2020
DOI: 10.3390/ijms21176373
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Developing Electron Microscopy Tools for Profiling Plasma Lipoproteins Using Methyl Cellulose Embedment, Machine Learning and Immunodetection of Apolipoprotein B and Apolipoprotein(a)

Abstract: Plasma lipoproteins are important carriers of cholesterol and have been linked strongly to cardiovascular disease (CVD). Our study aimed to achieve fine-grained measurements of lipoprotein subpopulations such as low-density lipoprotein (LDL), lipoprotein(a) (Lp(a), or remnant lipoproteins (RLP) using electron microscopy combined with machine learning tools from microliter samples of human plasma. In the reported method, lipoproteins were absorbed onto electron microscopy (EM) support films from diluted plasma … Show more

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Cited by 3 publications
(3 citation statements)
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References 68 publications
(97 reference statements)
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“…To prevent artifacts from drying or vitrication of liposomes and VLPs for TEM, a methylcellulose matrix-assisted drying process was used as a structural support for the particles before negative staining with uranyl acetate. 80,81 Briey, 3 mm copper grids with carbon lm (CF200-CU-50 200 mesh) were glow discharged (PELCO easiGlow), then grids were inverted on drops of sample and incubated at room temperature for 60 minutes. The grids were washed with 1× PBS for 1 minute, xed for 5 minutes with 4% paraformaldehyde + 2.5% glutaraldehyde in 0.1 M sodium cacodylate, and then stained with 2% uranyl acetate for 5 minutes.…”
Section: Transmission Electron Microscopymentioning
confidence: 99%
See 1 more Smart Citation
“…To prevent artifacts from drying or vitrication of liposomes and VLPs for TEM, a methylcellulose matrix-assisted drying process was used as a structural support for the particles before negative staining with uranyl acetate. 80,81 Briey, 3 mm copper grids with carbon lm (CF200-CU-50 200 mesh) were glow discharged (PELCO easiGlow), then grids were inverted on drops of sample and incubated at room temperature for 60 minutes. The grids were washed with 1× PBS for 1 minute, xed for 5 minutes with 4% paraformaldehyde + 2.5% glutaraldehyde in 0.1 M sodium cacodylate, and then stained with 2% uranyl acetate for 5 minutes.…”
Section: Transmission Electron Microscopymentioning
confidence: 99%
“…80,81 Briey, 3 mm copper grids with carbon lm (CF200-CU-50 200 mesh) were glow discharged (PELCO easiGlow), then grids were inverted on drops of sample and incubated at room temperature for 60 minutes. The grids were washed with 1× PBS for 1 minute, xed for 5 minutes with 4% paraformaldehyde + 2.5% glutaraldehyde in 0.1 M sodium cacodylate, and then stained with 2% uranyl acetate for 5 minutes.…”
mentioning
confidence: 99%
“…EV were imaged in a JEOL 1200EX electron microscope equipped with a GATAN Orius 200 digital camera. EV were selected for quantification using unbiased counting rule applied to a scanning band [36] and calliper diameters measured either directly on screen or estimated using stereological techniques as previously described [33,36].…”
Section: Immuno-electron Microscopymentioning
confidence: 99%