Hi,
I second Aseem's suggestion regarding SImFCS.
A user of our core facility is using FV1000 and the SimFCS software from the
Gratton lab, it appears to work quite well, as far as I know. The SimFCS
software can be installed as a fully-functional 1-month demo, so you can
test it yourself before making the final decision. One big advantage of the
RICS approach that I see is the ability to subtract slow movements, such as
sample or stage drift.
A while ago I was told by Olympus that point scanning was not going to work
for FCS on the FV1000 system, but I do not quite remember their technical
explanation. From my tests, I remember there was an issue with the first or
last datapoint being of incorrect value, due to the timing of the AOTF
"shutter", or synchronization of the AOTF with data acquisition. You could
try point-scanning of the fluorescent plastic slide from Chroma, the
fluctuations that you see should indicate the instrument noise/instability,
including the laser.
The number of datapoints in point-scanning mode on FV1000 was limited to
4096, i.e., the max. permitted line width. I was told that before a scan a
piece of code is sent to the control board(s), essentially reprogramming the
firmware, and this somehow imposes a limit on the number of datapoints.
You could do time-lapse point scan to acquire multiples of 4096 points,, but
then you have an unknown amount of time between the time frames.
Anyway, I think SimFCS is worth trying.
Stan Vitha
Microscopy and Imaging Center
Texas A&M University
On Sat, 20 Feb 2010 22:09:52 +0530, aseem mishra <[hidden email]> wrote:
>Hi Jean,
>
>You might want to have a look at Enrico Gratton's work and specially
>SimFCS wherein one can calculate diffusion coefficients from an image
>collected in a point-scan mode. I do not know if a blinking laser
>gives you the possibility of doing FCS (Do let me know if such a
>possibility exists. I would love to try it at my place.) We rather use
>a hardware correlator and a simple laser (diode/HeNe)
>
>Aseem
>
>
>On Fri, Feb 19, 2010 at 3:27 AM, Jean-Pierre CLAMME
><[hidden email]> wrote:
>>
>> Hi all,
>>
>> I'm thinking about modifying our fluoview 1000 to do FCS. The fluoview has
>> the possibility to do single point measurement. However as we can still
>> choose the pixel time in this mode, I was wondering if the laser beam always
>> stays at the same point of interest and continuously illuminates the point
>> or if it actually "blinks".
>>
>> If it doesn't "blink", I would think it could hinder the FCS measurement et
>> thus I'm wondering if there is an option to change this in the soft. ?
>>
>> Thanks,
>>
>> JP
>>
>>
>>
>>
>> - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
>> Jean-Pierre CLAMME, PhD
>> Senior Scientist
>> Nitto Denko Technical
>> 501 Via Del Monte
>> Oceanside, CA 92058
>> E-mail: [hidden email]
>> Phone: +760.435.7065
>>
>
>
>
>--
>Aseem Mishra
>Senior Research Assistant,
>Malaria Group,
>International Centre for Genetic Engineering and Biotechnology,
>New Delhi-110067
>INDIA