SingleVirusTrackinginLiveCells
Single-Virus Tracking in Live CellsMichael J. Rust, Melike Lakadamyali, Boerries Brandenburg and Xiaowei Zhuang INTRODUCTIONReal-time, live-cell imaging techniques and single-particle tracking algorithms can be used to follow individual virus particles as they infect cells. This article describes the labeling of viruses and cellular structures with fluorescent probes to allow visualization in live cells. It also......阅读全文
Detection－of－BrdU－Incorporation－in－DNA－Synthesizing－Cells
Detection of BrdU Incorporation in DNA Synthesizing Cells NOTE: Bromodeoxyuridine is a known carcinogen. Propidium iodine (PI) is known to be toxic an
TOP10－chemically－competent－cells
OverviewThis protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the B
Isolation－and－Culture－of－Human－Liver－Stem－Cells
Materials and Methods1. Human hepatocytes were isolated from fresh surgical specimens of patients undergoing hepatectomies. Healthy liver tissue (
Culture－of－human－venous－smooth－muscle－cells
Culture of human venous smooth muscle cells 1. Explants of the 2 groups of skins (control subjects and patients with varicose veins) were prepared
PriCells:－Isolation－of－endothelial－progenitor－cells－(EPCs)
PriCells: Isolation of endothelial progenitor cells (EPCs) 1. Twenty-four mL venous blood was collected at each time point into Vacutainer CPT Mono
Transient－transfection－into－293T－cells
PurposeTransient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) pro
Preparation－Of－Peripheral－Blood－Cells－For－Chromosome－Analysis
实验概要Lymphocytes are differentiated cells which normally do not undergo subsequent cell divisions. By culturing lymphocytes in the presence of a mito
Transient－Transfection－of－Cos1－Cells
Transient Transfection of Cos-1 CellsNote: This protocol has been optimized for Cos-1 cells. Successful transfection of each cell type requires optimi
Isolation－and－cultivation－of－endothelial－progenitor－cells－(EPCs)
Circulating bone marrow (BM)–derived endothelial progenitor cells (EPCs) are recruited to the site of tissue regeneration and substantially contri
Metabolic－Labeling－of－Cells－with－35S
1) Transfer to a 24 wells plate the desired colonies.2) Once the cells are attached (at least 8 hours after tripsinizing them) add ~1 ml ofDME (met-,
Culture－of－Human－Embryonic－Stem－Cells－(hESC)
All cell lines are initially grown according to the supplier's protocols but we are adapting them to one simple protocol outlined below:6-well pla
Cell－cycle－analysis－of－Escherichia－coli－cells
Cell cycle analysis of Escherichia coli cellsC period = the time for a round of chromosome replicationD period = the time between the end of a round o
Differentiating－Glial－Precursor－Cells－into－Astrocytes－and－Oligodendrocytes
实验概要Glial precursor cells (GPCs), also known as glial restricted progenitors (GRP) or oligodendrocyte progenitor cells (OPCs), are cells that have
ex－vivo－expanded－endothelial－progenitor－cells
Cell Culture. 1. Total hPBMCs were isolated from blood of human volunteers by density gradient centrifugation. 2. Cells were plated on culture dishes
Cellbased－ELISA－for－primary－cells
1. The 96-well micro-plates are pre-coated with extra cellular matrix.2. Primary cells are grown in the 96-well plates. Seed 100μl of 10,000-20,000
Cryopreservation－and－Recovery－of－Mature－Differentiated－Neural－Cells
实验概要Primary neuronal cultures are indispensable in the field of neurobiology and pharmacology. Many researchers favor freshly isolated neuronal cell
Differentiate－ES－cells－into－cystic－embryoid－bodies
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.______________Day 1: Trypsinized th
HThymidine－Uptake－by－Cultured－Cells
H-Thymidine Uptake by Cultured CellsLEVEL IIMaterialsFibroblast cells in log phase growthCa, Mg free-phosphate buffered saline (PBSA)5% (w/v) Glutaral
Isolation－of－peripheral－blood－mononuclear－cells－(PBMC)
1. Acid-citrate dextrose-treated blood (450 ml) was obtained from donors. 2. PBMC were isolated by centrifugation over lymphocyte separation medium. 3
Isolation－of－human－prostatic－smooth－muscle－cells
Human prostate tissues1.Human hyperplastic prostates were obtained during surgery from four men through transurethral resection of the prostate. 2.All
Establishment－of－Stable－Transfectant－of－CHO－Lec－Cells
Purpose and BackgroundsCHO lec 3.2.8.1 cellsCHO Lec 3.2.8.1 cells have four independent mutations in the N- and O- glycosylation pathways (Stanley, 19
FACS－Analysis－Using－Peripheral－Blood－Cells
FACS Analysis Using Peripheral Blood CellsCollect blood (75 microliters) into 1ml PBS containing 5 microM EDTA (10 microliters of 0.5 M stock) and mix
Culturing－Rat－Fetal－Neural－Stem－Cells
实验概要Rat neural stem cells (NSCs) serve as a well-established model for investigating human brain development, disease processes, and treatment stra
Eccles:Protein－Lysates－from－Cells－in－Culture
Cell Lysis Buffer5mL 0.1M Tris HCl pH 8 (10mM)0.44g NaCl (150mM)0.02g EDTA (1mM)0.5mL nonidet P40 (1% w/v)0.05g SDS (0.1% w/v)Make up to 50mL with MQH
Isolation－of－human－uterine－smooth－muscle－cells
1. Approval to utilize human tissue was given by Hospital's Committee on Research Involving Human Subjects. 2. Human myometrial and leiomyoma tiss
Isolation－and－characterization－of－rat－glomerular－endothelial－cells
Glomerular endothelial cells (GECs) from the kidney are in close juxtaposition to other cell types, such as mesangial cells, and may respond to as
Isolation－of－human－corneal－endothelial－cells－(HCECs)
Isolation of HCECs1. The corneoscleral tissues were rinsed three times with primary cell culture system containing 50 mg/mL gentamicin and 1.25 mg
使用饲细胞(Feeder－Cells)制备
（1）取原代培养的人或动物胚胎成纤维细胞，90%汇合时制成细胞悬液，再按105细胞/毫升重新接种培养。（2）在细胞半汇合时，准备0.25μg/ml的丝裂霉素C，按2μg/106细胞的量加入到培养瓶中过夜；或用射线单次照射，剂量30～50戈瑞。（3）细胞经上述处理后，Hanks液漂洗两次、更换培养液、
Tissue－Culture－of－PtK1－cells
Please note: lately (2005 - present), we normally culture PtK1 cells in F-12 media, but they also grow in other types of media as described below.Mixi
Handling－Fresh－Hepatocytes－in－Suspension
实验概要Important—Please read!GIBCO® Fresh Hepatocytes are shipped in cold preservation medium designed to keep cells viable at 4°C. This shipping mediu