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March 2013

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Confocal Microscopy List <[log in to unmask]>
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phil laissue <[log in to unmask]>
Date:
Tue, 19 Mar 2013 00:39:32 +0000
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*****
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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 <http://privatewww.essex.ac.uk/%7Eplaissue>


On Mon, Mar 18, 2013 at 9:56 PM, jmkrupp jmkrupp <[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
> *****
>
> 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
>

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