http://confocal-microscopy-list.275.s1.nabble.com/Uniformity-of-2-photon-illumination-tp6719119p6726443.html
Just set your power control up as close to your laser as possible. I use a
for most imaging applications 200mW or less is plenty. I also keep a few
movable beam blocks handy at all times. If I need to work with the laser, I
throw multiple blocks into the system, then remove them as I go. That way
the laser is never going further than the next block in the system. Here is
table, while the other I keep up close. The magnetic holder allows you to
easily move and reposition them. By using multiple blocks, I ensure there
is always at least one somewhere in the beam. I only remove the last one
when I am ready to shoot the beam into the microscope. The only caution I
out of the beam rather than lifting them straight up. (you can paint the
ability to move the block up and down. Collar R2 can help cover up the post
> Hi Stuart, in short, I align the 2P beam just as described by Julio in
> previous e-mail reply. I suggest you call your Zeiss service rep and then
> watch how he aligns the 2P to the vis laser(s). Once you've seen it, the
> protocol should be very easy to replicate.
> You may already have a saved configuration called "2P Align" in
> C:AIM/settings that you can use. Also, you should have received a mirror
> slide with a grid on it that is perfect for this kind of alignment.
>
> Adjusting the beam path as Julio described is quite easy to do.
> Also, don't look into the laser light with your remaining good eye (first
> heard this sound advice from George McNamara).
>
> Cheers,
>
> Brian D Armstrong PhD
> Assistant Research Professor
> Light Microscopy Core
> Beckman Research Institute
> City of Hope
> Dept of Neuroscience
> 1450 E Duarte Rd
> Duarte, CA 91010
> 626-256-4673 x62872
>
>
>
http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:
[hidden email]]
> On Behalf Of stu_the_flat
> Sent: Thursday, August 25, 2011 3:38 AM
> To:
[hidden email]
> Subject: Re: Uniformity of 2-photon illumination?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> *****
>
> Thank you very much for your replies.
>
>
>
> Just to clear up a few points, I always
> take Z sections of the slide just to make sure it wasn’t a case that the
> slide
> was at an angle, (The system is not designed to hold slides so its always a
> bit
> “Heath Robison” trying to get a slide underneath it!)
>
>
>
> I had checked various sites around the
> slide and I was getting consistent results, yesterday I scanned the slide
> with
> the descanned detectors and got a similar image.
>
>
>
> We want all the power we can get as would
> love to be able to image more deeply into cardiac muscle, (is it a fair
> assumption that more power will allow us to image deeper?) for that reason
> we
> have a Coherent Chameleon Ultra II, and I believe its peak power is around
> 5.5W. However would the alignment not take place “down stream” of the OPO?
> So I
> could just set that to its minimum intensity setting.
>
>
>
> Anyway it is kind of irrelevant because the
> system is under service contract with Zeiss, So all I need to do it get on
> the
> phone and start whining at them, Although I would love to give laser
> alignment a
> go myself!
>
>
>
> Once again thank you for all your replies
>
>
>
> Yours
>
> Stuart McIntyre
>
>
>
> --
> View this message in context:
>
http://confocal-microscopy-list.588098.n2.nabble.com/Uniformity-of-2-photon-illumination-tp6719119p6723940.html> Sent from the Confocal Microscopy List mailing list archive at Nabble.com.
>
>
> ---------------------------------------------------------------------
> *SECURITY/CONFIDENTIALITY WARNING:
> This message and any attachments are intended solely for the individual or
> entity to which they are addressed. This communication may contain
> information that is privileged, confidential, or exempt from disclosure
> under applicable law (e.g., personal health information, research data,
> financial information). Because this e-mail has been sent without
> encryption, individuals other than the intended recipient may be able to
> view the information, forward it to others or tamper with the information
> without the knowledge or consent of the sender. If you are not the intended
> recipient, or the employee or person responsible for delivering the message
> to the intended recipient, any dissemination, distribution or copying of the
> communication is strictly prohibited. If you received the communication in
> error, please notify the sender immediately by replying to this message and
> deleting the message and any accompanying files from your system. If, due to
> the security risks, you do not wish to receive further communications via
> e-mail, please reply to this message and inform the sender that you do not
> wish to receive further e-mail from the sender.
>
> ---------------------------------------------------------------------
>
>