细胞技术

AimIn cases of gross contamination the naked eye may identify the presence of bacteria and fungi. However it is necessary to detect low-level infections by incubation of cell cultures and/or their pro ...

AimDetection of mycoplasma by culture is the reference method of detection and has a theoretical level of detection of 1 colony-forming unit (cfu). However there are some strains of mycoplasma that ar ...

AimDNA staining methods such as Hoechst staining techniques are quick with results available within 24 hours which compares favorably with 4 weeks for detection by culture. However the staining of cul ...

The onion root tip and the whitefish blastula remain as the standard introduction to the study of mitosis. The onion has easily observable chromosomes and the whitefish has one of the clearest views o ...

PROPIDIUM IODIDE: The most commonly used dye for DNA content/cell cycle analysis is PROPIDIUM IODIDE (PI). It can be used to stain whole cells or isolated nuclei. The PI intercalates into the major gr ...

Solutions70% ethanol ribonuclease (100 µg/ml DNase free Sigma) propidium iodide ( 50 µg/ml in PBS)ProcedureHarvest cells. Spin at 1200 rpm for 5 minutes. Discard supernatant; add 1 ml ' fridge cold' 7 ...

1. Harvest cells- wash 2X in PBS to get rid of serum proteins. 1200rpm 5 min2. Resuspend pellet (up to 3x106 cells) in 1.2 ml PBS (Ca and Mg free).3. Add 3.0 ml ice cold 95% EtOH dropwise while vortex ...

Fixation1) Collect 2 X 106 cells.2) Pellet cells by spinning at 1000 rpm 4°C for 5 minutes.3) Resuspend cell pellet in 1 ml of cold PBS.4) Fix cells by adding 4 ml of -20°C absolute ethanol.5) Store c ...

MaterialsP.I. Solution: 4 mM Na3Citrate (0.118 g/100 mL)30 U/mL RNAseI (43 mg/100 mL)0.1% Triton-X100 (0.1mL/100 mL)50 µg/mL propidium iodide (5 mg/100 mL)ProcedureHarvest cells: Rinse with a subconfl ...

MATERIALS:1. 1 X PBS (PBSAz 1 X PBS e.g. Irvine Scientific CA containing 2% newborn calf serum and 0.1% sodium azide)2. 7-Amino-actinomycin D (7-AAD e.g. Calbiochem CA)3. Human AB serum heat-inactiv ...

Viable cells are cells that when allowed to continue beyond the timepoint of examination will stay alive. Besides live and healthy cells cells in early stages of apoptosis may be considered viable as ...

Materials: Hoechst 33258 (Sigma B-2883). stock: 10 mg/ml in dH20 (40) working dilution: 500µg/ml (50µl stock + 950µl PBS). 7-Amino-actinomycin (Sigma A-9400) stock: 200µg/ml in dH20 (40) 12x75mm snap ...

Currently the Moflo instrument (Dakocytomation) is used to sort cells at the AECOM FACS facility.Sorting is performed by the person in charge at the facility.You need to book an appointment in advance ...

BackgroundThis fixation method is good for cells labelled by fluorochrome-conjugated antibodies to membrane antigens. It will stabilize the light scatter and labelling for up to to a week in most inst ...

METHOD 1:Formaldehyde preservative � 2% formaldehyde solution in protein-free phosphate-buffered saline (PBS).Prepare as follows:Add 2 g paraformaldehyde powder (e.g. Sigma St. Louis MO) to 100 ml o ...

DescriptionProcedure for CD3/ CD4 /CD8 T cell subset counts using CD3-FITC / CD4-PE / CD8-PECy5 antibody Procedure1. One 5 ml round bottom tube (Falcon) is taken per blood sample to be analyzed. The t ...

Use this Protocol for Directly or Indirectly staining cells for Flow Cytometric Analysis 1) Dilute cells to 5x10^6 cells/mL2) Aliquot 100uL of cells per per tube (5x10^5 cells total)3) Add 20uL blocki ...

ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as a control.1 X PBS2% Buffered formaldehyde solution (see recipe)70% Ethan ...

ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as control.Hoechst 33342 stock solution (1mg/ml) (see recipe)12 X 75 mm cul ...

DescriptionProtocol for intracytoplasmic staining of cytokines for FACS analysis Procedure1) Prepare spleen lymph node or T cell clone cells as single cell suspension using erythrocyte lysis (water) o ...