Next-Generation Laser Scanning Multiphoton Microscopes are Turnkey, Portable, and Industry-Ready


Michael E. Holmes,1 * Stefanie Kiderlen,2 *mholmes@two-levelsystems.com


Abstract: The Prospective Instruments MPX-series is a turnkey com- pact multimodal microscope enabling advanced multiphoton imaging without requiring a laboratory or optical bench. One of the key design features is the ultrafast fiber laser engine integrated into the free mov- ing scan head, which is lightweight, rugged, and allows the ultimate imaging flexibility. A modular design concept allows the user to explore a wide variety of biological applications and measurements, without compromising performance, in any indoor working space. In contrast, standard setups are non-transportable, utility-demanding, and the complexity can be complicated, therefore degrading the value and adding significant initial, short- and long-term expenses. Every clini- cal and biological researcher should have access to high-quality reli- able optical microscopy modalities without being hardware experts. In this article we outline the demand and design of the MPX-microscope and present multiphoton imaging results from experiments in various configurations. There are no existing instruments on the market that are portable and combine easily switchable modes in one composite industry-ready device for life science, clinical, and pharmacological research.


Keywords: biophotonics, multiphoton microscopy, multimodal microscopy, epi-fluorescence, label-free imaging


Introduction Multiphoton microscopy uses a highly focused laser beam


to produce nonlinear optical effects based on the interaction of multiple photons arriving simultaneously at a molecule. Terefore, the intensity of the generated signal does not increase linearly with the number of irradiated photons, but with the square (for two-photon effects) or the third power (for three- photon effects), and the signal is intrinsically confocal [1]. Using wavelengths in the near infrared (NIR) range, multiphoton imaging allows similar resolution as conventional confocal microscopy, but with much higher tissue penetration depths of up to 1 mm or beyond [2,3]. Currently available commercial laser scanning multipho-


ton microscopes (LSMM) have significant drawbacks that make systems difficult to incorporate in clinical environments, or non-accessible for many researchers. Typically, these systems include large vibration-isolation optical tables, require exces- sive utilities to operate, and are difficult to maintain. Te two main components, the scan head (including beam-steering optics) and ultrafast laser source, are oſten manufactured by separate vendors. Te upfront cost for a complete microscope can be >$500,000 and requires a heſty yearly service contract and costly maintenance overtime. Tese expenses, along with ease of use and mobility, are challenging factors in making the devices accessible. Tis drives the need for a compact and por- table imaging tool with rapid scanning and imaging acquisition time, while maintaining cellular resolution. Prospective Instru- ments (Dornbirn, Austria) has recently created a device with a compact, fast-scanning, and flexible front-end that addresses


16 doi:10.1017/S1551929522000657


these challenges (Figure 1). Te entire microscope consists of two parts that are connected through a flexible umbilical cord. Te control box is on wheels and incorporates a built-in PC together with other electronic and photonic devices. Intelligent engineering of a modular and efficient design based on fiber technology make the system cost-efficient and sustainable. Te MPX makes LSMM simple, portable, and user-friendly, even at home by non-experts. Te future for LSMM requires an instrument that can suc-


cessfully operate in different working environments outside of the laboratory. For example, in March 2020, research around the world was stalled due to the COVID-19 pandemic. How- ever, research with the MPX continued for a collaborator who was using the device in his laboratory and then transferred the entire LSMM into his home basement (Figure 1D). Stan- dard utilities like those required by a household appliance are sufficient with no special power or water requirements. Aſter relocation, the microscope was ready to make measurements immediately. Te unique usability and flexibility of the MPX does not compromise research quality and provides the same data reliability as commercially available large and very costly multiphoton systems. Tis experiment was the ultimate dem- onstration of the compactness, robustness, and portability of a complete LSMM, and probably a world first. With microscopy at home made this easy, translating LSMM imaging to the clinic is a promising next step.


Clinical Researchers Demand a Compact and Turnkey Multimodal Microscope Because of growing healthcare applications, development


of LSMMs for science and technology has been largely sup- ported by the National Institutes of Health (NIH). Detailed observation at the cellular level is critical for understanding pathogens and developing therapeutic solutions. In addition, depth-resolved noninvasive imaging of tissue in its native envi- ronment is highly beneficial. Funding for research has increased almost exponentially in the past 10 years and does not appear to be slowing. Te key medical fields benefiting include the brain and nervous system, cancer research, mental illness, and digital pathology. Improving fundamental and clinical research paves the way for prevention, recovery, and cures. As the applications grow and techniques mature, the demand escalates for easy-to- use, efficient, and multimodal microscopes. Also, it is necessary to have transportable instruments capable of delivering impact- ful insights and high-quality results. Tere are a few nonlinear optical modalities being exten-


sively considered for clinical purposes. For example, two- photon, three-photon, second-harmonic generation (SHG), third-harmonic generation (THG), coherent anti-stokes Raman


www.microscopy-today.com • 2022 May and Lukas Krainer2


1two-level systems, 501 1st Ave. North, # 901, St. Petersburg, FL 33701 2Prospective Instruments LK OG, Stadtstraße 33, 6850 Dornbirn, Austria


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