Light Microscopy Service & Support

Since its acquisition in 2011 TILL Photonics, now FEI Munich GmbH  has become an integral part of FEI Life Sciences. TILL Photonics' 20 years of experience in developing light microscopy systems has resulted in a unique technology portfolio which wasrecently successfully applied to standard setting correlative light and electron microscopy (CLEM) solutions, solutions which are greatly received by our customers. We will further build on this success and increase efficiency and speed in developingand deploying new correlative solutions to the market. This requires focusing our efforts.

Therefore, from January 31, 2014 onwards, all our light microscopy technologies and techniques will be based on the CorrSight platform in order to offer an even broader range of innovative cutting-edge correlative solutions. All technology and know-howof the light microscopy systems will continue to be used and will be further developed in our CLEM offerings. The non-CorrSight based stand-alone product lines iMIC and its options (iMIC Andromeda, TIRF, 42, and 2P), Intravital and the Calcium Imagingsystems will be discontinued.

For service requests please contact LM.Service@fei.com or call +1 866 547 8455 (US) or +49 89 895 662 0 (rest of world).

Important Notice:

As of April 1, 2015, systems which were produced until 2004 will be transistioned to an end-of-support status.

  Download letter

Repair process (RMA)

In order to return your equipment for repair, please request a RMA number. Shipment instructions will accompany the RMA number.

RMA Procedure
Health & Safety Declaration-en
Health & Safety Declaration-de

Live Acquisition Software

Please contact FEI Light Microscopy Service (LM.Service@fei.com) for Live Acquisition download link.

 

Hardware/Software FAQs

 

Hardware Support

 

Image Acquisition

 
  Q: I use TILLvisION on two computers. One of them is mainly used for data acquisition while the other is used for analysis. In the analysis mode I always have to wait while the program is searching for (non existing!) hardware. Is there a way around this?
A:  You can disable the Imago driver to skip the hardware initialization. Use the "Tools/Device Manager" menu and uncheck the "TILL IMAGO" entry.
 
  Q: I cannot figure out how to use the "GET AOI" button to select a region for image acquisition within a protocol. It works fine for me when I use it from the GRAB MENU (take a big image, draw an AOI over a portion of it and then GET AOI - results in new sizes and offsets for subsequent snapshots) but I cannot get it to work for me within a protocol following the instructions in the manual. I take a snapshot, draw an AOI, open the protocol window (now the snapshot is not the active window anymore) click get AOI - nothing happens.
  A: THE ORDER of steps is crucial!

Make sure that you

 
  1.  
  2. start the ProtocolEditor,
  3.  
  4. select the image with AOI in Vision,
  5.  
  6. (task-)switch to ProtocolEditor (either by pressing Alt-Tab or by clicking on the task bar on the bottom of the screen,
  7.  
  8. hit GetAOI.
  9.  
 

More technical:

 

TILLvisION and the TillProtocolEditor are separate programs for technical reasons. They communicate via COM (Component Object Model)-Interface, formerly known as OLE. If you press Get AOI, the Protocol Editor asks Vision for its active window and the first AOI. If there are many images but none of them active, this request fails.

 

So, after starting the ProtocolEditor and BEFORE hitting Get AOI, you have to make sure that the image window with the respective AOI is activated in Vision.

 
 
  Q:  Can I perform averaging of image information while acquiring images?
A:  Yes, there is an average and an accumulate mode which are mainly meant for video cameras used in conjunction with an 8-bit frame grabber. With our digital IMAGO camera you probably prefer to integrate on the chip by simply increasing the exposure time. You will get a much better signal-to-noise ratio by doing so. Only if integration on the chip fills the pixels beyond their full well capacity, digital averaging may yield a further increase in signal-to-noise ratio.
 

QE Camera

 
  Q:  How do I enable the high-gain mode of the Imago QE camera?
  A: >To enable the gain mode (Low light mode) of the Imago QE camera in TILLvisION 4.0.x or earlier, you have to open the
 

"tillpitp.ini" file.

 

The "tillpitp.ini"-file is located in the "c:\programs\tillvision\program"-folder.

 

HINT: the drive letter might differ from system to system as will the first folder "programs" be different on different language versions of windows.

 

To enable the gain mode you might have to add a line in the

 

    [default]
  section of the INI-file.
  In case that there is no line with
  highgain=0
  , add one.
 
 

 

To enable the high-gain mode you have to replace the "0" with "2" (without the quotes).

 
 

Sensicam / Imago grabber

 
  Q: What Sensicam/Imago grabber do I have?
 
 
  A: There are two versions of the Sensicam/Imago grabber card. The older one is called "520 KP", the new one "525 KP".
 

  How the two grabbers can be distinguished?

 
  •  
  • The OLD grabber is longer than the new one.
  •  
  • The NEW one has a red and a green LED on the grabbers front panel
  •  
 

  The older gabber card will most likely not work in modern computers (manufactured after 2001).

 

Symptoms can be:

 
  •  
  • The computer will not boot at all
  •  
  • The computer will boot after the reset button of the computer was pressed
  •  
  • You can only record one Image then TILLvisION will not take anymore images
  •  
  • The card is not recognized, resulting in an error -3 or -4 on TILLvisION start up
  •  
 
 
grabber_old_and_new_465
 
 
 

Software Support

 

Image Acquisition

 
  Q: I use TILLvisION on two computers. One of them is mainly used for data acquisition while the other is used for analysis. In the analysis mode I always have to wait while the program is searching for (non existing!) hardware. Is there a way around this?
A:  You can disable the Imago driver to skip the hardware initialization. Use the "Tools/Device Manager" menu and uncheck the "TILL IMAGO" entry.
 

High speed acquisition

 
  Q: How fast can I get? What is the maximum frames per second I can record?
A:  The maximum frame rate depends on several factors like resolution, binning and the camera readout speed and mode. Please read on to find out more. You will find out more as we give you an insight on the technical background in image aquisition.

  After the acquisition of an image the camera sends it at a rate of 25MB/second to the frame grabber board. That corresponds to a rate of 12.5MHz for the pixel transfer rate (16 bit information). The PCI frame grabber has two memory banks, each equipped with 8MB RAM. From the frame grabber the image data is transferred to the computer RAM via the PCI bus.

Since the frame grabber has two banks of memory it is possible that the camera writes data into the second bank while simultaneously the computer reads the images from the first bank. It is obvious that if you acquire more than 8 MB of data, the system has to switch between banks during the measurement.

  The TILL Protocol Editor uses the hardware in the following way: All images within one repeat loop are grabbed into one bank. Since the images of the next repeat loop are grabbed into the other bank, the system has to switch between the banks. This switching and its implications need a few milliseconds, depending on the chosen image size.

  As a result of this, the highest acquisition rate is achieved if you grab as much data as possible into one bank to avoid bank-switching. We call this "bursting acquisition" in contrast to the continuous acquisition mode where you do not care about the bank switching.
 

Camera & display settings

 
  Q: How can I control the camera and display settings?
A: To acquire live images (i.e. grab images and display them on screen), simply press the Live button. You will get images which are acquired and displayed according to the chosen settings. The program uses the same image buffer for consecutively acquired images and only keeps the last acquired image after the stop.

If you acquire an image of the type Fluorescence, the monochromator switches to the appropriate wavelength at the beginning of the Camera Exposure Time and switches back to the dark position at the end.

 To acquire only one image you can use the Snapshot button.

  Image Type:

 
  •  
  • Transmission - the monochromator stays in the dark position
  •  
  • Infrared generates a TTL signal synchronized with the image exposure
  •  
  • Fluorescence switches to the chosen wavelength during exposure
  •  
  Binning: is a synonym of on-chip-integration. With a binning factor of two, the camera accumulates four adjacent pixels to improve both sensitivity and speed, at the expense of resolution. As a result of lowered resolution further image processing is faster because of smaller image size.
 

Imago QE camera

 
  Q: How do I enable the high-gain mode of the Imago QE camera?
  A: >To enable the gain mode (Low light mode) of the Imago QE camera in TILLvisION 4.0.x or earlier, you have to open the
 

"tillpitp.ini" file.

 

The "tillpitp.ini"-file is located in the "c:\programs\tillvision\program"-folder.

 

HINT: the drive letter might differ from system to system as will the first folder "programs" be different on different language versions of windows.

 

To enable the gain mode you might have to add a line in the

 

    [default]
  section of the INI-file.
  In case that there is no line with
  highgain=0
  , add one.
 
 

 

To enable the high-gain mode you have to replace the "0" with "2" (without the quotes).

 
 

Protocols

 
  Q: How do I set up a wavelength protocol?
A:  For setting up an aquisition Protocol you will need the TILL Protocol Editor. The TILL Protocol Editor is an additional program which enables you to set up much more flexible protocols than with the Grab Settings/Image Series (Which is also not supported any more!). You can run the TILL Protocol Editor from TILLvisION or as an independent program.

The structure of the protocols which are arranged in the so-called Experiment Plan is very similar to the tree structure of the Windows NT Explorer. To edit the protocol you can use the Experiment menu or the appropriate Append and Child buttons. Full Drag & Drop and Copy & Paste support is provided too.
 
  Q: How to start and benchmark a protocol?
A:  The experiment setup is stored in a so-called Till Protocol. You can save it as single file and / or embed it into a TILLvisION Workspace (*.vws; visION document).To execute an embedded protocol you simply have to select it in the workspace view.

  Experiment Check Phase

Before executing a protocol, the Imaging System has to check if your system meets the experiment requirements like hard disk space, hard disk and memory speed, etc. The system performance may vary due to memory segmentation and parallel running processes.

While benchmarking the system, the TILL Protocol Editor first checks if the acquisition timing is "slow" enough to write directly to hard disk (about 2-3 MB/sec). If not, TILLvisION has to allocate physical memory exclusively for storing images during acquisition. You can approximate your physical memory needs by calculating the size of all images to acquire high speed and continuity. Add approx. 24 MB for Windows NT and the TILLvisION application.

The benchmarking is done on a per image sequence base, so it is possible to set up a protocol with high speed images at experiment start (written to memory) followed by less fast image acquistion (written to disk) and high speed acquisition at the end (again kept in memory).

  Experiment Acquisition Phase

If your data acquisition is not too fast for user interaction, you can mark events while acquiring data. This may be useful if you want to know later at what time in relation to image acquisition you have applied your specific chemical.

At the end of the experiment TILLvisION writes all images to disk to free physical memory for further experiments. The offline handling of experiment data makes use of the unique Big Image Driver, which transparently handles images on disk as if they were in memory. This feature allows the fast handling of images or image sequences much bigger than physical memory. Normally the location of images (memory or disk) is specified automatically.
 
  Q: How do I control loops and arrange images in a protocol?
A:  The Repeat page lets you set the number of repeats and the Cycle time. The cycle time is the time from the beginning of a cycle to the beginning of the following one. The End of Loop Delay occurs only once per repeat loop. The Accumulated Command Duration is the time which will be needed for the whole repeat loop and the Memory Consumption shows the necessary memory for the whole repeat loop (Size of Image * Number of Images).

  Used Grabber Bank Memory: Here you can see how much memory is used for one repeat-loop. All images from one repeat-loop will be stored in one frame grabber bank. Since one bank contains 8MB of memory, this value has to be less than or equal to 8MB. Acquiring images into one grabber bank enables overlapped exposure (acquiring of an image while the other one is read out).

  Arrange Images as: Due to the fact that increasing the number of images in one grabber bank may be useful, this option enables you to resort the images on the fly (more about high speed acquisition).Repeat command options
 
  Q: How do I modify the camera settings in a protocol?
A:  The Camera Settings Command allows to specify general camera options, namely binning, skipping and chip window size. Due to the fact that imaging is a very memory-consuming business you are enabled to reduce the image resolution to fit your needs. Furthermore you may (and should) reduce the camera readout size if you are only interested in a part of the whole image.

  Binning is a synonym of on-chip integration. With a binning factor of two the camera accumulates four adjacent pixels to improve both sensitivity and speed, at the expense of resolution.

  Skipping: Due to the properties of a CCD-Chip the complete charges of the CCD-Chip have to be removed from the chip before acquiring a new image. That means even if you acquire only an AOI, i.e. a smaller portion of the whole chip, the charges of the whole chip have to be removed. Since you are not interested in some regions of the chip it is not necessary to convert the charges to digital values (this means skipping the data). Therefore it is possible to skip them with a higher binning factor. Thus, you can gain acquisition speed by using fewer rows.
 
  Q: How do I do online analysis in a protocol?
A: The Protocol Editor offers you a wide range of Online Analysis options. The number of live images and live kinetics is only limited by your desktop size.

To activate online analysis, you have to insert an Online Analysis command behind an Acquire command. To get a live ratio, you obviously have to define two image acquisitions to calculate the ratio from. To see the image(s) during the acquisition, select Image. If you like to get an online kinetic, activate Kinetic in the Display dialog box. Then additionally you have to select an image which continues the previously defined ROIs (Regions Of Interest).
 

Export of image or kinetic data

 
  Q: I opened a sequence for export as an AVI file but when I tried to save the file I got an error message like "The selected filter is not able to save this object". What is wrong?
A: When you tried to save/export the sequence you selected the view of a frame of the sequence. Vision interprets this as if you wanted to save that special frame of the sequence as a single image which is incompatible with the AVI format. Switch to the workspace and select the sequence you want to export. That should solve the problem.