Raman Cryomicroscopic Imaging and Sample Holder for Spectroscopic Subzero Temperature Measurements

Yu, Guanglin; Li, Rui; Hubel, Allison

doi:10.1007/978-1-0716-0783-1_14

Cited by 4 publications

(2 citation statements)

References 21 publications

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“…Cell samples were prepared in the form of single-cell suspension and frozen at a controlled rate for analysis by Raman spectroscopy following a previously published method ( Li et al, 2020 ; Yu et al, 2021 ). Specifically, defined cooling rates of −1, −3, and −5°C/min and ice nucleation temperatures of −4°C and −8°C were used as variables of the freezing profile.…”

Section: Methodsmentioning

confidence: 99%

Differentiation of Human iPS Cells Into Sensory Neurons Exhibits Developmental Stage-Specific Cryopreservation Challenges

Walsh

Truong

et al. 2021

Front. Cell Dev. Biol.

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Differentiation of human induced pluripotent stem cells (hiPSCs) generates cell phenotypes valuable for cell therapy and personalized medicine. Successful translation of these hiPSC-derived therapeutic products will rely upon effective cryopreservation at multiple stages of the manufacturing cycle. From the perspective of cryobiology, we attempted to understand how the challenge of cryopreservation evolves between cell phenotypes along an hiPSC-to-sensory neuron differentiation trajectory. Cells were cultivated at three different stages to represent intermediate, differentiated, and matured cell products. All cell stages remained ≥90% viable in a dimethyl sulfoxide (DMSO)-free formulation but suffered ≥50% loss in DMSO before freezing. Raman spectroscopy revealed higher sensitivity to undercooling in hiPSC-derived neuronal cells with lower membrane fluidity and higher sensitivity to suboptimal cooling rates in stem cell developmental stages with larger cell bodies. Highly viable and functional sensory neurons were obtained following DMSO-free cryopreservation. Our study also demonstrated that dissociating adherent cultures plays an important role in the ability of cells to survive and function after cryopreservation.

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Section: Methodsmentioning

confidence: 99%

Differentiation of Human iPS Cells Into Sensory Neurons Exhibits Developmental Stage-Specific Cryopreservation Challenges

Walsh

Truong

et al. 2021

Front. Cell Dev. Biol.

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“…Raman spectroscopy has emerged as a powerful tool to investigate the freezing responses of various types of cells (Yu et al, 2021). This method is label-free and has high chemical specificity and spatial resolution.…”

Section: Introductionmentioning

confidence: 99%

Characterizing cellular membrane partitioning of DMSO using low-temperature Raman spectroscopy

Louwagie

Wagner²,

Li³

et al. 2023

Front. Mol. Biosci.

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Additives that help cells survive the stresses of freezing and thawing are known as cryoprotective agents (CPAs). Two different types of CPAs have been identified: penetrating and non-penetrating. Common penetrating CPAs include dimethylsulfoxide (DMSO) and glycerol. The location of a CPA (intracelluar or extracellular) is important for understanding the molecular mechanisms of action for the agent. Low-temperature Raman spectroscopy is a label-free method of detecting the location of CPAs at low temperature with high spatial resolution and chemical specificity. To this end, cells cryopreserved in DMSO using a variety of cooling rates and DMSO concentrations and imaged using Raman spectroscopy were analyzed using automated image analysis to determine the partitioning ratio (concentration of DMSO outside/concentration of DMSO inside the cell). The partitioning ratio was roughly 1 for Jurkat cells frozen at 1°C/min in varying concentrations of DMSO with the exception of 1% DMSO which had a partitioning ratio of 0.2. The partitioning ratio increased from 1 to 1.3 as the cooling rate increased from 1°C to 5°C/min. Different cell types, specifically sensory neurons cells and human induced pluripotent stem cells, exhibited differences in partitioning ratio when frozen in 10% DMSO and 1°C/min suggesting that differences in freezing response may result from differences in solute partitioning. The presence of intracellular ice changed the distribution of DMSO inside the cell and also the partitioning ratio.

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Cryoconveyor Protocols in Correlative Light and Electron Microscopy: From Multilevel Imaging to Modeling the Biophysical Effects and “Cryotheranostics”

Gradov

2023

Moscow Univ. Biol.Sci. Bull.

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Raman Cryomicroscopic Imaging and Sample Holder for Spectroscopic Subzero Temperature Measurements

Cited by 4 publications

References 21 publications

Differentiation of Human iPS Cells Into Sensory Neurons Exhibits Developmental Stage-Specific Cryopreservation Challenges

Differentiation of Human iPS Cells Into Sensory Neurons Exhibits Developmental Stage-Specific Cryopreservation Challenges

Characterizing cellular membrane partitioning of DMSO using low-temperature Raman spectroscopy

Cryoconveyor Protocols in Correlative Light and Electron Microscopy: From Multilevel Imaging to Modeling the Biophysical Effects and “Cryotheranostics”

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