Re: Advice for confocal training

[phil laissue-2]

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Hi Arvydas,

just wanted to add to this old thread that I too have a 'fast-track' route
which takes about three hours, and for some users, that's perfectly okay, I
agree. As for stage crashing: It's lens crashing, really, but the Nikon
objectives are so chunky and the z-motor so powerful, they bent the stage
bracket out of shape. Happened with those LabTek chamber slides, trying to
image the ones on the far end.
Re training: Eleanor Kable presented this nifty approach at FOM:
http://www.ammrf.org.au/myscope/
So one thing you can do is ask users to peruse it, take the test, then
bring the certificate to the practical lesson. They apparently really
appreciate being able to go through it at their own pace, and like to see
if they understood correctly by answering questions. I'll certainly have a
go at it with my users.

Kind regards

Philippe


_____________________________________
Philippe Laissue, PhD, Bioimaging Manager
School of Biological Sciences, Room 4.17
University of Essex, Colchester CO4 3SQ, UK
(0044) 01206 872246 / (0044) 07842 676 456
[hidden email]
privatewww.essex.ac.uk/~plaissue


On Tue, Mar 19, 2013 at 8:56 PM, Arvydas Matiukas <[hidden email]>wrote:

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>
> Hi Philippe,
>
> thanks for the details how you train your users. My experience
> is very similar to yours  that a new user requires about 10hrs of
> training/work with confocal before he/she can start productively
> work on their imaging project independently. On the other
> hand about 3hrs of  practical training (i.e showing which buttons
> to push and which icons to click) is fairly enough to prevent
> users from breaking the equipment. Most of my users assume
> that this is enough of me wasting their time and typically
> prefer as a first choice to ask their previously trained coworkers which
> buttons
> to push. I kind of understand their quick pace which (as you wrote) often
> is imposed
> by a lab leader requesting quick results. Moreover, usually sample
> preparation
> takes much longertime than imaging and still does not guarantee good
> picture.
>
>
> Unfortunately, until now our campus did not have theoretical
> course on bioimaging/microscopy but I am working hard to change
> this. It that respect your experience is very useful.
>
> Finally, I wonder how it is possible to crash a stage (never saw or heard
> anything of such kind). Our inverted Axiovert 200M has a button to set the
> top
> limit of objective advancement which protects from crashing lens. However,
> the caveat is if somebody pushes that button at wrong z-position (while
> objective
> is outside working distance) then it is not possible to focus without
> restarting
> the microscope power.
>
> Best,
> Arvydas
>
>
> >>> phil laissue <[hidden email]> 3/19/2013 12:14 PM >>>
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> To join, leave or search the confocal microscopy listserv, go to:
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>
> Hi Arvydas,
>
> thanks for the feedback. I run four lectures (basics, dynamic range,
> colour, 3D & time-lapse) of 1.5 to 2 hours each (depending on questions) at
> the start of each term. I also have a three-day course each year with four
> 1.5h lectures followed by practicals. The third day is used entirely for
> samples the users have prepared. A strong point of the lectures is the
> practical aspect. I talk about the focal planes, lazy oil, and teach them a
> standard procedure to find their cells. This sounds very pedestrian, but
> I've seen often how PhDs spend half their time at the 'scope looking for
> their cells. We've also had two crashed stages (but luckily no objectives
> were hurt) in the first half year of having the new instruments, and the
> new way avoids that. It would be good to have parking sensors on stages. I
> still have to figure out if it can be done software-wise with NIS-Elements.
> I fully agree that many users could possibly care less, but not by a very
> large margin. I run the courses with interested students in mind, but also
> have the lectures on a training website, so they can be downloaded and
> studied in advance of a practical session. Printouts and crib sheets
> complement that. What does help with motivation is to make students
> understand that bioimaging is a hot area and increasingly desired on CVs.
> And so I end up with about one third interested, good, careful users, one
> third routine (but responsible) users, and one third of intermittent (and
> rather bored) users.
> David Piston, and John, are of course right. Rigorously speaking, you can
> probably only truly understand (and appreciate) a system if you've built
> one yourself. But multi-user facilities with all members having such
> insight and skills must be extremely rare. For many, it's just an
> instrument they want to use. So to some degree, I have the lectures for
> myself. Just showing the users which button to push is not fun. However, in
> defense of the students, it seems that the pressure and lack of
> understanding for the complexity of the matter (or the time it requires to
> do it properly) comes in some cases from the group leader. So another point
> I put forward is that it only makes sense to embark on a bioimaging project
> if they're willing, ready and able to put in the time; sample optimisation,
> proper image acquisition, and image analysis being aspects that can swallow
> a lot of time. But there's no 'quick & dirty' way to do it really, unless
> all they want is a pretty picture for a publication. So the minimum time
> charged for confocal training is 10h. Alison North's article (Seeing is
> believing? A beginners' guide to practical pitfalls in image acquisition)
> really helps to drive that point home as well. And when a user gets the
> imaging bug, all that effort seems worth while...
>
> Best regards
>
> Philippe
>
>
> _____________________________________
> Philippe Laissue, PhD, Bioimaging Manager
> School of Biological Sciences, Room 4.17
> University of Essex, Colchester CO4 3SQ, UK
> (0044) 01206 872246 / (0044) 07842 676 456
> [hidden email]
> privatewww.essex.ac.uk/~plaissue
>
>
> On Tue, Mar 19, 2013 at 1:23 AM, Arvydas Matiukas <[hidden email]
> >wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hi Philippe,
> >
> > I am impressed by the content and scope of your general lectures.
> > I wonder how many hours they take. I wish I  could implement
> > something similar, however, I am 99% positive that biomedical grad
> > students that are typical users will get bored to death. On the
> > other side, without a  proper prerequisite of math/physics/optics
> > users woudn't be able to in depth grasp the basic microscopy
> > elements.
> >
> > Regarding Jon's question I would like to suggest to use Zeiss or
> > Leica's confocal training presentations (just google them).
> > They contain a well balanced mixture of theory/practise/applications, and
> > take about 3-4  hrs to cover.
> > Regarding training cost, basic trainingin my Core  is below $100 ( the
> > policy is
> > to atract a new customer without scaring him/her with high price).
> >
> > Best regards,
> > Arvydas
> > ---------------------------------
> >
> >
> >
> > Arvydas Matiukas, Ph.D.
> > Director of Confocal&Two-Photon Imaging Core Facility
> > Department of Pharmacology
> > SUNY Upstate Medical University
> > 766 Irving Ave., WH 3159
> > Syracuse, NY 13210
> > tel.: [phone]
> > fax: [phone]
> > email: [hidden email]
> >
> > >>> phil laissue  03/18/13 8:40 PM >>>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hi there,
> >
> > it really depends on the targeted users - needs differ much depending on
> > the place/lab. I tailor practical sessions usually to individual needs,
> but
> > have general lectures at the start. Basic lectures include: when to use a
> > confocal, dynamic range, histograms, saturation, pixels, LUTs + real
> > colour, basic filter settings, bleed-through; z-stack, zoom, Nyquist,
> > step/pinhole/image sizes, scan speed, laser power vs detector gain, SNR,
> > averaging/summing, considerations for experimental setup and live cell
> > imaging, sample quality control; PSF deconvolution, visualisation
> > (tiling/projections, scalebars, contrast) and preparing figures/movies
> for
> > presentation/publication. Basics in practicals include finding your cells
> > without crashing the lens (for automated XY stages), adjusting channels,
> > bleaching, taking a z-stack. We don't have too many users wanting
> advanced
> > techniques, so I keep information on TIRF/spectral/resonant/FRAP et
> > al./advanced image analysis options to a 'this is what it can do' level,
> > and get into details if they want to/need to use it.
> > For basic use, I hear that the LSM 700 does the job, but myself would
> > consider a Thorlab kit (although that again depends on the user basis) or
> > second-hand model (510, SP2, Radiance in good shape). I'm happy with our
> A1
> > workhorse.
> > Samples depend on the users in question, but transiently transfected
> cells
> > with varying intensities, strongly autofluorescent samples, diatoms,
> beads,
> > bulky whole mounts and poor slides (e.g. with air bubbles or poor signal)
> > all come in handy to make certain points...
> > I'm sure I forgot a few things, but hope this helps for starters.
> >
> > Kind regards
> >
> > Philippe
> >
> > _____________________________________
> > Philippe Laissue, PhD, Bioimaging Manager
> > School of Biological Sciences, Room 4.17
> > University of Essex, Colchester CO4 3SQ, UK
> > (0044) 01206 872246 / (0044) 07842 676 456
> > [hidden email]
> > privatewww.essex.ac.uk/~plaissue
> >
> >
> > On Mon, Mar 18, 2013 at 9:56 PM, jmkrupp jmkrupp  wrote:
> >
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > *****
> > >
> > > Greetings
> > >
> > > Just a quick note to ask if anyone would like to give me some advice
> > about
> > > getting started with a confocal training program.
> > >
> > > We do EM and LM already, thinking of adding LSCM. Any advice about must
> > > includes,  good practice specimens , techniques etc?
> > >
> > > How about insights into instrurments, how basic can we go?, what cost
> > range
> > > should we be thinking.
> > >
> > > Thanks
> > >
> > > Jon
> > >
> >
>