Biological Engineering

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    Protocols in Current Issue
    Preparation of Cas9 Ribonucleoproteins for Genome Editing
    Authors:  Sheng-Wei Lin, Viet Quoc Nguyen and Steven Lin, date: 05/20/2022, view: 829, Q&A: 0
    [Abstract]

    Genome editing by the delivery of pre-assembled Cas9 ribonucleoproteins (Cas9 RNP) is an increasingly popular approach for cell types that are difficult to manipulate genetically by the conventional plasmid and viral methods. Cas9 RNP editing is robust, precise, capable of multiplexing, and free of genetic materials. Its transient presence in

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    Immunomagnetic Isolation and Enrichment of Microvascular Endothelial Cells from Human Adipose Tissue
    [Abstract]

    Human adipose tissue-resident microvascular endothelial cells are not only garnering attention for their emergent role in the pathogenesis of obesity-related metabolic disorders, but are also of considerable interest for vascular tissue engineering due, in part, to the abundant, accessible, and uniquely dispensable nature of the tissue. Here, we

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    Activation of Mitochondrial Ca2+ Oscillation and Mitophagy Induction by Femtosecond Laser Photostimulation
    Authors:  Xiaoying Tian and Hao He, date: 04/05/2022, view: 328, Q&A: 0
    [Abstract]

    Ultra-precise stimulation solely to individual mitochondria, without any influence to the whole cell, is quite difficult by traditional biochemical reagents. In mitophagy research, the mitochondria and even the whole cell usually suffer irreversible and great damage caused by treatment with potent chemicals. In this protocol, we present the

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    Preparation, Characterization, and Cell Uptake of PLGA/PLA-PEG-FA Nanoparticles
    Authors:  Heng Yang, Junsik Sung, Zahra Alghoul and Didier Merlin, date: 04/05/2022, view: 362, Q&A: 0
    [Abstract]

    Oral administration of colon-targeting nanoformulations holds many advantages over the systemic delivery of free drugs, or traditional nontargeting formulations in the treatment of ulcerative colitis (UC). Currently, the most conventional method for constructing colon-targeting drug delivery systems (DDS) is by integrating the biocompatible

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    Quantitative and Anatomical Imaging of Human Skin by Noninvasive Photoacoustic Dermoscopy
    Authors:  Zhiyang Wang, Fei Yang, Wuyu Zhang and Sihua Yang, date: 04/05/2022, view: 378, Q&A: 0
    [Abstract]

    Imaging plays a vital role in the diagnosis and treatment of skin diseases. However, pure optical imaging technique is limited to the visualization of superficial skin tissues. Ultrasonic imaging technique can detect deep tissues, but it lacks detailed information on microscopic pathological structures. Photoacoustic imaging is an advanced

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    Ex-vivo Skin Permeability Tests of Nanoparticles for Microscopy Imaging
    Authors:  Nethanel Friedman, Sharon Merims, Jhonatan Elia and Ofra Benny, date: 04/05/2022, view: 386, Q&A: 0
    [Abstract]

    Delivery of drugs through the skin is a major challenge in the field of drug delivery systems. Quantification of materials, and specifically nanoparticles, within the skin layers is essential for the development of advanced topical and transdermal delivery systems. We have developed a technique for ex-vivo segmentation and evaluation of human

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    A Rapid Protocol for Direct Isolation of Osteoclast Lineage Cells from Mouse Bone Marrow
    [Abstract]

    Osteoclast lineage cells (OLCs), including osteoclast precursors (OCPs) and mature osteoclasts (MOCs), participate in bone remodeling and mediate pathologic bone loss. Thus, it is essential to obtain OLCs for exploring their molecular features in both physiological and pathological conditions in vivo. However, the conventional protocols for

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    An in vitro Blood-brain Barrier Model to Study the Penetration of Nanoparticles
    Authors:  Sezin Aday, Wen Li, Jeffrey M. Karp and Nitin Joshi, date: 02/20/2022, view: 2560, Q&A: 0
    [Abstract]

    The blood-brain barrier (BBB), a crucial protection mechanism in the central nervous system (CNS), is a selective barrier comprised of endothelial cells. It hampers the development of therapeutic and diagnostic tools for neurological diseases due to the poor penetration of most of these agents. Rationally engineered nanoparticles (NP) can

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    An Alternative Technique for Monitoring the Live Interaction of Monocytes and Tumor Cells with Nanoparticles in the Mouse Lung
    Authors:  Fernanda Ramos-Gomes, Nathalia Ferreira, Frauke Alves and M. Andrea Markus, date: 01/20/2022, view: 1187, Q&A: 0
    [Abstract]

    Nanomaterials are increasingly used for the diagnosis and treatment of cancer, including lung cancer. For the clinical translation of nano-based theranostics, it is vital to detect and monitor their accumulation in the tumor, as well as their interaction with tumor, immune cells, and the tumor microenvironment (TME). While high resolution

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    A Simple Technique for Direct Immobilization of Target Enzymes from Cell Lysates Based on the SpyTag/SpyCatcher Spontaneous Reaction
    Authors:  Li-xi Cai, Yuan-qing Lin, Yun-meng Chu, Xiao-pin Chen, Li-xing Liu, Ming Zhang and Guang-ya Zhang, date: 01/05/2022, view: 1463, Q&A: 0
    [Abstract]

    Many of the current methods for enzyme purification and immobilization suffer from several drawbacks, such as requiring tedious multistep procedures or long preparation, and being environmentally unfriendly, due to the chemicals and conditions involved. Thus, a simple technique for direct purification and immobilization of target enzymes from cell

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