An innovative high-tech laser scanning microscope makes using state-of-the-art equipment a breeze.

In an effort to work smarter, not harder, simplicity in use is the key to a recently introduced laser scanning microscope (LSM) from Carl Zeiss MicroImaging GmbH, Hamburg, Germany—the LSM 5 EXCITER.

The new microscope system eliminates the need for manual operation as all moving components, such as emission filter wheels, variable beamsplitters, and the pinhole and mechanical attenuators for each laser line, are computer-controlled.

The LSM 5 EXCITER’s fully motorized scanning module is based on three major principles:

• Short beam path combined with highly sensitive detectors close to the specimen for maximum efficiency.

• High quality filters and dichroics selected for optimum match to available laser lines ensure highest transmission.

• Exchangeable filter sliders and dichroic wheels providing flexibility for present and future applications.

Relatively soft materials, such as polymers in a non-contact procedure, are analyzed by the system with high measuring accuracy and resolution. The LSM 5 EXCITER provides image sizes of up to 2,048 x 2,048 pixels, 12-bit resolution per channel, high scan speeds, high-sensitivity detectors (photomultipliers), and short light paths from the specimen to the detector. Two options are available, StitchArt plus and Topography for LSM.

StitchArt plus—The LSM 5 EXCITER has a StitchArt plus option which expands microscopy in all directions, such as extra-long line profiles or image stack arrays extending more than 100x the size of the scan field. This can be automatically acquired, exactly assembled, and precisely quantified.

In addition, the StitchArt plus option integrates many automatic procedures and adjusting functions, including, among others, autofocus, automatic contrast and brightness optimization, automatic orientation of the scan field, and precise first order correction of the objective’s residual field curvature.

Topography for LSM—The optional Topography for LSM software package handles the extraction, processing, presentation, and measurement of height information of 3-D surface topographies and 2-D surface profiles. Together with the LSM 5 EXCITER configured for materials analysis, this software makes up a system for fast, non-contact optical microprofilometry.

A 3-D topography of a diamond-coated angle grinder blade was captured with LSM 5 EXCITER in the StitchArt plus option. Image: Carl Zeiss MicroImaging

The same topography software with the identical range of functions is available for materials applications of the LSM 510, Zeiss’s earlier version LSM. The software is interactive and fully integrated with the user interface. With its clearly arranged controls, it can be mastered easily and does not require deep software literacy.

The Zen of LSM

The software needed for the LSM 5 EXCITER—Zen—uses a combination of state-of-the-art technology and intuitive user interface and is designed for use on a 30-inch screen.

Zen has a Pro-Basic concept that ensures tool panels are never more complex than needed. For instance, in Basic Mode, the most common tools are displayed. It is up to the user to activate Pro Mode and add additional functionality.

In many cases, the microscope system and the monitor are physically far apart. Therefore, when loading the sample and adjusting the focus, it becomes very hard to read the settings on the computer screen. This problem is solved with Zen’s unique interface, which is scalable.

Also, setting up conventional confocal software to perform a given experiment can take an extended amount of time and must be repeated with each experiment. Zen’s adjustments must be done only once and can be restored with just two mouse clicks. Each experiment’s layout can be set up and saved in addition to being shared in a multi-user environment.

LSM 5 EXCITER is offered with polarization equipment in combination with their Axio Imager and Axio Observer microscopes. Special emphasis has been placed on the flexibility of the LSM 5 EXCITER. Zeiss has seven versions with one or two confocal channels and combinations of lasers in the blue and green spectrum and a choice of four microscopes available.

— Adria Nieswand