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I am a PhD stuent from University of Dlehi, India. Currently I am
performing some experiments with mito tracker and ER tracker and was
studying mitochondrial movement towards ER. When i tried co localization i
was nt happy with my results. I am using 10 nm of mitotracker & er tracker.When
i looked into literature i got sm pictures like what i hv got,
but I found that they further digitally magnified the images several fold.
I tried with the photoshop & imagej to get the best magnificatn. But still
nt much convincd. Any suggestn please.
Regards
Chaitali Banerjee
PhD student
Department of Zoology
University of Delhi
India


On Mon, Feb 24, 2014 at 8:45 PM, Glen MacDonald <[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
> *****
>
> Given the gmail accounts and lack of affiliation, are you and Chaitali
> working together?  Are you looking for signal or resolution?  Do you have a
> suitable camera?  An overly corrected objective, low NA or phase objectives
> will all reduce signal.  Mitotrackers in zebrafish can be detected in
> widefield or confocal with a 40X/.75 (or 20X + 2X zoom) air lens, and
> easily recorded with NA 1.0-1.2.  I hope you are not trying to look through
> a plastic culture dish.  Do you have any experience with live imaging,
> microscopy or dye loading? Have you looked in the literature?   There are
> many papers using Mitotrackers, and in zebrafish.
>
> Providing a description of your instrumentation, experimental intent and
> sample preparations when  you ask for assistance will avoid a game of "20
> Questions".
>
> Mitotrackers require testing for concentration, incubation times,
> temperature, etc. before embarking on any experiment but the literature
> will provide starting points.  Overloading can reduce intensity (by
> disrupting mitochondrial function), and is a common beginner's mistake.
>  try 25 nM to 100 nM, 20-30 min. or shorter.  MT Red is brighter, use no
> more than 25 nM, Deep Red behaves like MT Green, bt possibly a bit voltage
> sensitive.
>
> Digital zoom will not improve signal, it sometimes helps with visibility
> on the screen.
>
> Glen MacDonald
>         Core for Communication Research
> Virginia Merrill Bloedel Hearing Research Center
>         Cellular Morphology Core
> Center on Human Development and Disability
> Box 357923
> University of Washington
> Seattle, WA 98195-7923  USA
> (206) 616-4156
> [log in to unmask]
>
>
>
>
>
>
>
> On Feb 24, 2014, at 3:06 AM, Amandeep Walia <[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
> > *****
> >
> > Dear even i am trying to but getting nothing
> >
> >
> >
> > On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee <
> > [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
> >> *****
> >>
> >> I am also trying to localize mitochondria in fish macrophages using
> >> mitotracker. What I am getting is greenish tinge of mitotracker at x 100
> >> objective. Can u please suggest me if there is some software for digital
> >> zooming
> >> Regards
> >> Chaitali
> >>
> >>
> >> On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[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
> >>> *****
> >>>
> >>> Could you please tell what exactly you are going to image and what is
> the
> >>> label?
> >>> Are you going to perform some quantitative imgaing or just make sure
> that
> >>> your
> >>> sample really contains mitochondria? What is the cell type?
> >>>
> >>> Mitchondria are well visible for 1.2 NA objective (TMRE staining,
> >>> lymphocytes) and
> >>> qualitatively visible while using NA 1 objective if cells are spreaded
> >>> (MitoTracker
> >>> Green, Vero cells)
> >>>
> >>> Best
> >>> Sergey Tauger
> >>> PhD student
> >>> Cell motility lab.
> >>> Dept. of Biology
> >>> Moscow State University
> >>>
> >>
>