Confocal Fluorescence Microscopy Lasers

Lasers and Confocal Fluorescence Microscopy

Confocal Microscopy Laser Source Requirements:

•  Wavelengths: Most popular are 405nm, 488nm, 532nm/561nm, and 638nm. Others are often used as well. Wavelength combiners are ideal to cover all wavelengths.

•  Output power: 50 – 500mW with good stability

•  Beam Requirements: TEM₀₀ & low noise. Usually 1x PM fiber-coupled.

Multi-Wavelength Combiners: A Critical Tool for Fluorescence Imaging

CW multi-wavelength combiners have proven to be a critical component in Life Science applications, reducing time, cost, and complexity, and fundamentally changing these applications, allowing for increased efficiency and ease of use. Given the immense degree of complexity and time involved with combining multiple laser beams into a confocal microscope, multi-wavelength combiner modules have flooded the market in recent years. These plug-n-play style modules allow for swappable laser modules and quick and easy coupling with a confocal microscope, saving hours in precision alignment efforts. Laser combiner modules also help minimize the complexity of selecting multiple different wavelength sources from various laser models and even from various manufacturers, likely with different integration requirements. With a huge selection of wavelength combiners on the market, picking the right solution for your needs can be a difficult task. We provide a highly flexible, customizable series of multi-wavelength combiners, allowing for up to 7 excitation wavelengths of your choice in one user-friendly, compact unit.

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Multi-Wavelength Laser Sources for Multi-Color Fluorescence Microscopy

Otto Heimstaedt and Heinrich Lehmann, where the first to realize the possibility of fluorescence microscopy while experimenting with ultraviolet microscopes between 1911 and 1913. While their initial intention was to increase the spatial resolution of the microscope by using shorter wavelength light, they quickly realized that ultraviolet excitation was inducing autofluorescence in their samples [4]. However, these first instruments suffered from one major drawback, the high energy ultraviolet light would cause photochemical damage to the sample, especially living cells. Fortunately, in 1914 Stanislav Von Prowazek showed for the first time that fluorophores could be functionalized and bound to living cells [4] allowing the excitation and emission wavelengths to be engineered independently of the sample’s natural properties. Therefore, fluorescent tags quickly became standard practice in fluorescence microscopy allowing the excitation energy required to induce fluorescence to be significantly reduced. In turn, this decreased photochemical degradation of the sample by moving from ultraviolet to visible excitation wavelengths, which was extremely advantageous for biological studies allowing for sample integrity to be maintained especially for live samples. As a result, fluorescence microscopy has become one of the most widely utilized techniques in biological sciences. Figure 2 below shows an example of a cell where the three fluorophores where functionalized to attached themselves to the actin, mitochondria, and the nucleus [5].

The early generations of fluorescence microscopes utilized filtered atomic emission lamps, but most modern fluorescence microscopes employ single-mode lasers as the excitation source of choice. Solid-state lasers are far more reliable than gas discharge lamps, and their TEM00 beam profile provides the ability to be collimated and focused through complex optical geometries and focused down to a diffraction limited spot. This is especially important for confocal microscopes where the excitation source must be focused down through a pinhole to improve the z-resolution of the system. Lasers provide excellent spectral selectivity for exciting fluorophores, but unlike gas emission lamps, where the filters can be swapped to produce different excitation lines if a user wants to utilize a wide range of fluorophores in the same set-up, multiple laser sources will be required.

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CW Solid-State Laser Sources for the Life Sciences: An Introduction to Selecting a Laser Module for your Experiment or Instrument.

As lasers become more commonplace for system integrators and research labs, many times the engineer or researcher knows the key parameters needed, but has difficulties sorting out the less important parameters, which could have a significant impact on cost, the experiment, or overall performance of the system. We will present what the important considerations are when selecting the laser source and how to define the specifications to insure you get the right laser for your application. Each application, and even the setup, will require specific parameters, if you are unsure, we will define the basic specifications needed for your experiment or your instrument.

Read the full article here.

How Can We Help?

With over 25 years experience providing confocal microscopy lasers to researchers and OEM integrators working in various markets and applications, and 1000s of units fielded, we have the experience to ensure you get the right product for the application. Working with RPMC ensures you are getting trusted advice from our knowledgeable and technical staff on a wide range of laser products.  RPMC and our manufacturers are willing and able to provide custom solutions for your unique application.

If you have any questions, or if you would like some assistance please Contact Us here. Furthermore, you can email us at info@rpmclasers.com to talk to a knowledgeable Product Manager.

Alternatively, use the filters on this page to assist in narrowing down the selection of confocal microscopy lasers for sale. Finally, head to our Knowledge Center with our Lasers 101 page and Blogs, Whitepapers, and FAQ pages for further, in-depth reading.

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