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Immunofluorescence Staining & Flow Cytometry of Cell Surface Antigens
protocol
Protocol from eBioscience, Inc. ...
FACScan Startup and Shutdown Procedure
protocol
From JCSMR FACS Facility. This procedure is for FACScan machine. ...
General cell staining protocol for flow cytometry
protocol
From FACS core facility, University of Connecticut Health Center. ...
Cytometer FACScan/Calibur/LSR/LSR-2 Operation Training Guide
protocol
From FACS core facility at University of Connecticut Health Center. ...
Labelling of Nuclei with BrdU and PI for FACS
protocol
Protocol of Labelling of Nuclei with BrdU and PI for FACS. (Finkbeiner lab, UCSF). ...
96 well microplate intracellular staining for FACS analysis
new
protocol
Protocol of microplate intracellular staining for FACS analysis from Dr. Cattoretti lab, Columbia University. ...
Flow cytometry protocols
new
protocol
Flow cytometry protocols from University of Pennsylvania, school of dental medicine.
content:
Direct Surface Immunophenotyping
PI Staining for Cell Cycle Determination PLUS FITC-Surface Staining
Three Apoptosis Detection Techniques
Hoechst/7 ...
Cell cycle analysis with PI and BrdU
new
protocol
Cell Cycle Analysis Using Propidium Iodide & Bromodeoxyuridine by W. Roy Overton, Ph.D.
Reagent and detailed procedure included. ...
Commonly used Cytometry Apoptosis Detection Techniques
new
protocol
Hoechst and 7-AAD
H342 is DNA stain that binds preferentially to A-T base-pairs without the necessary of permeabilization step. However, it is necessary to set up incubation conditions for efficient internalization of the dye. Hoechst 33342 (Bisbenzi ...
Common Fluorescent Dyes in Flow Cytometry
review
An introduction on commonly used fluorescent dyes in FACS analysis (from ebioscience inc). ...
Compensation in Flow Cytometry Analyses
review
This method review is written by Mario Roederer. Compensation is probably the least understood process accompanying flow cytometric analyses. Perhaps this is because it is often described with the linear algebra elements needed for its computation, and many of ...
Purdue University Cytometry Laboratories Web Site (PUCL)
recommended
site
A popular site for cytometry researchers with many valuable resources on cytometry application. ...
Flow Cytometry on the Web
site
This is the flow cytometry WWW links list located at the Salk Institute CCMI, La Jolla, California. ...
Collection of cytometry protocols from Janis V. Giorgi Lab
site
List of protocols from the Janis V. Giorgi cytometry Lab:
Flow Cytometry Glossary
Method for EtOH fixation of cells for long-term storage and DNA staining
Minimizing Background Staining
Blocking FC-Receptor Binding
Monoclonal Antibody Staining Proc ...
FACS Sensitivity
troubleshooting
Could anyone tell me the limit of sensitivity of fluorescence activated cell
sorting? Is it possible to detect, for example,one labelled cell out of a
population of 1 million cells?
... ...
Click following links for original posts.
...
Optimizing Sorting Experiments
troubleshooting
This page listed several factors that need to be considered in order to prepare cells properly for a successful FACS. (TSRI Flow Cytometry Protocols) ...
Table of Fluorochromes
software
This is a table of some characteristics of fluorochromes useful for flow cytometry or fluorescence microscopy. Within groups, roughly in order of excitation wavelength (families excepted)
From JCSMR FACS lab. ...
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Last update:  09-May-2008 01:11 am
Related new papers and reviews
Flow acetone-staining technique: a highly efficient procedure for the simultaneous analysis of DNA content, cell morphology, and immunophenotype by flow cytometry. Cytometry A. 2008 Feb;73(2):168-74
Authors: Carbonari M, Mancaniello D, Tedesco T, Fiorilli M
The accurate determination of cell cycle, immunophenotypes and morphology at single-cell level is not fully achieved by current flow cytometry protocols. Acetone, a coagulant fixative/permealizing agent, is widely used in static cytometry, but is impractical in flow cytometry because of its shrinking effect. We sought for conditions of acetone treatment that could permit the simultaneous analysis of physical parameters, surface and intracellular immunostaining, and DNA content. We evaluated different experimental conditions (concentration, duration of fixation, temperature, presence of proteins) to test the capacity of acetone fixation/permeabilization to preserve cell physical parameters (forward and side scatters, FSC, and SSC) and immunophenotyping while allowing stoichiometric DNA staining. The commonly used ethanol fixation technique was used as reference method. To detect phenotypes and DNA content simultaneously, we employed 7-aminoactinomycin D (7-AAD) as "intercalating" dye for DNA in spite of, or just for, its controversial ability in stoichiometric DNA staining. Cells were resting peripheral blood monucleated cells (PBMCs), T- and B-cell blasts obtained by PBMCs stimulation, and the human cell lines Ramos and Shep. Acetone fixation, preserving both the recovery and the physical parameters of cells, is drastically influenced by temperature of treatment and is practicable only when the protocol is realized at 8 degrees C. Under this condition, acetone maintains the immunophenotypic fluorescences (realized before or after the fixation) better than ethanol. Stoichiometric DNA staining of acetone processed cells, the variation coefficients (CV) of frequency distributions of G1/G0 and G2/M phases, the modes ratio of these distributions and doublets generation are at least comparable to those obtained with ethanol treatment. The assay developed in the present study, that we called flow acetone-staining technique (FAST), accurately analyzes cell cycle, physical parameters and immunophenotypes in heterogenous cell populations, and thus provides a useful tool for cytomics.
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