Protein-crystal detection with a compact multimodal multiphoton microscope

Cheng, Qing di; Chung, Hsiang Yu; Schubert, Robin; Chia, Shih Hsuan; Mudogo, Célestin Nzanzu; Kärtner, Franz X.; Chang, Guoqing; Betzel, Christian

doi:10.1038/s42003-020-01275-8

Cited by 15 publications

(5 citation statements)

References 35 publications

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“…Such development would first require further populating the parametric plot to determine the limits of multiphoton-based classification. From a practical point of view, the ongoing advances in compact multimodal multiphoton microscopes suitable for clinical use 48 and the simple sample preparation scheme shown here indicate that there is scope to optimize a multiphoton classification scheme in a high-throughput clinical setting.…”

Section: ■ Conclusionmentioning

confidence: 99%

Kidney Stone Classification Using Multimodal Multiphoton Microscopy

Gleeson,

Morizet,

Mahou

et al. 2023

ACS Photonics

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Kidney stones are a common form of nephrolithiasis, affecting up to 15% of the world's population with a high probability of recurrence. These stones exhibit various chemical compositions and crystalline forms associated with different etiologies. Classification of the stones' components is necessary to optimize treatment and suggest lifestyle changes to reduce the risk of recurrence. Current characterization methods usually require extensive sample preparation or are too detailed for the needs of a high-throughput laboratory. In this article, we present a kidney stone component classification scheme based on the multiphoton response of crushed samples that is label-free, requires minimal sample amounts, and simple preparation. We measure two-photon excited fluorescence, which is sensitive to protein content, second-harmonic generation, which is sensitive to crystalline symmetry, and polarization-resolved third-harmonic generation (pTHG), which is sensitive to crystal heterogeneity and birefringence. The combination of these three contrast modes can distinguish different materials, specifically calcium oxalate in monohydrate (COM), dihydrate (COD), or amorphous forms, cystine, and carbonate apatite. In addition, pTHG images have the potential to distinguish between COM and COD fragments and to provide information on the submicron organization of carbonate apatite fragments.

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Section: ■ Conclusionmentioning

confidence: 99%

Kidney Stone Classification Using Multimodal Multiphoton Microscopy

Gleeson,

Morizet,

Mahou

et al. 2023

ACS Photonics

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“…For example, second-order nonlinear optical imaging [38] or its complement two-photon-excited UV fluorescence microscopy [39] has many advantages when it comes to robustness in the presence of noise, e.g., being able to distinguish between protein crystals and salt crystals. However, even recent iterations require elaborate experimental setups and data acquisition times of multiple seconds per image [40], rendering them impractical for use in stirred industrial bioreactors. Additionally, image analysis and the extraction of crystal size distributions are still left as challenges, especially in systems of high protein crystal density.…”

Section: Microscopic Image Analysismentioning

confidence: 99%

Recent Advances in the Monitoring of Protein Crystallization Processes in Downstream Processing

Walla,

Bischoff,

Corona Viramontes

et al. 2023

Crystals

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Protein crystallization is nowadays performed at the micro to macro scale in academia and industry, being particularly interesting for pharmaceutical applications. Protein crystallization offers an attractive alternative to chromatography as a downstream processing step in the biotechnology industry, but also in the food and chemical industries. Monitoring of the protein crystallization process is required to understand the crystal growth mechanism and to obtain the information necessary for efficient process control, which needs to comply with the critical quality attributes of the product. Since a wide range of monitoring techniques have already been developed and established, this review focuses on the recent advances of selected techniques in monitoring protein crystallization processes such as the focused beam reflectance method (FBRM), and machine learning-based image analysis for solid-phase monitoring, as well as the spectroscopic methods for liquid-phase monitoring, such as attenuated total reflectance Fourier transform infrared (ATR-FTIR) and UV/Vis spectroscopy.

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“…Many related technologies have been developed for better SX practices, including crystal detection technology (Cheng et al, 2020), in situ diffraction devices and sample delivery systems. The sample delivery system is one of the key links in the implementation of SX research, and it is also an important tool for diversified experiments using SX technology.…”

Section: Introductionmentioning

confidence: 99%

Microfluidic rotating-target device capable of three-degrees-of-freedom motion for efficient in situ serial synchrotron crystallography

Zhao¹,

Wang²,

Xiao³

et al. 2023

J Synchrotron Rad

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There is an increasing demand for simple and efficient sample delivery technology to match the rapid development of serial crystallography and its wide application in analyzing the structural dynamics of biological macromolecules. Here, a microfluidic rotating-target device is presented, capable of three-degrees-of-freedom motion, including two rotational degrees of freedom and one translational degree of freedom, for sample delivery. Lysozyme crystals were used as a test model with this device to collect serial synchrotron crystallography data and the device was found to be convenient and useful. This device enables in situ diffraction from crystals in a microfluidic channel without the need for crystal harvesting. The circular motion ensures that the delivery speed can be adjusted over a wide range, showing its good compatibility with different light sources. Moreover, the three-degrees-of-freedom motion guarantees the full utilization of crystals. Hence, sample consumption is greatly reduced, and only 0.1 mg of protein is consumed in collecting a complete dataset.

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Protein-crystal detection with a compact multimodal multiphoton microscope

Cited by 15 publications

References 35 publications

Kidney Stone Classification Using Multimodal Multiphoton Microscopy

Kidney Stone Classification Using Multimodal Multiphoton Microscopy

Recent Advances in the Monitoring of Protein Crystallization Processes in Downstream Processing

Microfluidic rotating-target device capable of three-degrees-of-freedom motion for efficient in situ serial synchrotron crystallography

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