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Subcellular Fractionation of Hela Cells for Lysosome Enrichment Using a Continuous Percoll-density Gradient

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Conjugation of Fab’ Fragments with Fluorescent Dyes for Single-molecule Tracking on Live Cells

Authors: I-Ting Teng, Xiangning Bu and Inhee Chung, date: 09/20/2019, view: 50, Q&A: 0

Our understanding of the regulation and functions of cell-surface proteins has progressed rapidly with the advent of advanced optical imaging techniques. In particular, single-molecule tracking (SMT) using bright fluorophores conjugated to ...

SIRF: A Single-cell Assay for in situ Protein Interaction with Nascent DNA Replication Forks

Authors: Sunetra Roy and Katharina Schlacher, date: 09/20/2019, view: 51, Q&A: 0

The duplication of DNA is a fundamental process that is required for the transfer of the genetic information from parent to daughter cells. Aberrant DNA replication processes are associated with diverse disease phenotypes, including developmental ...

A Cell Culture Model that Mimics Physiological Tissue Oxygenation Using Oxygen-permeable Membranes

Authors: Chloe-Anne Martinez, Peter A. Cistulli and Kristina M. Cook, date: 09/20/2019, view: 78, Q&A: 0

Dissolved oxygen and its availability to cells in culture is an overlooked variable which can have significant consequences on experimental research outcomes, including reproducibility. Oxygen sensing pathways play key roles in cell growth and ...

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Conjugation of Fab’ Fragments with Fluorescent Dyes for Single-molecule Tracking on Live Cells

Authors: I-Ting Teng, Xiangning Bu and Inhee Chung, date: 09/20/2019, view: 50, Q&A: 0

[Abstract] Our understanding of the regulation and functions of cell-surface proteins has progressed rapidly with the advent of advanced optical imaging techniques. In particular, single-molecule tracking (SMT) using bright fluorophores conjugated to antibodies and wide-field microscopy methods such as total internal reflection fluorescence microscopy have ...

SIRF: A Single-cell Assay for in situ Protein Interaction with Nascent DNA Replication Forks

Authors: Sunetra Roy and Katharina Schlacher, date: 09/20/2019, view: 51, Q&A: 0

[Abstract] The duplication of DNA is a fundamental process that is required for the transfer of the genetic information from parent to daughter cells. Aberrant DNA replication processes are associated with diverse disease phenotypes, including developmental defects, ageing disorders, blood disorders such as Fanconi Anemia, increased inflammation and cancer. ...

A Cell Culture Model that Mimics Physiological Tissue Oxygenation Using Oxygen-permeable Membranes

Authors: Chloe-Anne Martinez, Peter A. Cistulli and Kristina M. Cook, date: 09/20/2019, view: 78, Q&A: 0

[Abstract] Dissolved oxygen and its availability to cells in culture is an overlooked variable which can have significant consequences on experimental research outcomes, including reproducibility. Oxygen sensing pathways play key roles in cell growth and behavior and pericellular oxygen levels should be controlled when establishing in vitro models. ...

A Robust, One-step FRET Assay for Human Heparanase

Authors: Jyothi C. Sistla and Umesh R. Desai, date: 09/05/2019, view: 208, Q&A: 0

[Abstract] Heparanase, an endo-β-D-glucuronidase, cleaves cell surface and extracellular matrix heparan sulfate (HS) chains at distinct sites and plays important biological roles including modulation of cell growth and metastasis. Although a number of different types of heparanase assays have been reported to date, most are labor intensive, complex ...

Three-dimensional Reconstruction and Quantification of Proteins and mRNAs at the Single-cell Level in Cultured Cells

Authors: Xueer Jiang, Ingrid Brust-Mascher and Li-En Jao, date: 08/20/2019, view: 536, Q&A: 0

[Abstract] Gene expression is often regulated by the abundance, localization, and translation of mRNAs in both space and time. Being able to visualize mRNAs and protein products in single cells is critical to understand this regulatory process. The development of single-molecule RNA fluorescence in situ hybridization (smFISH) allows the detection of ...

Photodynamic Therapy in a 3D Model of Ovarian Cancer

Authors: Shubhankar Nath and Kaitlin Moore, date: 08/05/2019, view: 365, Q&A: 0

[Abstract] Photodynamic therapy (PDT), is a clinically-approved light-based anti-cancer treatment modality in which a photoactivatable photosensitizer is irradiated with an appropriate wavelength of light to generate cytotoxic molecules to kill cancer cells. In this article, we describe an in vitro PDT protocol using a 3-dimensional (3D) model of ...

Evaluation of Genotoxicity by Micronucleus Assay in vitro and by Allium cepa Test in vivo

Authors: Christina N. Banti and Sotiris K. Hadjikakou, date: 07/20/2019, view: 782, Q&A: 0

[Abstract] The in vitro and in vivo genotoxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) is evaluated by the micronucleus test and the Allium cepa assay, respectively. Fetal lung fibroblast cells (MRC-5), normal human corneal epithelial cells (HCEC) and ...

Total RNA Isolation from Separately Established Monolayer and Hydrogel Cultures of Human Glioblastoma Cell Line

Authors: Manasi P Jogalekar and Elba E Serrano, date: 07/20/2019, view: 793, Q&A: 0

[Abstract] Astrocytoma is an invasive carcinoma occurring in the nervous system and currently lacks effective treatment options. A deeper understanding of the mechanisms of tumorigenesis and tumor progression is needed in order to develop novel therapeutic strategies. Recent advances in in vitro culture systems have demonstrated that the use of ...

The Chick Chorioallantoic Membrane (CAM) Assay as a Three-dimensional Model to Study Autophagy in Cancer Cells

Authors: Félice A. Janser, Patricia Ney, Marta Teixeira Pinto, Rupert Langer and Mario P. Tschan, date: 07/05/2019, view: 719, Q&A: 0

[Abstract] The chick chorioallantoic membrane (CAM) is an extra-embryonic organ and thus well accessible for seeding and harvesting 3D cell cultures. Samples from CAM assays are suitable for protein and gene expression analysis as well as for immuno-histochemical studies. Here we present the CAM assay as a possible model to study autophagy in different types ...

Capillary Nano-immunoassay for Quantification of Proteins from CD138-purified Myeloma Cells

Authors: Irena Misiewicz-Krzeminska, Isabel Isidro and Norma C. Gutiérrez, date: 06/20/2019, view: 1340, Q&A: 0

[Abstract] Protein analysis in bone marrow samples from patients with multiple myeloma (MM) has been limited by the low concentration of proteins obtained after CD138⁺ cell selection. A novel approach based on capillary nano-immunoassay could make it possible to quantify dozens of proteins from each CD138⁺ purified MM sample in an ...

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