Ms. Anna Essaulova Profile

Anna Essaulova

at Modulight Corp

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Publications (2)

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Proceedings Article | 13 March 2024 Poster + Paper

Hyperspectral imaging for flow cytometry

A. Essaulova, R. Perttilä, A. Colucci, P. Uusimaa

Proceedings Volume 12834, 1283409 (2024) https://doi.org/10.1117/12.3003862

KEYWORDS: Cameras, Hyperspectral imaging, Flow cytometry, Particles, Imaging systems, Biological imaging, Light sources and illumination, Fluorescence, Biological samples, RGB color model

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Flow cytometry is a widely used analysis technique in biomedical sciences. It has found extensive utilization in both clinical diagnostics and cutting-edge biological research. As the method has been gaining greater recognition, its underlying technologies have undergone rapid development to further expand its range of applications. A notable trend is the introduction of imaging modalities to flow cytometry to expand the information content of the analyzed sample.

The introduction of a camera component to the already well-established detectors, such as photo multiplier tubes (PMTs) or avalanche photo diodes (APDs), adds intricacy to the arrangement of optical subsystem in flow cytometer. Moreover, it brings forth additional requirements for effectively coordinating information capture among different detector types. An appealing alternative to address this challenge is hyperspectral imaging – a technique which enables capturing of the spatial and spectral information simultaneously. Yet, there has not been much research performed to study applications of hyperspectral imaging in combination with narrow bandwidth illumination commonly used in flow cytometry.

In this work, we investigate the applicability of hyperspectral imaging to flow cytometrical systems, where a multiple wavelength laser system is utilized for sample illumination. A four-wavelength laser illumination platform developed by Modulight Corporation is utilized as the light source. Our main objective is to assess the hyperspectral imaging component's ability to distinguish between the illuminating light and the fluorescence emitted by the sample. Furthermore, we carefully evaluate the quality of the obtained hyperspectral images and explore the potential to differentiate samples based on the collected spatial data.

Proceedings Article | 12 March 2024 Presentation + Paper

High-power VECSEL with intracavity UV generation for flow cytometry applications

K. Nechay, R. Perttilä, A. Essaulova, P. Uusimaa

Proceedings Volume 12853, 1285306 (2024) https://doi.org/10.1117/12.3002263

KEYWORDS: Ultraviolet radiation, Vertical external cavity surface emitting lasers, Flow cytometry, Laser resonators, Sum frequency generation, Semiconductor lasers, Frequency conversion, Second harmonic generation, Crystals, Nonlinear crystals

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Flow cytometry is a fundamental and powerful technique in the biomedical sciences which heavily depends on excitation lasers for imaging fluorescent markers to perform cell sorting and analysis. Modern multicolor flow cytometers incorporate several excitation lasers, typically 405 nm, 488 nm, 561 nm, and 640 nm allowing simultaneous cell characterization. Ultraviolet (UV) lasers, namely 355 nm, became indispensable flow cytometry excitation sources with the development of the Brilliant Ultraviolet (BUV) fluorochromes enabling high-dimensional analysis. Primary UV source for BUV excitation is represented by frequency tripled Nd:YVO₄ diode-pumped solid-state lasers (DPSSL) which constitute the single most expensive component of high-end flow cytometers. We demonstrate high-power UV generation from cascade intracavity frequency conversion based on the optically pumped vertically-external-cavity surface-emitting lasers (VECSELs). The active semiconductor gain medium is designed for operation around 1060 nm and is comprised of InGaAs quantum wells. The frequency conversion is implemented in a compact V-shaped cavity with the non-linear lithium triborate (LBO) crystals placed at the waist of the cavity mode in order to enhance conversion efficiency. This relatively novel and cost-effective semiconductor laser platform offers high-power, tunable, low-noise emission with excellent beam quality, important for flow cytometry applications. VECSEL platform can fill the spectral gaps where semiconductor diode laser technology is not available, namely green, and green-yellow spectral regions, and at the same time serving as a cost-effective replacement of DPSSL in the UV and DUV spectral bands.

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