Confocal microscopy is an optical technique that allows for extremely high spatial resolution imaging by placing two conjunct pinholes in the optical path. This provides for z-axis resolution as fine as 1 micron, making it possible to measure 3D depth profiles of materials. In confocal fluorescence microscopy, a laser is focused through the first pinhole and used to excite the sample, and then the emitted light is collected and focused through the second pinhole to the detector. Most of the visible wavelengths can be used in confocal fluorescence microscopy, but it’s essential that all confocal fluorescence microscopy lasers have a high quality TEM00 beam profile. On this page, you will find a list of all of the full range of lasers used for this application, including our multi-laser combiners.
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.
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.
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.
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.
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@rpmcdev.maxdroplet4.maxburst.devto 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.
Finally, check out our Limited Supply – In Stock – Buy Now page: This page contains an ever-changing assortment of various types of new lasers at marked-down/discount prices.
The Matchbox series offers excellent performance and reliability in an ultra-compact “all-in-one” integrated laser head. They come standard with an integrated internal voltage up-conversion that allows using a 5V power supply while maintaining low noise operation. The monolithic design of the Matchbox Series laser includes thermally stabilized optics in a hermetically sealed housing, ensuring reliable and maintenance-free operation. All the Matchbox series modules include a 12-month warranty and are RoHS compliant. All of these features make them the ideal laser source for integration into commercial flow cytometers.
The LGK Series of HeNe Lasers feature a robust mechanical design, excellent beam quality, and a variety of configurations. These HeNe lasers are available in the Green, Yellow, and Red spectral ranges, and boast an excellent TEM00 beam, robust mechanical design, and a long service life of up to 30,000 hours. 543 nm, 594 nm, and 632.8 nm (633 nm) wavelengths are available, with output power levels from 0.5 mW to 20 mW.
The HL Series of laser diodes are available in a wide range of wavelengths from violet to red and infrared in support of a broad range of applications. These applications include display, medical, biosciences, industrial tools (sensor, leveler), machine vision, scanners, printers and a myriad of other applications being developed in the industry.
The LaserBoxx low noise Series is a CW diode laser module in a variety of diode wavelengths from 375 to 785nm, with output powers up to 350mW. This compact, self-contained laser module is available in turn-key or OEM versions and utilizes a proprietary alignment-free, monolithic resonator, and comes standard with a graphic user interface with remote diagnostics via USB, RS232, or direct I/O interface.
The LXCc series is the most compact and flexible all-in-one multicolor laser combiner, with up to 7 laser lines, and up to 500mW output power per line. The turnkey or OEM versions allow a large choice of lasers from 375nm up to 1064nm. The extension module provides the ultimate level of flexibility with up to 4 optical fiber outputs, integrated fast switching output ports for FRAP, or adjustable split power for light-sheet microscopy.
The R series of wavelength stabilized single mode and multimode laser diodes offer narrow wavelength spectrum in wavelengths from 633nm thru 1064nm. Package options range from components as basic as a TO-56 or 14-pin BF packaged diodes, to OEM modules including electronics, to UL/CE and IEC certified turn-key systems.