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Sorry, it wasn't meant for all of you.
A

Andrea Manazza, MD PhD
Dept. of Biomedical sciences and human oncology
Molecular Oncology-CeRMS
Via Santena 7, 10126 Torino
Italia
tel: +39-11-6336859
fax: +39-11-6336887


>
>
> -------------------------- Messaggio originale ---------------------------
> Oggetto: Re: leading edge tracking
> Da:      "Andrea Manazza" <[log in to unmask]>
> Data:    Mer, 23 Maggio 2007, 5:52 pm
> A:       [log in to unmask]
> --------------------------------------------------------------------------
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear Paul, I read the Dev Cell you suggested me: it’s impressive and
> useful. I’ll see how I can put it into practice for my purposes of
> membrane tracking.
> Thank you.
> Andrea
>
>
> Andrea Manazza, MD PhD
> Dept. of Biomedical sciences and human oncology
> Molecular Oncology-CeRMS
> Via Santena 7, 10126 Torino
> Italia
> tel: +39-11-6336859
> fax: +39-11-6336887
>
>
>>
>>
>> -------------------------- Messaggio originale
>> ---------------------------
>> Oggetto: [SPAM] Re: leading edge tracking
>> Da:      "Paul Herzmark" <[log in to unmask]>
>> Data:    Gio, 5 Aprile 2007, 11:43 pm
>> A:       [log in to unmask]
>> --------------------------------------------------------------------------
>>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Andrea,
>>
>> I think this paper might be useful to you. They mapped outgrowing
>> pseudopods
>> in great detail. Perhaps you can use their methods
>> Dev Cell. <javascript:AL_get(this, 'jour', 'Dev Cell.');> 2005
>> Feb;8(2):215-27. Related
>> Articles,<http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?itool=pubmed_Abstract&db=pubmed&cmd=Display&dopt=pubmed_pubmed&from_uid=15691763>
>> Links <javascript:PopUpMenu2_Set(Menu15691763);>
>> *A local coupling model and compass parameter for eukaryotic
>> chemotaxis.*
>>
>> *Arrieumerlou
>> C*<http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Arrieumerlou+C%22%5BAuthor%5D>,
>> *Meyer
>> T*<http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Meyer+T%22%5BAuthor%5D>
>> .
>> --
>> Paul Herzmark
>> Specialist
>> [log in to unmask]
>>
>> Department of Molecular and Cell Biology
>> 479 Life Science Addition
>> University of California, Berkeley
>> Berkeley, CA  94720-3200
>> (510) 643-9603
>> (510) 643-9500 fax
>>
>> On 4/5/07, Andrea Manazza <[log in to unmask]> wrote:
>>>
>>> Search the CONFOCAL archive at
>>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>>
>>> Sorry for the previous, untitled, posting. I now repost it, correctly
>>> titled.
>>> Thank you.
>>> A
>>>
>>> Dear confocalists, I'd like to ask you for your opinion on a protocol
>>> for
>>> analysing membrane dynamics. I work with human lymphoma cell lines, and
>>> they change their shape and behaviour depending on an inducible RNAi
>>> machinery: when I turn it on, cells loose their transformed aspect
>>> (becoming round-shaped), they give up forming membrane protrusions and
>>> stop their spontaneous and anarchic movement. I made more than 50
>>> imaging
>>> experiments, in different conditions, with different stimuli and
>>> different
>>> cell lines: the data are strong, biochemically confirmed, and we're
>>> really
>>> confident. I have now to measure and quantify the change, during the
>>> time-lapse experiments. I did the imaging with my TCS SP2 confocal,
>>> using
>>> the "transmission" modality, further modified in order to minimize the
>>> laser energy ( I can give you the details off-line).
>>> I tried to track the membrane movements with Metamorph, but I failed
>>> using
>>> the automatic tracking. This is due to the fact that the cells display
>>> an
>>> adhesion point, around  which the whole membrane continuously rearrange
>>> its shape, every few seconds. Sometimes they also migrate for short
>>> distances. My aim is to measure the generation and movement of membrane
>>> protrusions, quantifying the displacement of the leading edge.
>>> I tracked manually, frame by frame, the leading edge of any single
>>> cell,
>>> with the "track objects" option of Metamorph, moving the research frame
>>> every time, updating the position and going on. The graphics are
>>> convincing, but –during a recent oral presentation- I've been fairly
>>> criticized: they told that, regardless to the fact that data are
>>> absolutely reliable, my tracking procedure seems a bit odd.
>>> Would you mind sending me suggestions and critics, before I send the
>>> paper?
>>> I may send you a few photos of the cell movements, so you can take a
>>> look
>>> at it, if you think it's useful.
>>> Thank you in advance for your help (I checked the archive, without any
>>> real benefit).
>>> Andrew
>>>
>>>
>>>
>>>
>>> Andrea Manazza, MD PhD
>>> Dept. of Biomedical sciences and human oncology
>>> Molecular Oncology-CeRMS
>>> Via Santena 7, 10126 Torino
>>> Italia
>>> tel: +39-11-6336859
>>> fax: +39-11-6336887
>>>
>>
>
>