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STATUS: The TIRF scope is currently being worked on. If you would like to use it please email Jennifer (jennifer_waters@hms.harvard.edu). To report a problem, please fill in our online Problem Report.

1. The illumination problem: I was unable to figure out the strange illumination pattern that was discovered a few weeks ago. Nikon flew in one of their application scientists to try to figure it out, and she was unable to find the source of the problem either. They decided to send us a whole new TIRF illuminator. It arrived last week while I and literally everyone from Nikon was at the ASCB meeting. I'm still waiting for someone from Nikon to come in to install it (I bug them daily about it), but there are other things going on as well that have held it up...

2. The new lasers: We have purchased a new laser system for TIRF which has MUCH more power than the old laser system. We have some new single molecule imagers who need more power, but everyone will benefit from the new lasers. The lasers are being installed and aligned today and tomorrow.

3. The new microscope: Nikon introduced a new inverted microscope stand at the ASCB meeting, called the Ti, as well as a new TIRF illuminator (that only works with the Ti). This scope and illuminator looks like it will be much better than the old stand we have....for example, the Perfect Focus system is integrated into the stand and should be more stable, and the TIRF laser angle adjustment knobs are now motorized so we can automate changing between two wavelengths for two color TIRF imaging.

Right now there are only a few of these scopes in existance in the US - they are still in production in Japan. But Nikon has agreed to send one of them to the NIC so that we can try it out and so that Sam Reck-Peterson and I can validate it for them for single molecule imaging. The new Ti will be installed in place of the old TIRF scope this Friday. If all goes well it should be available for use next week, but since it is a new scope I will need to meet with you to show you how to use it. Nikon has agreed to leave it here for a few weeks, but I am hoping that (if we like it and I beg enough) they will leave it permanently.

I am sorry that the TIRF scope has been out of order for so long, but as you can see there has been a lot going on during this time. And I am hopeful that once things are settled, the scope you will be working on will be a much better one.

The all day timelapse I ran on 7/2 failed - the software locked up with a 'server busy' error. I ran debugging software and found that this was caused by interaction of the software with the motorized condenser on the TE2000E. I fixed that problem and am running more tests to make sure the software is now stable.

I believe all of the problems with the TIRF microscope are now fixed. The motorization problem was identified, and the motorization and perfect focus have worked well for me in tests. I also found and fixed a loose connection in the camera which I am hoping was the cause of the camera error in MetaMorph. I am currently running an all day timelapse to confirm that all is working.

The replacement filter cassette arrived yesterday, and Pam Angevine installed it today. Unfortunately this did not appear to solve all of the problems. There are still intermittent problems with the motorization on the microscope. Nikon plans to install another new microscope in the coming weeks. A date has not yet been set.

Another problem as been reported as well: two users have reported camera errors that required restarting MetaMorph to resolve. We are replacing the computer on the TIRF microscope in the next couple weeks (I am waiting for the new computer to arrive). I will address this problem when the new computer is installed.

I spoke to Nikon today and was told that the new filter cassette is still on backorder. They do not know when a new one will be available.

Nikon installed a new motorized microscope and Perfect Focus unit on May 11. The new microscope appears to have a faulty motorized filter cassette. This causes an occasional 'Filter Block' error to come up on the microscope control pad. The error causes the microscope and MetaMorph to temporarily freeze. After 10-20 seconds, MetaMorph usually unfreezes, allowing the user to proceed. There are ways to set up most peoples experiments that allow us to work around this problem. If you want to use the microscope before it is fixed (see below), please email me (jennifer_waters@hms.harvard.edu) and I will meet you at the microscope to help you set up your experiment in a way that will hopefully avoid the problem.

Nikon is going to replace the filter cassette. They are currently waiting for a new filter cassette to become available, estimated to be the week of June 18th.

I'm afraid we are having problems with the motorization of the new TIRF scope. Nikon is not sure what is causing the problem, and is going to start by replacing one of the parts. Unfortunately this part will not be available for 2 weeks. In the meantime, the scope is available for use but there may be intermittent problems. Please email me if you would like to use the scope and I will meet you there to get you started and help you work around the problem.