunch Room (usually on the blackboard).

To prevent the "Trampling of the Slow," there are some limitations on the instruments that a group can initially sign up for. In particular, groups should not initially sign up for an instrument that they have already used.

The reason for the "day-early" sign-up is to give as much warning as possible to the manufacturers and to allow you the time to plate the cells you need onto a substrate compatible with the appropriate cell chamber.

It also leaves enough time for a second, OPEN SIGNUP period between NOON and 5:00 PM of the day of the time slot. During OPEN SIGNUP, any group, or part of a group, may sign up for any instrument not already reserved. Therefore, by 5:00 PM the Reps will know if and when they will have to attend their system that evening.

We usually organize the 28 - 32 students into 8 groups and have 10-11 3D microscopy workstations operational. To resolve this mismatch, we may organize more groups (maybe 10?). Alternatively, we may allow groups to divide to utilize unused systems.

There is still a problem: the 3D workstations are only available for 5 days. We assign groups to specific instruments for the first 10 sessions of 3D LAB. Although 10 is not 11, some instruments are quite similar. We hope that on the remaining periods of 3D LAB, the "can't use scopes you have used before" Rule will insure that everyone has a chance to see everything.

There is no evening lab on the night of the Beach Party and on the last four sessions of 3D Lab, there is a lot of horse-trading between groups about who gets what, when. Although not ideal, in general, I feel that this system is the one most likely to lead to the "most good for the most people."

Weather permitting, it is held on the evening of the ninth day (Monday, June 25). This is a turning point of the Course. It is the time when students will come to realize that they may not win a Nobel Prize with the results they can get this week! The Party is a punctuation mark that is placed here to prevent ever-more-frantic efforts to "save the day" (and missing lectures!) from destroying any chance of "learning anything" during the remainder of the Course.

The Beach Party should help you start thinking about what you CAN accomplish in the remaining time: One more full day of 3D LAB, then a day to "Massage the Data" before the Student Presentations on Day 12.

To lure everyone from the bench to the lunch, the Course spends a small fortune on a Feast of Fresh Seafood (Salmon, prawns, sacks of oysters (literally: fresh and baked), barrels of clams, corn and salads and pies and fruit.) as well as chicken, ham and veggies for those who prefer them. The Party provides this feast in a matchless setting: a lonely beach from which one can see both downtown Vancouver and the apparently deserted islands of Georgia Strait. The sunsets are often memorable.

We also award a number of Golden Salmon Prizes to the winners of contests, designed to take one's mind off things micro:

Best Skit Contest:

Students' impression of arrogant/disorganized faculty. (Each groups will have chosen a SkitMeister as part of the Welcome Reception on Day 0). We have yet to have a manufacturer win but they are invited to try.

Faculty Slide Contest:

Groups will sketch possibly ambiguous diagrams on posters. Faculty must utilize these as part of a 2 minute lecture on topics microscopical. Contest judged by Reps on site.

Beachwood Confocal Contest

Groups fabricate, and describe the operation of, fantastical microscopes made out of things found on the beach. Contest judged by Reps on site.

Crazy Advertisement Contest

FINALLY! THE REPS GET A CHANCE TO MAKE UNRESTRAINED PITCH!!!

Reps are allowed 3 minutes to hawk, with unparalleled pushiness, an imaginary product, (very) distantly related to the 3D microscopy of living cells, while looking out through the hole in a large, cardboard "TV set." (past examples include: CD of Lullaby's for Cells?, Make-up for the fashionable microscopist? Breath deflector so that bad breath doesn't disturb cells? Cell Sunscreen? You get the idea.) Points given for applause, imagination and enthusiasm!

On the last morning of the Course, we take a moment to go over what we have accomplished. We do this by giving each group 15 minutes to describe the successes they have had with their Personal Projects.

The results are really remarkable, and a tribute to the efforts of all involved!!

Then, we have lunch and say goodbye!

There is then a rest day before starting the 3D Image Processing Workshop