Z-drift

[Nathalia Holtzman]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi all,
I need some help. We have a 1 year old Leica SP5 inverted scope and pretty
much all of a sudden we started to get z-drift. Every time you do a time
lapse, no matter what the z-step or time interval, the system does not start
at the top of the sample, it moves down significantly and progressively.
Anything I can do to fix this?

I have called the service rep but would like to fix the problem ASAP. It is
possible that I am the first person to use the system for this application
since we upgraded the software and firmware.

Nathalia

--
Nathalia Glickman Holtzman Ph.D.
Department of Biology SB D328
Queens College, CUNY
65-30 Kissena Boulevard
Flushing, NY 11367

Office: 718-997-3678
Lab: 718-997-3517
Fax: [phone]

[Ray Hicks-3]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi Nathalia,

I'm not sure if it's the same problem, but we have had a similar problem
twice on the DM IRBE inverted body of our SP1, it turned out to be immersion
oil that had dripped down the objectives and into the mechanism causing the
turret to sink slowly - the loss of focus was visible over a short period of
time using the eyepieces.

Our field service engineer fixed it quickly both times (the first time was
approximately a year after we got the scope, the second time was much much
later, due mainly to better cleaning and care from the users once they were
made aware of the risk).


Cheers

Ray



----- Original Message -----
From: "Nathalia Holtzman" <[hidden email]>
To: <[hidden email]>
Sent: Thursday, November 08, 2007 1:58 PM
Subject: Z-drift

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi all,
> I need some help. We have a 1 year old Leica SP5 inverted scope and pretty
> much all of a sudden we started to get z-drift. Every time you do a time
> lapse, no matter what the z-step or time interval, the system does not
> start
> at the top of the sample, it moves down significantly and progressively.
> Anything I can do to fix this?
>
> I have called the service rep but would like to fix the problem ASAP. It
> is
> possible that I am the first person to use the system for this application
> since we upgraded the software and firmware.
>
> Nathalia
>
> --
> Nathalia Glickman Holtzman Ph.D.
> Department of Biology SB D328
> Queens College, CUNY
> 65-30 Kissena Boulevard
> Flushing, NY 11367
>
> Office: 718-997-3678
> Lab: 718-997-3517
> Fax: [phone]

[Andrew Resnick]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Nathalia,

Not knowing anything about your setup or what you are looking at, my
first guess would be temperature fluctuations.  Try and let the
microscope and stage warm up for a couple of hours, enclose as much
as possible in some sort of  chamber (a large bag may work in a pinch).

One check- during z-drift, does the z-encoder indicate that the
z-position is fixed? Another check- try setting upper and lower
limits for a z-stack and (rapidly) move between those tow
setpoints.  Is there hysteresis present?  Those can tell you if there
is a hardware problem.

Andy

At 08:58 AM 11/8/2007, you wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hi all,
>I need some help. We have a 1 year old Leica SP5 inverted scope and pretty
>much all of a sudden we started to get z-drift. Every time you do a time
>lapse, no matter what the z-step or time interval, the system does not start
>at the top of the sample, it moves down significantly and progressively.
>Anything I can do to fix this?
>
>I have called the service rep but would like to fix the problem ASAP. It is
>possible that I am the first person to use the system for this application
>since we upgraded the software and firmware.
>
>Nathalia
>
>--
>Nathalia Glickman Holtzman Ph.D.
>Department of Biology SB D328
>Queens College, CUNY
>65-30 Kissena Boulevard
>Flushing, NY 11367
>
>Office: 718-997-3678
>Lab: 718-997-3517
>Fax: [phone]

Andrew Resnick, Ph. D.
Instructor
Department of Physiology and Biophysics
Case Western Reserve University
216-368-6899 (V)
216-368-4223 (F)

[Jon Mulholland]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Nathalia,
         We have been experiencing the same thing on our SP2 AOBS
system.  This not a simple "drift" problem but a large sudden drop
downward which only happens when attempting to use the time lapse
setup.  Like you we are seeing this after a software and firmware
upgrade.  We are still trying to identify the problem but suspect it
is the software/firmware itself as it only happens using the
time-lapse setup.  We'll let you know what we discover, we have a
very good Leica service engineer.  Please let us know if you learn anything.
Thanks
Jon

At 05:58 AM 11/8/2007, you wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hi all,
>I need some help. We have a 1 year old Leica SP5 inverted scope and pretty
>much all of a sudden we started to get z-drift. Every time you do a time
>lapse, no matter what the z-step or time interval, the system does not start
>at the top of the sample, it moves down significantly and progressively.
>Anything I can do to fix this?
>
>I have called the service rep but would like to fix the problem ASAP. It is
>possible that I am the first person to use the system for this application
>since we upgraded the software and firmware.
>
>Nathalia
>
>--
>Nathalia Glickman Holtzman Ph.D.
>Department of Biology SB D328
>Queens College, CUNY
>65-30 Kissena Boulevard
>Flushing, NY 11367
>
>Office: 718-997-3678
>Lab: 718-997-3517
>Fax: [phone]

Jon Mulholland, Director
Cell Sciences Imaging Facility
Beckman Center, B050
Stanford University School of Medicine
Stanford, CA 94305-5301

voice 650-725-7532
fax    [phone]

http://taltos.stanford.edu 

[Jacqueline Ross]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Nathalia,

We had a problem like this on our SP2 some years ago. This may not be that relevant because we have an upright scope with a galvo stage but it turned out that the galvo actually needed recalibrating. We had a Leica engineer recalibrate it and it has been fine since then.

Cheers,

Jacqui

Jacqueline Ross

Biomedical Imaging Microscopist
Biomedical Imaging Research Unit 
School of Medical Sciences 
Faculty of Medical & Health Sciences
The University of Auckland
Private Bag 92019
Auckland, NEW ZEALAND

Tel: [phone] Ext 87438
Fax: [phone]

http://www.health.auckland.ac.nz/biru/ 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Nathalia Holtzman
Sent: 09 November 2007 02:59
To: [hidden email]
Subject: Z-drift

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi all,
I need some help. We have a 1 year old Leica SP5 inverted scope and pretty
much all of a sudden we started to get z-drift. Every time you do a time
lapse, no matter what the z-step or time interval, the system does not start
at the top of the sample, it moves down significantly and progressively.
Anything I can do to fix this?

I have called the service rep but would like to fix the problem ASAP. It is
possible that I am the first person to use the system for this application
since we upgraded the software and firmware.

Nathalia

--
Nathalia Glickman Holtzman Ph.D.
Department of Biology SB D328
Queens College, CUNY
65-30 Kissena Boulevard
Flushing, NY 11367

Office: 718-997-3678
Lab: 718-997-3517
Fax: [phone]

[Stephen Cody]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Nathalia,

I would guess that 95% of the time, z-drift is caused by temperature
instability in your room, incubator or both, rather than a mechanical
fault with your microscope. This has been discussed many times
previously on this list. You should search the archives for "z-drift" or
"Focus Drift". Objective lenses are made of brass which will expand and
contract with very slight changes of temperature. It is not a trivial
matter to control the temperature of the room and incubator accurately
enough to be able to perform time lapse microscopy. I have just spent
two weeks playing with the air-conditioning system and incubator to
achieve stability after the hospital switch from winter heating to
summer cooling.

To gain more understanding of your problem while you are waiting for a
service call I suggest you look at the tutorial I have placed online at:

www.ludwig.edu.au/confocal/drift

If you follow this procedure you should be able to work out if the
movement of the lens is always in:
-A downward direction - Gravity causing drift may be the cause

-An upward direction - Your lens is still warming up to the incubator
temperature

- Oscillating up and down
        - temperature instability caused by heating or cooling
        system in the room.
        -Or a microscope incubator system with either an on/off
        controller (get a controller with "fuzzy logic" or "PID
control")
        -PID controller requires "Tuning" (read the manual).

Temperature control is not trivial, you need to invest quite a bit of
time to get it right.

Cheers


Stephen H. Cody
Microscopy Manager
Central Resource for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008 Royal Melbourne Hospital
Victoria,      3050
Australia
Tel: [phone]    Fax: [phone]
email: [hidden email]
www.ludwig.edu.au/labs/confocal.html
www.ludwig.edu.au/confocal

Tip: Learn how to receive reminders about you microscope booking:
www.ludwig.edu.au/confocal/Local/Booking_Hint.htm  

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Nathalia Holtzman
Sent: Friday, 9 November 2007 12:59 AM
To: [hidden email]
Subject: Z-drift

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi all,
I need some help. We have a 1 year old Leica SP5 inverted scope and
pretty
much all of a sudden we started to get z-drift. Every time you do a time
lapse, no matter what the z-step or time interval, the system does not
start
at the top of the sample, it moves down significantly and progressively.
Anything I can do to fix this?

I have called the service rep but would like to fix the problem ASAP. It
is
possible that I am the first person to use the system for this
application
since we upgraded the software and firmware.

Nathalia

--
Nathalia Glickman Holtzman Ph.D.
Department of Biology SB D328
Queens College, CUNY
65-30 Kissena Boulevard
Flushing, NY 11367

Office: 718-997-3678
Lab: 718-997-3517
Fax: [phone]


This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research does not waiver any rights if you have received this communication in error.
The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research.

[Amanda M. Lawrence]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi Dr. Cody,
I tried to follow the links to your tutorial suggested in this email.  The "home page" comes up but when I click on any of the menus at the top of the page (tutorials, image gallery, etc.), I get the message "this page cannot be found".

As I am still new to confocal microscopy I am interested in your tutorials esp. the one on time-lapse and controlling z-drift, since time lapse is one thing our facility is going to have need of shortly.

Thanks so much,
Amanda Lawrence
Electron Microscope Center
Mississippi State University

>>> Stephen Cody <[hidden email]> 11/8/2007 6:45 PM >>>
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal 

Dear Nathalia,

I would guess that 95% of the time, z-drift is caused by temperature
instability in your room, incubator or both, rather than a mechanical
fault with your microscope. This has been discussed many times
previously on this list. You should search the archives for "z-drift" or
"Focus Drift". Objective lenses are made of brass which will expand and
contract with very slight changes of temperature. It is not a trivial
matter to control the temperature of the room and incubator accurately
enough to be able to perform time lapse microscopy. I have just spent
two weeks playing with the air-conditioning system and incubator to
achieve stability after the hospital switch from winter heating to
summer cooling.

To gain more understanding of your problem while you are waiting for a
service call I suggest you look at the tutorial I have placed online at:

www.ludwig.edu.au/confocal/drift

If you follow this procedure you should be able to work out if the
movement of the lens is always in:
-A downward direction - Gravity causing drift may be the cause

-An upward direction - Your lens is still warming up to the incubator
temperature

- Oscillating up and down
        - temperature instability caused by heating or cooling
        system in the room.
        -Or a microscope incubator system with either an on/off
        controller (get a controller with "fuzzy logic" or "PID
control")
        -PID controller requires "Tuning" (read the manual).

Temperature control is not trivial, you need to invest quite a bit of
time to get it right.

Cheers


Stephen H. Cody
Microscopy Manager
Central Resource for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008 Royal Melbourne Hospital
Victoria,      3050
Australia
Tel: [phone]    Fax: [phone]
email: [hidden email]
www.ludwig.edu.au/labs/confocal.html
www.ludwig.edu.au/confocal

Tip: Learn how to receive reminders about you microscope booking:
www.ludwig.edu.au/confocal/Local/Booking_Hint.htm  

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Nathalia Holtzman
Sent: Friday, 9 November 2007 12:59 AM
To: [hidden email]
Subject: Z-drift

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal 

Hi all,
I need some help. We have a 1 year old Leica SP5 inverted scope and
pretty
much all of a sudden we started to get z-drift. Every time you do a time
lapse, no matter what the z-step or time interval, the system does not
start
at the top of the sample, it moves down significantly and progressively.
Anything I can do to fix this?

I have called the service rep but would like to fix the problem ASAP. It
is
possible that I am the first person to use the system for this
application
since we upgraded the software and firmware.

Nathalia

--
Nathalia Glickman Holtzman Ph.D.
Department of Biology SB D328
Queens College, CUNY
65-30 Kissena Boulevard
Flushing, NY 11367

Office: 718-997-3678
Lab: 718-997-3517
Fax: [phone]


This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research does not waiver any rights if you have received this communication in error.
The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research.