Subcellular Imaging of Liquid Silicone Coated-Intestinal Epithelial Cells

Abstract

Surface contamination and the formation of water bridge at the nanoscopic contact between an atomic force microscope tip and cell surface limits the maximum achievable spatial resolution on cells under ambient conditions. Structural information from fixed intestinal epithelial cell membrane is enhanced by fabricating a silicone liquid membrane that prevents ambient contaminants and accumulation of water at the interface between the cell membrane and the tip of an atomic force microscope. The clean and stable experimental platform permits the visualisation of the structure and orientation of microvilli present at the apical cell membrane under standard laboratory conditions together with registering topographical features within a microvillus. The method developed here can be implemented for preserving and imaging contaminant-free morphology of fixed cells which is central for both fundamental studies in cell biology and in the emerging field of digital pathology.

Label-Free Digital Holotomography Reveals Ibuprofen-Induced Morphological Changes to Red Blood Cells

“…For DHTM imaging of ibuprofen-treated blood smears, 25 μL of silicone oil (5 cSt, Merk Millipore) was added to the smear and a coverslip was placed on top and sealed with nail varnish (SI Appendix, Figure S11). Silicone oil was previously demonstrated to be a protective layer of cellular structures to conduct high-resolution imaging under standard laboratory conditions by circumventing the buildup of hydrocarbon and ambient contaminants …”

Spatial organization of protein aggregates on red blood cells as physical biomarkers of Alzheimer’s disease pathology

“…After initial surface analysis, the selected samples (blood smears samples that required more detailed investigation) were coated with silicone oil to prevent ambient contamination. For AFM measurements on silicone liquid-coated blood smear samples, identical materials (silicone oil purchased from Sigma-Aldrich, 317677), deposition technique (spray deposition using Harder and Steenbeck dual-action evolution spray gun), and imaging parameters as previously reported by Nirmalraj et al ( 68 ) for resolving fixed and dried epithelial cells were adopted here. For image processing, the raw AFM data were analyzed using Nanoscope analysis 1.9 (Bruker) and subjected to first-order flattening before extraction of height profile and surface roughness values from the topographic information.…”

The frontier of live tissue imaging across space and time

“…In the peritoneal cavity, different organs, including the small intestine, colon, liver, kidney, spleen, and pancreas, have been visualized at single-cell resolution using various abdominal windows (Bernier-Latmani and Petrova, 2016). Cell vitality and apoptosis, fluid transport, receptor-mediated endocytosis, blood flow, and recruitment of immune cells have been characterized using these intravital approaches (Kitamura et al, 2017;Nirmalraj et al, 2018;Sedin et al, 2019;Small et al, 2016;Zhang et al, 2014). The dynamics of the intestinal stem cell niche have been visualized using Lgr5-confetti mice with Lgr5+ stem cell labeling (Ritsma et al, 2014).…”