Laser Microdisection Systems: Past, Present and Future

Modern devices, like the Leica Laser Microdisection System, make it a lot easier to decipher the different cell types within a sample. Yet, it was once a lot harder. It took years of science and industry to create the advanced systems researchers use today.

Leica Laser Microdisection System

Before Laser Microdisection Systems

The quality of research wasn’t as great.  The reason? Samples like eukaryotic tissue or ecological samples contain numerous different cells. For researchers to see a clear picture of the problems they intend to identify, they needed to isolate samples down to more granular levels.

In the time before laser microdisection systems, researchers often had to look at samples as a whole. This approach gave a good description of the general tissue makeup, but failed to identify underlying compositions and details.

 

Solving the Problem

The solution came with cancer research. Researchers looking at cells wanted a way to dissect small portions of tissue. The portions of tissue could then be tested separately and their unique compositions discovered.

Lance Liotta of the National Cancer Institute noted his team’s laser microdisection solution in a 1996 science paper. The method involved a transparent film placed over a sample tissue section. The specific cells would adhere to the film. An infrared laser would also aid in the removal of the sample portion. With the sample separated, the researchers could test it on its own.

 

Modern Laser Microdisection Systems

The use of microdisection systems have spread beyond cancer research.  Researchers use them for numerous areas: live cell research, climate research and cover-slip engraving for electron microscopy.  Modern systems can quickly isolate cells, have convenient laser manipulation and allow researchers to easily mark and track microscopic samples. Unique lasing within fluorescence is also available in Leica Laser Microdisection Systems.

 

The Future of Microdisection

There’s no telling for sure what will change when it comes to laser microdisection in the future, but chances are the process will become increasingly simpler for researchers, allowing them to advance mankind’s collective understanding of microscopic structures. To learn more about one of the most advanced systems available, check out the Leica LMD7000 and see exactly how far laser microdisection systems have progressed.

Microscopes in a Teaching Environment

Recently, Leica Microsystems Product Manager Vince Vaccarelli gave his insights into what makes a good microscope for teaching. While having the most up to date technology is nice, these are some factors you should consider when purchasing microscopes for the classroom.

  1. Portability. Classroom microscopes should be small and easy to handle especially when you consider the amount of times you’ll be taking them (and their respective cables) in and out of storage.
  2. Sturdiness. Being exposed to rough handling and various environmental conditions means that durability is key for both universities and schools.
  3. Usability. Ideally, classroom microscopes should have well-labled parts, few adjustable features, and components that can’t be removed. Students should be able to easily work with these microscopes.
  4. Optical performance. Sacrificing good optics would be defeating the purpose bringing microscopes into the classroom. With good optical performance you get better contrast, color, and resolution that reproduces images accurately. By Giving your students a clear and crisp image you enable them to properly learn about what they’re seeing.
  5. Maintenance. After a busy day of learning, servicing a fleet of microscopes can be daunting. Keep in mind the difficulty of cleaning and maintaining the microscopes when considering your next purchase.

Leica has an educational product line with features such handles and cord wraps , mold growth resistance, and an anti-bacterial additive. All this within compact bodies for easy storage.

Correcting Aberration in Stereo Microscopy

Stereo Microscopes are optimized for use with samples exposed directly to air. Even though this creates increased resolving power, there are drawbacks once the sample is embedded in polymer of immersed in liquid.

This is due to refractive index mismatch which creates aberration.

Aberration occurs when there is a difference in the refractive indices of air and water. This makes it difficult to accurately observe certain features. Fortunately, this can be remedied with a specialized objective or a correction collar that corrects the mismatch. With greatly reduced spherical aberration, a sharper image is produced.

Abberation 2

Leica’s solution to aberration comes in the form of microscopes that use a correction collar. With a correction collar you can make adjustments to a group of lenses within the objective and correct for refractive index mismatch. This makes the image plane smaller giving the sample a sharper, crisper focus.

Whether the subject is embedded in a polymer (e.g. glass, plastic, etc.) or immersed in liquid we have what you need to handle a variety of obstacles to get the best image out of your sample.

Holes and Depressions: Look Deeper with Shadow-Free Illumination

Born out of necessity from surgical microscopy, near vertical illumination allows for observation into deep bored holes and recesses.

Now you don’t have to resort to surgical microscopes to benefit from this feature. The Leica LED5000 NVI and LED3000 NVI are attachments that integrate two powerful LEDs along the optical path. Anything from cartridge cases, to cylinder heads, and injectors can be viewed with shadow-free illumination.

Leica LED5000 NVI

This is a modular accessory that fits on all high end stereo microscopes. Even with the small distance between the sample and the objective, the Leica LED5000 NVI still offers a shadow-free result. If you have an M-series stereo microscope then you’ll want to go with the Leica LED3000 NVI.

“Users preferring a different color temperature can put any commercially available filter into the integrated filter insert,” said Matthias Schacht, product manager. “This is useful for generating the light characteristics of a halogen light source, for instance.”

This is also useful for reducing reflections when looking at metals or other shiny surfaces.

Shadow-free illumination is useful in those very specific situations, but it’s better to be prepared for any kind of sample.

Don’t be left in the dark.

A New Way to Teach in the Classroom: Wi-Fi Capable Microscopes

Leica is introducing two new options to streamline classroom sharing, teaching and usability.

The first is the new Leica EZ4 W educational stereo microscope with a built-in Wi-Fi-capable 5-megapixel camera.

LMS_EZ4W

The second is the Leica ICC50 W Wi-Fi-capable 5-megapixel camera add-on that has been built to fit between the viewing tube and the body of any manual Leica DM compound microscope.

ICC50-W

These instruments are a useful resource when paired up with the free Leica AirLab App available on both iOS and Android. You’ll be able to capture, annotate, share, and organize your images.

“Since Learning content is transferred directly to the students’ devices, teachers can engage them more easily and promote team work. They can share results, work together, and network wherever they are,” said Vince Vaccarelli, Leica Microsystems product manager. “The interactive, digital platform increases attention and simplifies note-taking as well as assignments with image annotations.”

The standalone Leica EZ4 W microscope comes with:

  • 8x to 35x magnification
  • Zoom ratio of 4.4:1
  • 7-way LED illumination

If you already have an upright compound microscope then the Leica ICC50 W add-on might be a better fit for you.

Microscopy in the classroom just got a lot more accessible.

The Next Push Forward in Digital Microscopy: The DVM6

If you’re looking for some new horsepower to put behind your quality assurance/control, forensic, or failure analysis the newest iteration in the DVM series is here.

The DVM6 streamlines the digital microscopy process. It has been crafted with ease of use in mind so don’t let the size intimidate you.Leica DVM6 side

Swapping out objectives and tilting the microscope can all be done with one hand while it stays in focus. Also, PlanApo corrected lenses along with the 10 megapixel camera and convenient lighting options provide a crisp, clean image on screen. All of this at more than 30 frames per second.

Thanks to the encoding on the DVM6 your results are reproducible. The illumination, position, magnification, etc. are saved for every image since all of the instrument components are sensor controlled.

Leica DVM6 user

This works in tandem with the LAS X software. With it you can use Live Image with High Dynamic Range to instantly see every detail. There are options to create single shots, stitch together larger ones, measure in 2D and 3D, and annotate. LAS X helps any user create reliable, accurate data.

Here is the feature list for the DVM6 straight from Leica:

  • Manual or motorized versions
  • All system components encoded – also for the manual version
  • Motorized versions are hybrid and can be operated manually as well for fast coarse positioning
  • Zoom module with 16:1 zoom range
  • Integrated 10-megapixel high-resolution camera
  • PlanApo-corrected Leica optics with long working distance
  • Motorized and software-controlled Iris diaphragm
  • Integrated ring light and coaxial LED illumination
  • Snap-on adapters for ring light contrasting (polarizer, diffusor, low angle illumination)
  • Backlight illumination for translucent samples
  • Tilting stand for one-handed operation, tilting from -60° to +60°
  • Focus drive with a travel range of 60 mm
  • XY stage with a travel range of 70 mm x 50 mm
  • Autofocus with two options: one shot on region of interest, or continuous autofocus
  • LAS X software

A Small Lesson in Multiphoton Microscopes

Maybe this is an excuse to show off these incredible images created with the Leica TCS SP8 MP, but why not take a look back at where this technology came from while we’re at it?

Screenshot-47

What we’re seeing here is a 3D image with optical sectioning using a multiphoton microscope. The phenomenon behind this technology is called two-photon absorption (TPA). Essentially, two photons are absorbed at the exact same time in order to excite a molecule from one state to a higher energy electronic state. Thinking back to high school chemistry, light is given off once that molecule decays back to a lower state.

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Maria Goeppert-Mayer, the German-born American theoretical physicist, first predicted this process in her doctoral dissertation in 1931. It wasn’t until 30 years later, with the invention of the laser, that experimental verification for TPA was possible.

TPA was used as a spectroscopic tool until the 1980s. Once more developments in the field occurred, different applications were demonstrated. A few of them being photodynamic therapy, optical data storage, and imaging (obviously).

Screenshot-50

Watt W. Webb is best known for suggesting using TPA for imaging and microscopy. In 1990 he co-invented mulitphoton microscopy along with Winfried Denk and Jim Strickler. Earlier in his career, Webb pioneered techniques in fluorescent correlation spectroscopy (FCS). The combination of TPA and FCS resulted in high resolution, high signal-to-noise images.

One of the biggest advantages to multiphoton microscopes is the use of long wavelength, low energy excitation lasers. This is less damaging to live cells and introduces fewer toxic effects. This unique attribute is responsible for the in vivo microscopy you’re seeing in these images. Developments in medical endoscopy are being explored since the potential for in vivo, in situ real-time diagnostics is there.

This concludes our small lesson in multiphoton microscopes. Class dismissed.

High Resolution uPAINT Localization Technique with the Leica SR GSD 3D

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via: Leica Science Lab

Imaging living cells is always a challenge for most of the common super-resolution principles. Unlike the STORM and PALM methods, universal Point Accumulation for Imaging in Nanoscale Topography, or uPAINT offers super-resolved images and single molecule trajectories at very high densities with the Leica SR GSD 3D
Continue reading High Resolution uPAINT Localization Technique with the Leica SR GSD 3D

Explore the Hidden Worlds All Around Us

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If you’re reading this blog, chances are you’re no stranger to the ways of microscopy, but you can’t help but be amazed by the stunning textures and landscapes it can reveal.

Our latest example comes by way of the artist Pyanek and the Amazing Worlds Within Our World project. With subjects ranging from kitchen sponges to cornflakes, to soap bubbles, Pyanek takes us on a fascinating dive into more mundane objects of everyday live to explore their inner complexity.
Continue reading Explore the Hidden Worlds All Around Us

Best Microscopy Images of 2014

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2014 // 3rd // Dr. Igor Siwanowicz // HHMI Janelia Research Campus // Ashburn, VA, USA

We can all agree nature has some pretty amazing spectacles to show us at every scale. As our technology improves we get to bare witness to these displays at ever smaller scales.

In pursuit of these unseen mysteries, the worlds of art and science collide each year in a glorious display of technical prowess and curiosity at the BioScapes International Digital Imaging Competition.

This year’s big winner was video of a developing fruit fly embryo made from 30-second snippets pieced together from the first 24 hours of a fly’s life. The video is a fascinating view of cells multiplying and differentiating as the larva goes from a blob to a developed creature and begins to crawl away.

Check out the video below and check out all the top winners at Wired Magazine.