Michael Hooker Microscopy Facility

Michael Hooker Microscopy Facility (MHMF.ORG)

What Was Down!

2004

Aug 30 - The Green 561 nm laser: The laser has been delivered. Installation is scheduled for Thursday, September 2. If all goes well the system should be fully operational on September 3.
Aug 16 - Leica SP2 green laser: The new solid state 561 nm diode green laser is expected to be installed in the last week of August. All other laser lines (351 UV1, 364 UV2, 458, 488 blue, 514, 633 red nm) are running normally.
Aug 06 - LeicaSP2 green laser: A solid state 561 nm diode laser is being purchased to replace the burnt out 568 nm Krypton laser. Delivery time is 3 to 6 weeks on receipt of the purchase order by Leica.
Aug 02 - LeicaSP2 Krypton laser: The Krypton (568 nm) laser tube is burnt out. This burn out is premature, and is a most severe situation. Tube replacement is costly, slow (~4 weeks shipping time) and has a severely limited life time (< 1 year). An alternative 561 nm solid state laser, although more reliable (> 3 yrs), has a quoted shipping time of 4 to 6 weeks (possibly 8 weeks). This laser is being ordered. In the mean time please consider the Zeiss 510 confocal. An alternative source for the Krypton laser is being investigated, but should not be counted on.
If you do not require DAPI the Zeiss 510 is an alternative confocal, which can scan FITC, GFP, Rhodamine, Texas Red, CY5, and similar fluorophores. It does not work well with Alexa 594. Please contact Michael if you want to use this machine and have not received orientation.
July 29 - LeicaSP2: The system is running now. However, with the system running, it was obvious that the Krypton laser (568 nm, Texas Red, Rhodamine, CY3, etc.) was not functioning. It seems that an electrical transient on July 18 took out a few circuit boards, the x-axis scanner motor, the Krypton laser and scrambled the firmware in the microscope & scan head. All the other lasers (Argon, 488 nm, HeNe, 633 nm & Argon UV, 351 & 364 nm) are operational. Repairs on the Krypton laser will continue on August 2 when a new power supply will be delivered so that the state of the tube can be determined.
July 29 - LeicaSP2: UV Laser is working! DAPI, Heochts, Alexa 350, Fura-2 etc. scans can be carried out.
July 28 - LeicaSP2: Repairs continuing. Waiting for parts.

July 26 - Aug 6 (or later) Renovations to Rooms 6129, 6033a & 6123

Thurston Bowles rooms 6129, 6033a & 6123: Facility renovations will be carried out from July 26 to Aug 6. Resources will be unavailable due to Physical Plant/Contractor work. Limited access to microscopes is possible but not guaranteed. See table below for very rough estimates of resource availability. Please make way for Physical Plant/Contractor workers.

Booked times can not be guaranteed.

The Leica inverted (FRESNEL) and Nikon (HOOKE) upright widefield microscope are still in room 6129 and can be used when work is not going on. Typically this is mid afternoon and after hours.

Nikon TE2000 inverted fluorescence scope (MOPHEUS & WHEATSTON) will not be available July 26 to Aug 2. Possibly availability Aug 2 to 6 at another location.

Perkin Elmer spinning disk confocal will not be available for the duration of the renovations.

The Leica SP2 confocal (LAPLACE) and the Leica Laser Micro Dissection system and Zeiss 510 confocal (HERTZ) will not be available when their respective rooms are being renovated.

Image Processing work stations (MONARCH & MALUS) may be located in the corridor during regular work hours. YOUNG will be located on the 8th floor of the Neuroscience Research Building.

Xerox color printer (\\MINSKY\PHASER860DX-MHMF) is located in the corridor

Date

Thurston Bowles

Zeiss 510

Leica Inverted

Nikon Upright

SP2

LMD

Monarch

Young

Nikon inverted

Perkin Elmer

6129

6033a

6123

confocal

widefield

widefield

confocal

workstation

workstation

widefield

confocal

Tue

Aug 24

OK

OK

OK

M

OK

OK

NRB

++

na

Late Aug

OK

OK

OK

OK

OK

OK

NRB

OK

++

OK = available
na = not available
++ = probably available
M = available but green laser for Texas Red/Rhodamine/Alexa568/Alexa594/CY3 etc. is not functioning. Other laser lines are working including UV for DAPI/Alexa350 etc. A 561 nm solid state laser is on order to replace the Krypton 568 nm laser.
NRB = work hours only, 8th floor Neuroscience Research Building

July 22 - LeicaSP2: Scan head repairs will continue on July 28. Waiting for a diagnostic cable to arrive.
July 22 - LeicaSP2 UV Laser: The flow sensor is now working. The UV laser will be reintegrated into the confocal system once the scan head is operational.
July 19 - LeicaSP2 UV Laser: flow sensor part is on back order and not expected until late July.
July 19 - LeicaSP2 Confocal: The Leica is not starting due to a SCSI bus controller problem. The system is not usable. Leica service has been notified and expected on July 21. Until the nature of the problem is diagnosed an estimated repair date can not be given.
July 16 - LeicaSP2 UV Laser: Flow sensor will now be re-installed on Thursday July 22 (not the impossible date of Wednesday July 20 noted below)
July 14 - LeicaSP2 UV Laser: The UV laser has been tested and is functional. However the UV laser is not useable due to a faulty flow sensor. The flow sensor needs to be replaced before the UV laser can be put back into service. Coherent is shipping out the part and it will be installed on Wednesday July 20, and if all else goes well the UV Laser should be functional. In the mean time the confocal is still useable with the visible excitation wavelengths (458, 488, 514, 568, & 633 nm).
July 12 - LeicaSP2 UV Laser: A missing part has delayed the reinstallation of the UV laser. The UV laser will be tested on Wednesday, July 14. The confocal system will not be available for use on that day. Otherwise visible excitation confocal scanning can still be performed.
July 9 - LeicaSP2 UV Laser: The EnterpriseII UV laser is back in Chapel Hill and will be re-installed, aligned and tested on Monday July 12.
July 1 - LeicaSP2 UV Laser: The EnterpriseII UV laser has been checked out and will be shipped back to Chapel Hill. The unit is expected back about mid July.
June 22 - Leica SP2 aobs confocal: The coupling fiber for the Argon laser has been realigned. The maximum 488 nm power at the stage is restored to 3.5 mW
June 8 - Web server for http://microscopy.unc.edu http://mhmicroscopy.med.unc.edu http://physiology.med.unc.edu http://confocal.med.unc.edu http://bric.unc.edu ftp://mhmicroscopy.med.unc.edu The web server for these site lost connectivity to the Ethernet. The most likely cause is a glitch in the Ethernet card of the computer that serves these pages. The card will be replaced as a precaution.
June 3 - LeicaSP2 UV Laser: Power supply has been shipped back to Leica.
June 2 - LeicaSP2 UV Laser: head has been checked out, and is OK. Power supply is being sent out for appraisal.
May 21 -LeicaSP2 UV Laser: The UV laser is still in Germany for repair.
May 12 - Zeiss 510 Meta: Axiovert 200 stand firmware timing was tweaked, and may improve alignment of the tube lens after switching between lsm and visual mode. It is conceivable that this change may cure apparent focus jumps and microscope stand/lsm lockups.
May 3 - Leica SP2 AOBS confocal: The UV laser has been shipped back to Coherent for repair. The expected turn around time is 4 to 6 weeks. During this period imaging DAPI, Hoechst and other dyes excited by 351 nm or 361 nm will not be possible. Other dyes which are excited by visible wavelengths may be used for DNA labeling, for example To-Pro 3 (Molecular Probes), propidium iodide (Molecular Probes). Also the new DRAQ5 (Axxora) is useful for DNA labeling in live cells. Although it tends to wash out of fixed cells it is still good for DNA labeling of fixed cells.
April 29: Leica SP2 aobs confocal: The UV laser is not functioning. All other lasers are still useable. Leica service has been notified and well be coming out on May 3 to assess the UV laser. Status of the machine will be updated on the booking page for the LeicaSP2. In the mean time UV scanning (DAPI, Hoechst, Alexa 350 etc) is not possible.
April 9: Leica SP2: software upgraded to Leica LCS software version 2.5 build 1237a.
Mar 16: The colour (made in Canada) MicroPublisher camera is back on the Leica DMIRB inverted microscope. Also C-Imaging has been updated to version 5.2.1 on that computer and now gives color previews when viewing in focus mode.
Mar 16: On Leica inverted widefield microscope C-Imaging has been upgraded to version 5.2.1- Nikon DXM1200 color camera will now white balance.
Zeiss 510 confocal: The Axioscope will be sent out for service due to corrosion on the turret mirror. This mirror requires replacement, which can only be done at the factory since it must be critically aligned. Confocal will be unavailable for 7-9 weeks at a date yet to be decided.
Zeiss 510 confocal: A spot is appearing on the confocally scanned image. This is particularly noticeable on transmitted light views but also darkens fluorescent views.

Feb 10: Leica DMIRB inverted microscope: The color MicroPublisher camera on the inverted Leica fluorescence & DIC widefield scope is damaged. This camera is being sent back to the factory for repair.
- Images can still be captures with the B/W camera (Hamamatsu OrcaER).
- Multi-channel/color fluorescence can be captured by switching filters manually (see Jan or Michael for more information).
- Color photomicrographs e.g. H&E or PABS can not be taken. Use the Nikon Microphot instead
Jan 28: Leica SP2 confocal: Kypton laser and power supply have been replaced. All excitation lines are available.
Jan 23: Leica SP2 confocal: A power supply for the Krypton Laser has been ordered. It will be installed some time between Jan 27-30.
Jan 21: Leica SP2 confocal: The Krypton laser is not functioning. It is not possible to perform confocal excitation at 568 nm (green), so Texas Red, CY3, Rhodamine and similar fluorophore scanning is not possible. However scanning with the other lasers (Argon 458, 488, 514 nm & HeNe 633 nm) is still possible. Leica service will be called. Note that the Krypton laser may be burnt out and hence require replacing. A replacement may take a few weeks to arrive. Leica service engineer is visiting on Friday Jan 23 (Service Job #660409). Status updates will be posted here.
Jan 14, 2004: Zeiss 510: The transmitted light condenser has been repaired. Time outs should not occur and simultaneous transmitted light and confocal scanning can be done again.

2003

Dec 12: Zeiss 510: Transmitted light condenser aperture drive is not functioning. Condenser aperture is stuck in the open position (high numerical aperture). Zeiss LSM software will report KAB1 errors, which can be ignored. Software condenser aperture control also does not work. This breakdown limits adjusting contrast while viewing by eye in transmitted light and DIC mode. Also effected is laser scanned transmitted or DIC viewing. Zeiss service has been informed, ref # 213098.
Dec 8: PE Spinning disk: reflection issue is resolved. Imaging is once again working.
Nov 19: Leica SP2: software upgraded to Leica LCS software version 2.5 build 1227.
Nov 18: Spinning Disk Confocal PE Ultraview: Strange reflections are occurring when 568 nm or 647 nm laser excitation is selected. Perkin Elmer have seen this problem. Scan head may be shipped out for service.
Nov 14: Q-Imaging color CCD camera: The Q-Imaging color CCD camera may have a bad fire wire cable, which causes a message saying that it can not communicate to the computer to appear. The cable has been replaced. If any problems crop up while using this camera please contact Michael so that further action can be taken.
Nov 10: Microphot upright microscope & DXM 1200 color camera: have both been cleaned. The microscope view is much cleaner and permanent spots on the camera have been removed.
Nov 5: DXM 1200 Color camera on Nikon upright/dissecting scopes: SimplePCI version 5.2 on the computer called Hooke will not color balance the Nikon DXM 1200 camera. The work around for now is to use SimplePCI version 5.1. Icons for running version 5.1 are on the desktop and on the start menu.
Oct 31: Spinning disk confocal UltraView: shading/light leak in TexasRed/Rhodamine & CY5 bands. Service confirmation no. 310389966. Open Temporal mode, default image control box, blue line, go live, then raw to live without doing setup get DCAM error. Also DCAM error when setup binning to non square size. Always do square.
Oct 29: Perkin Elmer Spinning disk confocal: serial interface system upgraded, emission filters upgraded, software upgrade to version 5.2.27 & OrcaER bios upgrade.
Oct 17: Perkin Elmer Spinning disk confocal: serial cable stretched, plug and socket damaged due to the computer table being moved. Serial interface box may be damaged. Control to shutters and filter wheel not functioning reliably. Do not rearrange tables etc. Damage due to user abuse is not covered under maintenance contracts and repair cost will be charged to users.
Oct 9: Leica SP2: Krypton laser mirrors realigned. Improved output measured at the tube to 22 mW from 12 mW. Reduced noise from this laser. The laser seems to be slightly noisy still at 9 o'clock power knob setting. To avoid this noise go to the 10 o'clock position on the power knob.
Oct 8: LeicaSP2: 568 nm channel is sometimes noisy. Could be noisy laser, or AOTF or fiber mis-alignment. Service will be performed on Oct 9.
Oct 7 & 8: Zeiss meta: AOTF calibration was linearised and emission filter wheel to PMT #2 was repaired.
Sept 26: Zeiss 510 meta: Emission filter wheel before PMT #2 is still sticking. Sticking is less of a problem when wheel is progressed from top to bottom in the pull down menu. Zeiss service has been notified.
Sept 15: Zeiss 510 meta. The laser module has been realigned which has improved the power available from the argon laser (458, 477, 488 & 514 nm) at the microscope stage by an order of magnitude. The emission filter wheel to PMT #2 has been adjusted so that it will now position correctly.
Sept LeicaSP2: miscellaneous problems: 1. Display mode may need to be set to dual monitor mode. 2. Scan settings may need to be set to 1.0x (UV). 3. Toggle off auto zoom mode after choosing zoom region.
July 23: Bulb for the Fiber Optic Light used on the Nikon Dissecting Scope: The bulb for the Schott Fiber Optic lamp is on back order. The lamp will be installed as soon as it is received.
July 16: Leica SP2 confocal. The first hard disk drive has been replaced. This drive maps to the C: & D: drive letters in Windows. Leica Confocal Software (LCS) user profiles have been restored as much as practical. However the organization of the Leica Software has changed slightly for some users, so differences, including missing programs, may be noted. If you have any problems please contact Michael.
July 10: Leica SP2 confocal. Is running again. The disk drive finally booted up to WindowsNT 4. Some disk errors were encountered during a low level SCSI verification. Leica will replace the drive on July 16. The drive replacement will
July 10: Leica SP2 confocal. The confocal is currently not available. The operating systems is not booting from the hard drive. A service call has been placed with Leica, and further information will be posted here at is becomes available.
June 17: Leica SP2. X-galvo replaced & upgraded.
May 22: Perkin Elmer spinning disk confocal: Occasionally camera errors are being returned when running the temporal mode software. A software and hardware upgrade is scheduled for June. Camera issue will be addressed during upgrade.
May 20: Phasor 860 solid dye color printer: Printer has repaired. Heater was blown out and the print head stop operating correctly.
Apr 24: Zeiss 510 meta confocal: is working. Software has been upgraded to version 3.2. There are some minor changes. e.g. must set to visual mode manually after LSM scanning.
Mar. 05: Perkin Elmer confocal. The OrcaER CCD camera shows some apparent blooming on top left of the image. It is worse with binning, and more noticeable with higher camera gain. There is also a claim that there are ~7 bad pixels on the chip. This is not good and has mentioned to Perkin Elmer service.
Feb. 26: Leica4D confocal computers are no longer be connected to the Medical School Network:
The Leica4D confocal (Viceroy) is no longer connected to the Medical School Network, and hence the entire internet, including the Cytic_Fibrosis domain. Scanning can still be performed and the 100 MByte Zip drive can be used to transfer data. OIS has discontinued the sub-network that the 4D was assigned to on Feb. 18, 2003 as a final act in a 5 year plan to give the 152.2. subnet back to the main campus, and block that subnet in the Medical School. Unfortunately the 4D OS9-VME computer can not be easily be migrated to the Medical School subnet. Hence the 4D is isolated from the network. Alternative solutions are being investigated, but are limited by the potential fragility of the two unsupported operating systems that are an essential part of the 4D confocal.
May 20: Phasor 860 solid dye color printer: Printer has broken down. It will be unavailable until repair is completed. Xerox were notified on May 20. No estimated repair time is available yet.
Apr 22: Zeiss 510 confocal: Due to a software glitch the Zeiss LSM software is not able to scan. Zeiss service is scheduled to commence Friday morning (04-25). System status will be posted here on Friday and booked users will be notified by e-mail.
Apr 10: Zeiss 510 confocal: The laser light output from the 458 nm line of the Argon Laser is not being controlled correctly by the AOTF. At 0% there is 458 light output with remains at the same level until ~20% above which it decreases until ~45%. Above 45% light output increases slightly. Zeiss service has been contacted. Work order 199609.

During April 15 to 17: LeicaSP2 confocal & Leica AS-Laser Micro-Dissection. Air conditioning improvements were carried out in Room 6123 Thurston Bowles. This work is required to keep the room temperature under control when the confocal lasers are running. The Leica Laser Micro Dissection system will be temporarily moved into room 6129.
Apr 6: Zeiss 510 confocal. Laser safety shutter checked out. May be light getting through
Mar 25: Zeiss 510 confocal. The shutter to the binocular head is sticking shut.
- Symptom - can not see any light when viewing through the oculars with transmitted light or fluorescence.
- The work around:-
  - In LSM 510 software set Microscope Control to viewing mode and turn on transmitted light.
  - Use a 3 mm Allen wrench (yellow arrow points to wrench which is inserted in loosening screw) and loosen the binocular head while supporting it.
  - (click on image to zoom in)
  - Remove binocular head.
  - In front of the tube lens is a black shutter which will swing out of the way when gently nudged.
  - (long arrow points to shutter. short arrow points to clamping screw)
  - Replace binocular tube and tighten screw
  - While supporting tube double check that the binocular head is firmly clamped on
- Note that this shutter is part of the laser safety system which prevents direct viewing of laser light beam. Do not do anything to jam this shutter open permanently.
- Repair order has been called in. Due to a number of Zeiss repair engineers unavoidably being out of the country, repair will not be carried out until April.

Mar. 18: Leica SP2aobs. The overheating Krypton laser has been replaced. The UV AOTF modulator has been replaced.
Mar 13: Nikon Upright Microscope (MicrophotSA). The disk drive has been replaced. Images stored on the drive is currently unavailable. The Act-1 software, which runs the DXM 1200 color camera has been re-installed. Note that Norton Antivirus prevent writing of valid CDs if real time file protection is on.
Mar. 11: LeicaSP2aobs. The Krypton laser (568 nm, Rhodamine, Texas Red) is overheating. The power knob needs to set below 9 o'clock, and preferably at the minimum knob power level to get laser light at the stage.
Mar. 11: Nikon Upright Microscope (MicrophotSA). The computer which runs the camera and C-Imaging has been infected by a Trojan back door virus. The operating system has been wiped clean and essential security updates are waiting for ATN/OIS to remove the network access restrictions to this machine. When full access is enabled the complete operating system can be restored and C-Imaging installed
Mar. 10: Nikon Upright Microscope (MicrophotSA). The computer for this system called \\Hooke has been taken off the network. This machine's software is infected. Please carefully transfer only image files off this computer using the CD writer.
Mar. 07: LeicaSP2aobs. The Krypton laser has been repaired. All lasers on the system are fully function.
Feb. 27: LeicaSP2 confocal Krypton laser, 568 nm (Texas Red) is unusable due to mechanical failure. Leica service has been notified and will be making a service visit Monday, Mar. 3 or Mar. 6 or 7.
Feb. 18: Leica SP2 Krypton laser replaced.
Feb. 10, 2003: LeicaSP2 Krypton laser has failed. A new laser has been ordered. Installation date has not been set yet.
Feb. 7, 2003: Zeiss Argon laser has been replaced. Laser modules have been realigned.
Jan. 31, 2003: Zeiss 510 confocal's Argon laser is failing. Power output is declining and at higher currents it is cutting out. Replacement is tentatively set for Friday 02-07. Booked users will be requested to yield their time to the Zeiss service. In the mean time please avoid running the Argon/2 laser at Output (%) greater than 50% and ensure that the tube current is under 7.0 amps. If the tube blinks out switch to standby, set output (%) to minimum, wait about a minute and switch laser to on. Now output(%) may be increased.
Jan. 2003: Leica LMD is running off the network.
Jan. 2003: LeicaSP2 Confocal AOBS - new computer is installed. When switching to scan check that dichroic has turned to a blank position and the arc lamp light path is closed.

2002

Nov 19. - Perkin Elmer Ultraview spinning disk conofcal - Software non longer detects/runs camera and shutter after upgrade to version 5.2. Perkin Elmer will be back on Nov 25 to start to complete repares.
2002 September: Meta detector is need of alignment on the Zeiss 510 meta. A service request has been made. ref. no. 188749. The Meta detector is still useable, however the sensitivity is not as high as it should be.
2002-08-29: Leica SP2 confocal is unavailable for acquisition since the upright microscope stand's mechanical focus system is running erratically, and is apt to smash slides. Analysis may be performed on the machine, but the microscope control box should not be turned on.
2002 May: Hamamatsu B/W C5985 cooled camera on the Leica inverted fluorescent microscope (computer: Fresnel formerly Huygens) is not functional. An OrcaER cooled B/W camera is currently installed, and will be replaced with a DVC1312. Image acquisition with all the cameras on this setup may be done using C-Imaging
2002 June: Acquisition using the Q-Imaging Micropublisher cooled color camera through Photoshop on Fresnel is not available - This function may be restored in the future. In the mean time images may be acquired using C-Imaging or Q-Imaging


		Copyright 2001-08 Dr. M. Chua, Office of Research, University of North Carolina, Chapel Hill, NC 27599
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Last Updated: 2007-09-08