LiveCellImagingofYeast
Live Cell Imaging of YeastDaniel R. Rines, Dominik Thomann, Jonas F. Dorn, Paul Goodwin and Peter K. SorgerINTRODUCTIONThe development of cloning vectors for green fluorescent protein (GFP) and the simplicity of yeast reverse genetics allow straightforward labeling of yeast proteins in living cells. Budding and fission yeast are therefore attractive organisms in which to study dynamic cellular processes such as growt......阅读全文
Live－Cell－Imaging－of－Yeast
Live Cell Imaging of YeastDaniel R. Rines, Dominik Thomann, Jonas F. Dorn, Paul Goodwin and Peter K. SorgerINTRODUCTIONThe development of cloning vect
Live－imaging－with－Drosophila－tissue－culture－cells2
Materials & ReagentsDrosophila Schneider S2 cellsSchneiders Medium (GIBCO/Invitrogen), 10% fetal calf serum, Antibiotics (Sigma A5955)Depression slide
Live－imaging－with－Drosophila－tissue－culture－cells1
IntroductionLive imaging provides an important complementation to the "snapshot" view obtained in fixed tissue by immunofluorescence. It allows follow
Cell－Counting/－LiveDead－Discrimination
This is a microscopy based application for definitive discrimination of live and dead cells. Trypan Blue exclusion notoriously over estimates the numb
Yeast－Cell－Cycle－by－Flow－Cytometry
ReagentsCold absolute ethanol.0.5 M Na citrate stock (filtered), 50mM diluted stock.10 mg/ml RNase A (Boil 10 mins, cool, filter and store at -20°C).4
LIVE/DEAD®－Fixable－Dead－Cell－Stain－Kits
实验概要The LIVE/DEAD® Fixable Dead Cell Stain Kits use a novel method to evaluate the viability of mammalian cells by flow cytometry. These assays are
LIVE/DEAD®－Fixable－Dead－Cell－Stain－Kits
实验概要The LIVE/DEAD® Fixable Dead Cell Stain Kits use a novel method to evaluate the viability of mammalian cells by flow cytometry. These assays are
Small－Scale－Yeast－Whole－Cell－Extract－for－IP
Grow 100 ml yeast cells in desired media overnight to an A600 of ~1.0 (0.6 to 1.2 works well). For growth in minimal media, 1 ml of a saturated overni
线粒体荧光探针大全：TMRM，Mitotracker，JC1（3）
Mitochondrion-Selective Rhodamines and RosaminesRhodamine 123Rhodamine 123 (R302; FluoroPure Grade, R22420; Figure 12.22) is a cell-permeant, cationic
SingleVirus－Tracking－in－Live－Cells
Single-Virus Tracking in Live CellsMichael J. Rust, Melike Lakadamyali, Boerries Brandenburg and Xiaowei Zhuang INTRODUCTIONReal-time, live-cell imagi
线粒体荧光探针大全：TMRM，Mitotracker，JC1（4）
Nonyl Acridine OrangeNonyl acridine orange (A1372) is well retained in the mitochondria of live HeLa cells for up to 10 days, making it a useful probe
Yeast－Media
YEPD (non-selection)-1% yeast extract-2% peptone-1.5% agar (if needed for plates)After autoclaving add glucose to 2% by adding 100 ml of 20% solution
In－Vivo－Luciferin－Imaging－Procedure
Mice are injected by an intraperitoneal route with a Luciferin solution (15 mg/mL or 30 mg/kg, in PBS, dose of 150 mg/kg) that is allowed to distribut
Fluidigm公司微液流芯片在单细胞研究中的应用（一）
Nature杂志在2009年5月7日的主页文章中大篇幅地介绍了美国Fluidigm公司的微液流芯片在单细胞表达中的应用。A closer look at the single cellMegan ScudellariNature Reports Stem CellsPublished online:
免疫荧光
Immunofluorescence Technique (Spector Lab)protocol for immunofluorescence on cells Immunofluorescence Protocol (Walter Steffen)Methanol fixationForma
从成体昆虫中分离活细菌的方法
Isolation of live bacteria from adult insectsHoracio M Frydman , hfrydman@princeton.edu, Associate ResearcherRelated Journal & Article InformationManu
从成体昆虫中分离活细菌的方法
Isolation of live bacteria from adult insectsHoracio M Frydman , hfrydman@princeton.edu, Associate ResearcherRelated Journal & Article InformationManu
Dropout－plates－for－yeast
Materials(Solutions are all available from the media room)200ml bottle of 2x SD200ml bottle of 4% agar -- make sure to sign it out40% glucoseCSM minus
Fast－Yeast－Transformation
Protocol: Fast yeast transformationAdd 50 µl carrier DNA to a 1.5 ml tube.scrap cells from plate and add to the carrier DNA.Add in the following order
Yeast－DNA－Prep
Protocolgrow up yeast culture to appropriate density (near saturation)spin 1.5 mls of culture for 1 min in microfuge and aspirate off supernatantresus
yeast:Assaying－mating
SetupYou have yeast strains that are deficient in mating (eg Ste12 knockouts) and would like to test whether transforming them with a plasmid that con
Yeast－Lysates－for－Westerns
Cells are grown for 2-3 days as 1.5ml prep. under selection for the plasmid of interest. Spin cells down 2.6K for 5min.Resuspend in 1ml 0.25m NaOH/1%
Preserving－yeast－cultures
Short term storageYeast cultures are stable for 1-2 weeks when refrigerated. Petri dishes should be sealed or in plastic bags.Medium term storageYeast
Modified－Yeast－Transformation
Inoculate cells from an overnight culture into 50 ml YEPD and incubate at 30°C with shaking. Typically, add 0.1 to 0.2 ml saturated culture in the eve
Yeast－Nuclei－Isolation
This method gives yeast nuclei which look nearly purified microscopically. Nuclei isolated in this way do not give active transcription extracts when
In－Vivo－Imaging－of－Far2
In vivo bio-imaging Mice were anesthetized and placed in a custom-made bed, which allowed stable and reproducible imaging of the legs. In vivo scann
In－Vivo－Imaging－of－Far1
In Vivo Imaging of Far-red Fluorescent Proteins after DNA Electrotransfer to Muscle TissueDNA electrotransfer to muscle tissue yields long-term, high
In－Vivo－Imaging－of－Far3
To determine the minimum dose of Katushka plasmid needed to give detectable fluorescent intensity, we decreased the amount of pTurboFP635 to 0.5 and 1
Heterogeneity－of－SingleCell－Gene－Expression－Across－Phenotypically（一）
Introduction Multi-cellular populations are fundamentally driven by the collective properties of individual cells. However, our understanding of ge
LIVE/DEAD®－Violet－Viability/Vitality－Kit
实验概要The LIVE/DEAD® Violet Viability/Vitality Kit provides a two-color fluorescence cell viability and vitality assay that is based on the simultaneo