A systematic comparison of sitting and hanging-drop crystallization using traditional and cross-diffusion microbatch crystallization plates

Hou, Hai; Miao, Shun; Hu, Shan-Yang; Zhang, Bin; Chen, Zhonghao; Yin, Da‐Chuan

doi:10.1016/j.jcrysgro.2019.05.011

Cited by 10 publications

(3 citation statements)

References 21 publications

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“…Considering the effect of gravity on the droplet, hanging droplet method was regarded as a common strategy for micro-droplet crystallization (especially for biomacromolecule crystal preparation) [49][50][51][52] . In this study, based on the virtues of sessile platform and CFDC, we inverted the sitting CFDC (120°) to construct a hanging CFDC on the reversed sessile platform (Fig.…”

Section: Resultsmentioning

confidence: 99%

Precise crystal regulation and harvest by constructing confined flexible droplet crystallizer on 3D-printed sessile platform

Yuan

Jiang

Niu

et al. 2021

Preprint

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Precise regulation of the mass-heat-momentum transfer carried great implications to the crystallization behavior in microscale droplet, which is one of the core concerns in crystal engineering. Herein, a confined flexible droplet crystallizer (CFDC) was facilely constructed on designed 3D-printed, matrix-type regular sessile platforms. Owing to the flexible morphology of CFDC, the synergized micro-flows generated by the integration of capillary flow and Marangoni flow in CFDC could be precisely controlled, which was validated from both experimental and simulative aspects. Standard cubic crystal was manufactured at the sessile platform center after evaporating an optimized CFDC (with 120° featured angle), where the synergized micro-flows created the perfect flow zone to balance the mass transfer and particle location. In addition, larger standard cubic crystal could be obtained via simply scale-up the platform (16 times) or supplementary feeding (secondary to forth feeding), manifesting great potential for high-quality crystal parallel preparation and harvest.

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Section: Resultsmentioning

confidence: 99%

Precise crystal regulation and harvest by constructing confined flexible droplet crystallizer on 3D-printed sessile platform

Yuan

Jiang

Niu

et al. 2021

Preprint

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“…The HD approach shares similarities with three-dimensional (3D) cell culture [31][32][33] and HD protein crystallization methods. [34][35][36][37] In both methods, the HD arrangement is employed to avoid the direct contact of sample with container surface, minimize the complicated sample/solid interactions, and thus improve the spheroid formation and crystal growth, respectively. In this study, the HD arrangement is used to keep tissue pixel away from nanowell surfaces to improve the accessibility of the extraction/digestion buffer to tissue sample, and thus to enhance the sample recovery of spatial proteomics.…”

Section: Introductionmentioning

confidence: 99%

Hanging drop sample preparation improves sensitivity of spatial proteomics

et al. 2022

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“…Specifically, we developed a hangingdrop (HD)-based sample preparation approach by simply positioning the nanoPOTS chip upside-down during protein extraction and tryptic digestion steps. The HD approach is shares similarities with three-dimensional (3D) cell culture [31][32][33] and HD protein crystallization methods [34][35][36][37] . In both methods, the HD arrangement is employed to avoid the direct contact of sample with container surface, minimize the complicated sample/solid interactions, and thus improve the spheroid formation and crystal growth, respectively.…”

Section: Introductionmentioning

confidence: 99%

Hanging-droplet sample preparation improves sensitivity of spatial proteomics

Kwon

Piehowski

Zhao

et al. 2022

Preprint

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Spatial proteomics holds great promise for revealing tissue heterogeneity in both physiological and pathological conditions. However, one significant limitation of most spatial proteomics workflows is the requirement of large sample amounts that blurs cell-type-specific or microstructure-specific information. In this study, we developed an improved sample preparation approach for spatial proteomics and integrated it with our previously-established laser capture microdissection (LCM) and microfluidics sample processing platform. Specifically, we developed a hanging drop (HD) method to improve the sample recovery by positioning a nanowell chip upside-down during protein extraction and tryptic digestion steps. Compared with the commonly-used sitting-drop method, the HD method keeps the tissue voxel away from the container surface, and thus improves the accessibility of the extraction/digestion buffer to tissue sample. The HD method can increase the MS signal by 7 fold, leading to a 66% increase in the number of identified proteins. An average of 721, 1489, and 2521 proteins can be quantitatively profiled from laser-dissected 10-µm-thick mouse liver tissue voxels with areas of 0.0025, 0.01, and 0.04 mm2, respectively. The improved system was further validated in the study of cell-type-specific proteomes of mouse uterine tissues.

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A systematic comparison of sitting and hanging-drop crystallization using traditional and cross-diffusion microbatch crystallization plates

Cited by 10 publications

References 21 publications

Precise crystal regulation and harvest by constructing confined flexible droplet crystallizer on 3D-printed sessile platform

Precise crystal regulation and harvest by constructing confined flexible droplet crystallizer on 3D-printed sessile platform

Hanging drop sample preparation improves sensitivity of spatial proteomics

Hanging-droplet sample preparation improves sensitivity of spatial proteomics

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