⭐ High Impact

Visualizing cell architecture and molecular location using soft x-ray tomography and correlated cryo-light microscopy.

McDermott Gerry, Le Gros Mark A, Larabell Carolyn A

📰 Annual review of physical chemistry 📅 2012 📊 76 citations

Abstract

Living cells are structured to create a range of microenvironments that support specific chemical reactions and processes. Understanding how cells function therefore requires detailed knowledge of both the subcellular architecture and the location of specific molecules within this framework. Here we review the development of two correlated cellular imaging techniques that fulfill this need. Cells are first imaged using cryogenic fluorescence microscopy to determine the location of molecules of interest that have been labeled with fluorescent tags. The same specimen is then imaged using soft X-ray tomography to generate a high-contrast, 3D reconstruction of the cells. Data from the two modalities are then combined to produce a composite, information-rich view of the cell. This correlated imaging approach can be applied across the spectrum of problems encountered in cell biology, from basic research to biotechnological and biomedical applications such as the optimization of biofuels and the development of new pharmaceuticals.

🔬 Techniques

🧬 Organisms

✨ Fluorophores

GFP

🧪 Sample Preparation

💻 Software Details

Image Analysis:
inForm

🏛️ Research Organizations (ROR)

Affiliated research institutions:

📊 Figures

Figure 1

( a ) The optical configuration of a bend magnetu2013based transmission soft X-ray microscope (XM-2) located at the Advanced Light Source, Berkeley, California. The specimen sits between the objective...

Figure 2

( a ) Soft X-ray tomographic reconstruction of the yeast Schizosaccharomyces pombe . Two orthoslices are shown from different regions in the tomographic reconstruction of a cell ( left, center ) illus...

Figure 3

Protein localization using immunolabeling methods. Plasmodium falciparum -infected erythrocytes expressing green fluorescent protein (GFP) chimeras of a Maureru2019s cleft resident protein, the membra...

Figure 4

An X-ray tomographic reconstruction of mouse adenocarcinoma cells showing many subcellular organelles. Abbreviations: ER, endoplasmic reticulum; L, lysosomes; M, mitochondria; NM, nuclear membrane; NP...

Figure 5

Correlated soft X-ray tomography and cryo-light imaging (wide-field fluorescence). ( a ) The vacuoles fluorescently labeled and imaged by cryo-light microscopy. ( b,c ) Slices through the volumetric r...

Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.

🏛️ Imaging Facility

🏛️ University of California

💬 Discussion

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