Bugs

This page is a not-necessarily-complete record of pending and past technical problems with the LMCF microscopes.

6/30/08 - SP5 - Dana fixed the UV collimation optic wheel - replacement part proved unnecessary as adjusting the tension on the motor solved. System working much better, pinhole alignment good. Stage seems ok still. Laser powers -

405 = 4.3 mW
458 = 1.5 mW
476 = 2.7 mW
488 = 7.0 mW
496 = 1.75 mW
514 = 5.3 mW
561 = 6.0 mW
594 = 0.52 mW
633 = 3.4 mW

6/16/08 - SP5 - continued problems with the stage. Problem with UV beam expander, dana realigned all but still not great. Power at objectives reasonable but still down from install -

405 = 3.98 mW
488 = 7.0 mW
561 = 5.5 mW
594 = 0.45 mW
633 = 3.06 mW

Dana will send manual 3-plate stage until motorized stage problem is resolved. Other issues with scan head - AOTF vis sweeps were flat, pinhole alignment was bad. Swapping scope control box seems to be helping with the stage. UV wheel got worse.

4/16/08 - SP5 - installed insulation on power supply. Dana aligned lasers and measured power while there (to turn off aotf blanking - LAS diag/testcenter/scanner low level saotfm 0, zoom >x25)-

405 = 4.9 mW
458 = 1.75 mW
476 = 3.0 mW
488 = 7.8 mW
496 = 1.94 mW
514 = 5.8 mW
561 = 6.1 mW
594 = 0.52 mW
633 = 3.3 mW

1/18/08 - SD2 - measured laser power (out of laser, before fiber), run at full power after 5 min warmup, 488 line = 31 mW, 568 line = 33 mW. In comparison, kiehart wounding scope produced ~40 mW after fiber = all lamda.

11/28/07 - SP5 - replaced AOTF controler for visible lines, all visible lines then controlled properly. Measured laser powers and they seemed to be as install (the argon laser didn't warm up for very long): 12/3/07 installed AOTF controller for 405 and power measured as 5.8 mW. 561 measured as 6.3 mW. It then died, new one installed 12/10/07, powers measured as 405 = 5.2 mW, 561 = 6.3 mW.

488 = 8.1mW
514 =6.5 mW
561 = 6.5 mW
594 = 0.55 mW
633 = 3.6 mW

11/13/07 - SP5 - FRAP bleaching not that great on checking laser powers AOTF seem to be either on or off and offer no AT control. Diag gives okay frequency, powers about 2x over and install. Laser powers measured:

405 = 4.83 mW
488 = 6.81 mW
561 = 5.0 mW
594 = 0.16 mW
633 = 2.33 mW

9/18/07 - 510 inverted - On opening the software a stream of CAN_ error messages appeared (lack of communication with the laser modules). Appeared to be a problem where the PC was unable to open port7000 to communicate to the real time computer. Spent time looking for problems in firewall or TCP/IP settings. Problem was failure of the main data board downstream of the realtime computer which for some reason makes the RT computer unable to connect to the main PC.

9/5/07 - SP5 - Installation. All went well, except scope had additional port on side not compatable with the incubator, scratched shield on the condensor, no DAPI cube,

Laser powers through 10x objective, lasers at full power, aotf 100%, deactivate x blanking, 400 Hz, zoom 32

405 = 3.9 mW
458 = 1.6 mW
476 = 2.9 mW
488 = 8.4mW
496 = 2.2 mW
514 = 7.2 mW
561 = 6.0 mW
594 = 0.53 mW
633 = 3.6 mW

8/22/07 - SD1 - Laser power low. Measured power at 10x objective with BN meter and Kiehart lab meter before and after mirror and fiber alignment. Before alignment: BN (K)
488 = 130 microwatt (87 microwatt)
568 = 141 microwatt (62 microwatt)
647 not visible or detectable

After alignement:
488 = 342 microwatt (220 microwatt)
568 = 537 microwatt (230 microwatt)
647 = 390 microwatt (125 microwatt)

5/07/07 - Live cell station - Focus drift from faulty encoder in docking station, charlie disabled it until new part arrives. New one seems okay.

April 2007 - New Stereoscope - Broken dial, Middle TL dial doesn't stay forward and sometimes sticks depending on the position of the others. Need - HE red filters, dual light guide, 1x c-mount. Holding down RL and TL minus buttons = double tap. Custom buttons for all users and check reliability.

3/13/07 - 510 - Todd noticed when tiling, one edge of field consistantly brighter than the other, possibly not completely flat also. Charlie fixed with scan field rotation macro.

3/7/07 - Axio - Metamorph crashes after DIC acquisition, shutter closes but camera fails? Check hard ware settings and see if fault is persistant.

11/22/06 - 410 - PMT 1 was replaced with a brand new Hamammatsu cooled PMT. Contrast values still register some 100 points higher than pre-PMT upgrade, but pictures & sensitivity look great. Scanners aligned and shading adjusted on PMTs. External lasers aligned. Microscope cleaned & cover relabelled. All this service was performed by the senior service technician (Brad Bobbitt).

11/17/06 - 510 - Alex measured laser power. In Admin mode 7.5 amp 80/20 10x = 1.2 to 1.4 mW. 3.67 mW with 488/543 dichroic. Seemed some problem in measure in normal mode. Alex recalibrated AOTF to improve linearity power, went up to about 1.3 mW. Not certain if this equates to increased bleaching or just measuring? Scanning must be in bidirectional mode to measure, uni = half. Need to remeasure with Dan's power meter to check is off comparable sensitivity.

11/13/06 - SD - A few people complain of having worse crosstalk problems than before. Really need to upgrade with an emission filterwheel. Problem might be because the scope is just more sensitive than before?

10/31/06 - SD - Couple of users and have had problem with laser igniting.

10/19/06 - SD - Mike Smith installed new fiber and aligned laser scanhead etc. Replacing 568 filter removed image imperfections in that channel. 647 filter was beginning to show signs of burning so Mike is going to post me a replacement. Whole system working well. Measured laser power (warmed up for about 10 min) - 50 mW total: 488=8.3 mW, 568=8.5 mW, 647 13.1 mW. Guidline for when replaced under SC - <5mW per channel. Cleaning cost ¥300 000 ($2500).

10/12/06 - SD - Installed cleaned scanhead. Looks much better but still some imperfections visible on the red channel but much reduced. New fiber yet to be installed. Also thoroughly cleaned the microscope and surroundings, installed 63x/1.4 oil objective from BN scope. Returned scanhead had no details of what they had done.

10/5/06 - 510 - Further improvement of laser power by Alex and Charlie. Laser power at objective 4 fold higher: New fiber (transmits 75%, old one 45%), improved alignment of fiber into scanhead, primary VIS mirror and routing mirror - better centered on back aperture of objective (moving collimator leads to movement of 3 pixels). No obvious damage to scanning mirrors but y mirror had a slight hum. Cleaned dust from primary mirror. Board controlling Ph3 broke, replacing at 3:00 10/6/06. Done.

Laser power measured at 10x objective, 80/20 objective (power at install) all 2-3 higher with optimal dichroic

* 351 nm (0.785 mW)
* 364 nm (2.02 mW)
* 458 nm 0.24 mW (0.391 mW)
* 477 nm 0.38 mW
* 488 nm 1.4 mW (2.19 mW) same as demo 1.2-1.4 mW "good"
* 514 nm 0.804 mW
* 543 nm 0.090 mW (0.126 mW)
* 633 nm 0.36 mW (0.451 mW)

9/25/06 - 510 - laser power too low for rapid bleaching. Back aperture of objectives dirty, primary mirror of Ar/2 adjusted = 25% improvement but the laser power measured at objective at least 5 fold too weak. Lasers themselves not adjusted but all working well -
543 - 1.12 mW; >1.00 after AOTF
633 - 4.92 mW out of laser; 4.67 mW after AOTF
Ar/2 @ 7.5 amps - total after first mirror 35 mW (all lines), after AOTF

* 458 3.20 mW
* 477 4.26 mW
* 488 16.0 mW
* 514 8.79 mW

(ie >32 of 35 mW reaches fiber optic to go to scope)

laser output delivered through 10x objective is pathetic - 488 0.4 mW, want more than 2.0 mW, red lasers similarly bad. Possibly the fiber optic - but illumination looked even when objective removed (couldn't take out fiber because of UV/Vis alignment). If not fiber, primary dichroic could be blemished = factory repair in germany. Alex and Charlie checking further next week.

9/12/06 - Axio - Upgrade: Uniblitz shutter fine, MAC 5000 controller for LUDL filterwheel can't be driven by metamorph through USB so we need to get extra I/O card for pc. Thread pitch of 25 mm analyzer different to filters/filterwheel, need to get unthreaded version, homemade one for now. Bill installled threadless analyzer 10/5/06. Uniblitz shutter i have doesn't fit on ex filterwheels, needs an adapted form from shutter - about $950 so keep using ND wheel for now. 10 for 6 position?

9/06 - 510 - Ar/2 laser getting weak, pinhole alignment fine, Alex checking laser at end of month.

8/7/06 - ? - Spinning disk - Away for cleaning, internal mirrors needed cleaning and perkin elmer had to send it back to yokogawa in japan. About 3 weeks has turned into about 8 weeks.

7/06 - Axio - Audrey noticed that DIC images differ between have illumination set to none and any setting and having RL shutter closed. Not sure why, possible software having to move filterwheel or not affects exposure time even though not in light path?

7/13/06 - Axio - Various problems with metamorph, required resetting of all electronic equipment, hot pixels remained from camera controller problem? Temporary problem.

7/11/06 - 510 - stage had become loose, Alex replaced missing screw and tightend. Fine now, problem was only noticable if stage was far forward.

6/29/06 - 510 - Find function produced unusable gain values for the transmitted light detector. Maintain/setfind/AutoB+C controls. Select ChD Two sliders default values = 100, set amp gain to 1.0 checked.

6/29/06 - 410 - PMT2 high contrast/sensitivity problem proven to be in the detector itself (moved to detector position 1, and the high contrast values moved with it). Another PMT off of LSM Tech's demo unit was overnighted and installed in detector position 1, replacing the malfunctioning PMT. While this replacement PMT has contrast values some 150 points better than our old one, it isn't brand new and is ~50 contrast points less sensitive than our detector currently occupying position 2.

NEEDED: LSM Technologies needs to supply us with a *brand new* detector and to switch the PMT currently occupying position 2 back to position 1. Ultimately, we should have the more sensitive PMT in detector position 1.

6/23/2006 - 410 - failure of PMT3. No image, no noise. Dave Myers replaced failed socket for PMT3, now functional.

6/22/2006 - Axio - Halogen bulb not working, output switch on back had been changed.

2006 - Axio - DIC quality. Since changing microscope stands the DIC quality is sometimes poor. Installing new 100x seems to improve. Bill Faught will bring HC slider - currently testing.

5/24/2006 - Axio - Unevenness of Hg illumination. Despite realignment and bulb change seems to be a tendency for a bright central stripe. Aligning the bulb using the 100x objective improves as this is where the problem is worst.

5/15/2006 - Axio Imager - MetaMorph crashed upon opening, no obvious cause, changing to single user mode then back to multi user mode solved the problem.

5/13/2006 - 510 - Transmitted light appeared not to be functional, user had entered previous image mode so the scope was offline. Other issues with transmitted light - Alex replaced cable, errors still occur.

5/2006- 410 - Several problems with PMT2. Filters were incorrectly installed, after changing these contrast values still seem high. The pinhole was replaced for PMT2 in optic upgrade but is working normally and not the cause of the problem. Brad improved brightness of samples under Hg lamp - cleaned collecting lens of Hg arc lamp.

5/11/2006 - Spinning Disk - PM visit from Mike Smith, Laser alignment improved power increased to 50 mW at scan head. Fiber might be slightly imperfect - some haze around bright central spot. 60/1.4 seems ruined by oil, 60/water added. Dark spots around edge of image - contamination in scan head as rotation of camera see the contamination patterns rotate with the image; rotation of the entire scan head and camera sees the contamination pattern fixed and the image rotates. Green filter cube has red transmission.