Developing a course on microscope use

[Rorschach]

Hi people,

I've started to develop an idea for a comprehensive course on microscopy, how to use them properly, develop a practical understanding of what can be achieved using different microscopes and contrast techniques and how. Also a segment on how to prepare samples and specimens. Importantly, also a segment on proper maintenance and cleaning of scopes. Basics of microphotography would form one segment.

At this stage, when I need to market the idea to a couple potential funding agencies, it would be extremely valuable to be able to show a series of images of the same specimen shown using different contrast techniques: brightfield, darkfield, phase contrast, DIC (if possible, fluorescence too). Ideally the specimen would be in the exact same orientation with no change other than the contrast technique being used. Can you point out a net resource where that could be found?

The course would take place in my own university in Finland and the target audience would be PhD students from several different departments and faculties.

Electron microscopy and confocal microscopy etc. are ruled out as being too challenging, plus there are no hardware for those. So the covered types/techniques would be 1) stereo microscopy (BF, DF, polarized and Rottermann contrast and, obviously, reflected light) and 2) diascopic microscopy (BF, DF, phase, DIC and fluorescence) for both transmitted and reflected. I'll describe the existing and planned equipment for these later today in another message.

If successful in getting funding, I may be looking for a couple enthusiastic teachers who know these topics Travel and accommodation costs would be covered as well as a reasonable commission. Duration would be 5-7 days and venue is in the middle of one of the nicest national parks in Finland!

Anyone interested?

Best regards,
Riku

[TonyT]

some
HERE

[Rorschach]

some
HERE

Thanks Tony! Those are exactly what I had in mind. Won't hurt to find more, though.

I would need permission to use these of course (naturally with the owner/photographer mentioned in each shot), even at the funding proposal stage. Proposals usually aren't publicly available but I am pretty adamant about copyright and usage rights issues, even for educational purposes. Plus the photos would be also needed later for an ad about the course and that would obviously be online in several places. Provided that I am able to pull this off!

[TonyT]

Your welcome to use them, use ©AW Thomas

May be a good idea to rotate the full-sized image (if needed) to correspond to the leg images

[Rorschach]

So the equipment that will be available for the course consists of the following:

1 Stereos
1a) A Zeiss StereoDiscovery.V20 with Plan Apo optics, including an objective revolver with Plan Apo S 0.63x and Plan Apo S 1.5x, motorized focus & zoom, Z-stacking using Zen Blue, LED cold light (BD, DF, polarized transmitted & slit, ring reflected), a Zeiss 4K camera connected to a 4K Sony projector. Plans in motion for getting a Zeiss equivalent of a Rottermann Contrast base for it.
1b) A Leica 205C manual stereo with an objective revolver (Plan Apo 1x & Plan Apo 2x corr.), AX carrier (can't be used with the revolver though), a Leica BF, DF & Rotterman Contrast base, Photonic led spots for reflected. It has a full HD Leica camera at present but we are planning to upgrade that. Will also be connected to the 4K projector.
1c) Altogether ten Zeiss Stemi 2000 and 2000c, all with the new Stand K Edu (6 units) or the Stand K Lab (4 units). These have reflected LED lights and BF & DF transmitted. One Stand K Lab has polarization equipment.

2 Diascopic scopes
2a) A Reichert Polyvar bio is being purchased this year. It will have BF, DF, DIC and polarized for transmitted. Maybe epifluorescence too, let's see. A LED conversion and a modern (maybe Zeiss) microscope camera also coming (connected to the 4K projector).
2b) Our Olympus IMT-2 will be upgraded to DIC and polarized, it already has BF, Fluorescence and phase. It will also receive a LED conversion (not the fluorescence system, though). Also a modern microscope camera for it, maybe a Zeiss MrC.
2c) An Orthoplan equipped for reflected (BF, DF, DIC, simple polarization) and a LED conversion. A Zeiss camera will also be installed.
2d) a Zeiss Axio AxioScope.A1 equipped for BF & phase, with a camera
2e) An assortment of smaller diascopic scopes for student use, including two relatively new Zeiss scopes but also older Leitz, maybe even a couple Wild M20. Most of these have BF, DF and phase. Total number will be 5-7.

We also have a Leica microtome that will be set up for use. The plan is to do the three mentioned LED conversions using Nanodyne kits, unless a better alternative comes up. Any experience on them or hints otherwise?

Of course, the venue being a research station, we have a huge pile of different sampling gear at our disposal plus various traps, boats and field gear.

[Phill Brown]

Inverted scope and sample cleaning and preparation?

[Rorschach]

Inverted scope and sample cleaning and preparation?

Actually, the Olympus IMT-2 is an inverted scope. But sample cleaning and preparation is an important topic to cover.

Or did you specifically mean sample preparation for use on an inverted?

[Phill Brown]

Inverted scope and sample cleaning and preparation?

Actually, the Olympus IMT-2 is an inverted scope. But sample cleaning and preparation is an important topic to cover.

Or did you specifically mean sample preparation for use on an inverted?

I would be specific for sample prep for inverted.

[Rorschach]

Inverted scope and sample cleaning and preparation?

Actually, the Olympus IMT-2 is an inverted scope. But sample cleaning and preparation is an important topic to cover.

Or did you specifically mean sample preparation for use on an inverted?

I would be specific for sample prep for inverted.

Yes, I agree. It's rather different from other sample prep.

[apochronaut]

This is always a good idea. To what magnification do the stereos take you?

[Rorschach]

This is always a good idea. To what magnification do the stereos take you?

The StereoDiscovery.V20 goes to 225x but we also have the PlanApo S 3.5x mono attachment for it, which takes mag to 525x using the 10x/23 eyepieces. Probably quite a bit of empty mag when you go that high. On the revolver, there's also an axial position for both objectives so with the motorized focus & Zen Blue z-stacking software, this scope is absolutely great for microphotography.

The Leica M205C goes to 320x as we have also a PlanApo 2x on the revolver. That PlanApo 2x is the rare special 'corr'' version with a dial to take into account the depth of water or ethanol (which have nearly identical refractive indexes that differ a lot from that of air) over the specimen, bringing an improvement in resolution in these situations: https://www.directindustry.com/prod/lei ... 10450.html The M205C employs Leica's Fusion Optics concept which brings a slight improvement in perceived depth of focus without losing resolution: https://www.leica-microsystems.com/scie ... al-images/
So, on the big Leica we have top notch optical quality like the big Zeiss but additional goodies converging: the fusion optics, the Plan Apo 2x corr. and Rottermann contrast...it's a killer scope for benthic macroinvertebrate id work or, well, anything in liquid.

The Stemi 2000/c go to 50x. Their recently bought new stands Stand K Edu and Stand K Lab make a big difference in terms of visual clarity and contrast, also usability. The spot, DF and BF lights are super good. In every respect, a huge difference with the original stands they had. The only down side is that they lack fine focus.

Edit: corrected a typo. the M205C mag goes to 320x

[apochronaut]

I have struggled over the years to bridge that gap between a high quality stereo and the bottom end of a diascopic microscope. Recently, I ponied up to bridge that gap up to 135X. Looks like you have that covered well.
This new generation of planapo stereos are quite something.

[wabutter]

You may want to look into the Microscopy courses offered at Woodshole, Mass and Cold Springs Harbor, in New York. These are high level microscopy courses with extensive support from the big four manufacturers including manpower to support workshops and instrumentation. Of course, this give the companies exposure to highly interested individuals. Courses, if I remember correctly ran for 7-10 days and people paid tuition to attended as well as their own expenses. I don't know if anything like this is available in Europe, but it would certainly give a bench mark to work from.

[SuiGenerisBrewing]

You may want to look into the Microscopy courses offered at Woodshole, Mass and Cold Springs Harbor, in New York. These are high level microscopy courses with extensive support from the big four manufacturers including manpower to support workshops and instrumentation. Of course, this give the companies exposure to highly interested individuals. Courses, if I remember correctly ran for 7-10 days and people paid tuition to attended as well as their own expenses. I don't know if anything like this is available in Europe, but it would certainly give a bench mark to work from.

There are a number of great microscopy courses offered in Europe, although most of the ones I know of target confocal and super-resolution approaches - that's not to say that there aren't other courses; those are just the areas I work in. The Royal Microscopical Society offers several good courses - UK based, of course.

For the OP - I've run a number of microscopy courses covering a range of methods. I teach basic staining, brightfield (+ phase contrast and darkfield), and fluorescence to undergrad students as part of a microbiology lab course I run, I run a small microscopy core facility where I frequently teach a range of widefield fluorescent approaches (basic labeling through to FRET + single molecule approaches), plus I've taught advanced fluorescence and super-resolution approaches in national (Canadian) courses. A few unsolicited pieces of advice, based on that experience:

• You need to build your course around your target audience. A course effective for getting new users up-and-running with basic approaches needs to be very different from a course teaching experienced microscopists new/advanced approaches.
• Consider limiting the fields that you draw students from. It is quite hard to teach microscopy to a group with people interested in a broad array of use-cases. I once taught a course to a group consisting of individuals interested in everything from biomedical imaging, to minerology, to metallurgical microscopy...it did not got well.
• Even with advanced users, don't assume they know basic theory, light paths, sample prep, etc. It is amazing how many people with years of experience in a technique only know that technique by rote learning, and are unable to explain the fine details of the method or how to troubleshoot when it goes wrong. With advanced users, we always do a 30-minute crash-course on microscope basics. For new trainees, that same material gets taught over several modules.
• Many users (new and experienced) often fail to make the link between how a sample needs to be prepared is dependent on the microscope and imaging modality you want to image that sample with. You have a great set of instruments - I'd suggest linking each instrument with a sample preparation approach that gives a great sample for each scope. This allows you to build proper sample preparation into the use of the microscope, you can use the sample preparation time to also explain what makes the scope unique and why samples need to be prepared in a certain way for that scope, and then the trainees can see the samples they've prepared on the microscope itself.
• You can save a lot of time in the course by having short and simple on-line modules that the trainees complete in advance of the course. We use this approach to teach a lot of the theory (image formation, Nyquist sampling, light paths, how phase contrast/darkfield/etc works, Kohler alignment, etc) prior to our courses. This allows you to focus the in-person classroom time on sample preparation, microscope use, etc, rather than on theory and background.
• Include some time for teaching and performing image analysis. Again, this is a microscopy skill that even experienced users often lack, but which is critical for making the most of microscopes.
• While a PITA to prepare, having a printed manual/workbook for the course can be highly beneficial. For new users in particular, having a workbook with protocols, work sheets, and places to record notes/data, really helps the trainees work their way through the different modules in the course, and to centralize the information they are learning.
• Reach out to the microscopes manufacturers (especially anything you've purchased lately). They are often willing to send sales or technical support people (sometimes even with fancy microscopes) to teach in your course. This gets another expert in the room, can bring in new/exciting instruments for you (and the trainees) to play with, and they generally are free. They are especially willing to do this if the course is held in a city where they already have a resident sales/technical support person.

Bryan

[Rorschach]

You may want to look into the Microscopy courses offered at Woodshole, Mass and Cold Springs Harbor, in New York. These are high level microscopy courses with extensive support from the big four manufacturers including manpower to support workshops and instrumentation. Of course, this give the companies exposure to highly interested individuals. Courses, if I remember correctly ran for 7-10 days and people paid tuition to attended as well as their own expenses. I don't know if anything like this is available in Europe, but it would certainly give a bench mark to work from.

Thanks for the tip. Valuable info on the structure of the course and other things. They are very much life-science dominated, though. Our target people would be earth sciences: ecology, geography, maybe some from geology and water & environmental engineering.

[Rorschach]

You may want to look into the Microscopy courses offered at Woodshole, Mass and Cold Springs Harbor, in New York. These are high level microscopy courses with extensive support from the big four manufacturers including manpower to support workshops and instrumentation. Of course, this give the companies exposure to highly interested individuals. Courses, if I remember correctly ran for 7-10 days and people paid tuition to attended as well as their own expenses. I don't know if anything like this is available in Europe, but it would certainly give a bench mark to work from.

There are a number of great microscopy courses offered in Europe, although most of the ones I know of target confocal and super-resolution approaches - that's not to say that there aren't other courses; those are just the areas I work in. The Royal Microscopical Society offers several good courses - UK based, of course.

For the OP - I've run a number of microscopy courses covering a range of methods. I teach basic staining, brightfield (+ phase contrast and darkfield), and fluorescence to undergrad students as part of a microbiology lab course I run, I run a small microscopy core facility where I frequently teach a range of widefield fluorescent approaches (basic labeling through to FRET + single molecule approaches), plus I've taught advanced fluorescence and super-resolution approaches in national (Canadian) courses. A few unsolicited pieces of advice, based on that experience:

• You need to build your course around your target audience. A course effective for getting new users up-and-running with basic approaches needs to be very different from a course teaching experienced microscopists new/advanced approaches.
• Consider limiting the fields that you draw students from. It is quite hard to teach microscopy to a group with people interested in a broad array of use-cases. I once taught a course to a group consisting of individuals interested in everything from biomedical imaging, to minerology, to metallurgical microscopy...it did not got well.
• Even with advanced users, don't assume they know basic theory, light paths, sample prep, etc. It is amazing how many people with years of experience in a technique only know that technique by rote learning, and are unable to explain the fine details of the method or how to troubleshoot when it goes wrong. With advanced users, we always do a 30-minute crash-course on microscope basics. For new trainees, that same material gets taught over several modules.
• Many users (new and experienced) often fail to make the link between how a sample needs to be prepared is dependent on the microscope and imaging modality you want to image that sample with. You have a great set of instruments - I'd suggest linking each instrument with a sample preparation approach that gives a great sample for each scope. This allows you to build proper sample preparation into the use of the microscope, you can use the sample preparation time to also explain what makes the scope unique and why samples need to be prepared in a certain way for that scope, and then the trainees can see the samples they've prepared on the microscope itself.
• You can save a lot of time in the course by having short and simple on-line modules that the trainees complete in advance of the course. We use this approach to teach a lot of the theory (image formation, Nyquist sampling, light paths, how phase contrast/darkfield/etc works, Kohler alignment, etc) prior to our courses. This allows you to focus the in-person classroom time on sample preparation, microscope use, etc, rather than on theory and background.
• Include some time for teaching and performing image analysis. Again, this is a microscopy skill that even experienced users often lack, but which is critical for making the most of microscopes.
• While a PITA to prepare, having a printed manual/workbook for the course can be highly beneficial. For new users in particular, having a workbook with protocols, work sheets, and places to record notes/data, really helps the trainees work their way through the different modules in the course, and to centralize the information they are learning.
• Reach out to the microscopes manufacturers (especially anything you've purchased lately). They are often willing to send sales or technical support people (sometimes even with fancy microscopes) to teach in your course. This gets another expert in the room, can bring in new/exciting instruments for you (and the trainees) to play with, and they generally are free. They are especially willing to do this if the course is held in a city where they already have a resident sales/technical support person.

Bryan

Thanks Bryan. Many valuable points in there. Likely our target audience will mostly be not very advanced people in terms of their microscopy experience and skills. So that will have an influence on many things. Also, we are not going to target the biomedical or metallurgical segments at all, as those groups already have their own microscopy teaching in their departments and faculties, using very different instruments (modern cutting edge, including electron microscopy, confocal etc.). I like the idea of linking an instrument mostly to one sample preparation workflow and mainly one contrast method, especially as that means less changing of equipment back and forth on the scopes. On the Polyvar and Vanox for example it's not possible to do phase and DIC on the same scope unless you change lenses. So, we could have the Zeiss Axioscope for phase, Polyvar for DIC and darkfield and Vanox and/or Orthoplan for brightfield and simple polarization. Regarding fluorescence I am not sure yet. Depends on a few things whether it is possible to have it onboard or not.

Both of you bring up the manufacturers and that is certainly an interesting option. However, they might not be all that enthused about the fact that most of our diascopic equipment will be old legends of the golden era 30-40 years ago Stereos are current though, so maybe Leica or Zeiss could be enticed to send someone.

[wabutter]

All comment from SuiGenerisBrewomg are spot on. His last point about reaching out to the manufacturers is extremely valuable.

For courses that are more industrial and geological. McCrone Research In Chicago offers many course in quantitative microscopy especially using polarized light microscope. For many years alternating in offering the Intermicro meeting between the US and London.. However, I don't think it has been in London for a number of years.