*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
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 <[log in to unmask]> 3/19/2013 12:14 PM >>>
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
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
[log in to unmask]
privatewww.essex.ac.uk/~plaissue
On Tue, Mar 19, 2013 at 1:23 AM, Arvydas Matiukas <[log in to unmask]>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.: 315-464-7997
> fax: 315-464-8014
> email: [log in to unmask]
>
> >>> 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
> [log in to unmask]
> 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
> >
>
|
