Hi Valeria
I have never imaged mouse
embryos but only mouse skin and zebrafish and C.e. embryos. However it seems to
me that if you want to follow cells in a mouse embryo, a 2-photon system would
be best as you can image rather deep. We find that to follow labelled cells
among non labelled cells and know what is going on, it is crucial to acquire an
excellent DIC image. I would recommend oblique illumination (Dodt contrast) which
is available with several commercial 2-photon systems. I would also say that if
you have no need to image intracellular events, an excellent 20x objective will
work just fine for your purpose while giving you the field of view and the NA
needed.
Med vänlig hälsning / Best regards
Sylvie
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Sylvie Le Guyader
Live Cell Imaging Unit
Dept of Biosciences and Nutrition
Karolinska Institutet
office: +46 (0)8 608 9240
mobile: +46 (0) 73 733 5008
From:
Sent: 21 June 2010 17:19
To:
[hidden email]
Subject: Embryo time lapse
Dear all,
I am running a facility with many different users and I am trying to pull
together their needs on imaging in order to improve the equipments.
Right now the biggest demand is about time lapse imaging (long term time lapse
(24-48hours with a 15-30min step)) specifically on mouse embryo (E7-E8) with
fluorescence (GFP,Venus,dsRed).
So we need very high sensitivity system, weak photo damage, both high
magnification (to follow single cells) and possibility to follow movement in the
entire embryo.
Too many tasks for a single system?
Which do you think would be the best system to achieve these goals?
a confocal? a multiphoton? the new confocal-stereomicroscope from Nikon AZ-C1?
a regular inverted widefield with a good camera?
Which system do you use for these
time lapse?
Thanks for all your always helpful
input
Valeria
-- Valeria Berno,PhDMicroscopy Facility ManagerMRC Centre for Regenerative MedicineInstitute for Stem Cell Research