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Useful Links

Educational and Technical Links:

Education

International Society for Advancement of Cytometry - ISAC
http://www.isac-net.org

New England Cytometry
http://newenglandcytometry.com

Purdue University Flow and Image Cytometry
http://www.cyto.purdue.edu

http://www.cyto.purdue.edu/class/

http://www.cyto.purdue.edu/flowcyt/educate.htm

http://www.cyto.purdue.edu/flowcyt/labinfo/labinfo.htm

Maeckerlab Weblog - Research and training materials for flow cytometry and immune monitoring.
http://maeckerlab.typepad.com

BD Biosciences Educational Programs Online

http://www.bdbiosciences.com/us/s/training/e-learning/e-learningcourses for beginners

http://static.bdbiosciences.com/training/diva6_course/index.html?_ga=2.221200155.225408873.1527001100-1298382617.1506082957 for Diva training

http://www.bdbiosciences.com/us/s/webinars/archives

Howard M. Shapiro, MD.: Practical Flow Cytometry, 4th Edition (free ebook)
https://www.beckman.com/resources/reading-material/ebooks/practical-flow-cytometry

Tools

BD Product Selection Tools

https://www.bdbiosciences.com/en-us/applications/research-applications/multicolor-flow-cytometry/product-selection-tools

BD Biosciences Spectral Viewer
https://www.bdbiosciences.com/en-us/applications/research-applications/multicolor-flow-cytometry/product-selection-tools/spectrum-viewer

Cytek Spectrum Viewer
https://cytekbio.com/blogs/resources/tagged/tools

Thermo Fisher Spectra Viewer
https://www.thermofisher.com/us/en/home/life-science/cell-analysis/labeling-chemistry/fluorescence-spectraviewer.html

FluoroFinder
Design Experiments for UMASS Flow Core's cytometers’ laser and filter configurations.
https://umassmed.fluorofinder.com

Live/Dead Staining
Dead cells bind antibody non-specifically and can exhibit a higher autofluorescense. Thus it is very important to exclude dead cells from your analysis - or sorted cells to improve the quality of your experiment. There are a wide array of options to choose from. They range from the classic dyes (PI, DAPI, Hoechst or 7AAD) to the newer fixable live/dead stain versions.

Below you will find a selection of companies providing these dyes as well as offering very good explanations on how those dyes work and give examples.

Thermo Fisher LIVE/DEAD Fixable Dead Cell Stains
LIVE/DEAD Fixable Blue stain - UV
LIVE/DEAD Fixable Violet stain - 405
LIVE/DEAD Fixable Aqua stain - 405
LIVE/DEAD Fixable Yellow stain - 405
LIVE/DEAD Fixable Green stain - 488
LIVE/DEAD Fixable Red stain - 488 or 561
LIVE/DEAD Fixable Far Red stain - 633
LIVE/DEAD Fixable Near-IR stain - 633

Biotium Live-or-Dye™ Fixable Viability Stains
Live-or-Dye Fixable Viability Stains
Live-or-Dye NucFix™ Red, a unique cell membrane impermeable dye that specifically stains the nuclei of dead cells, labeled covalently and does not transfer between cells after fixation.

Bio-Rad Live-Dead Exclusion Stains
VivaFix Fixable Viability Dyes
ReadiDrop 7-AAD
ReadiDrop PI
PureBlu DAPI
PureBlu Hoechst 33342

BioLegend Live-Dead Zombie Stains
Zombie Fixable Dyes
Helix NP NIR (ex/em - 640nm/660nm)
Draq7 (ex/em - 633nm/695nm)
PI (ex/em - 493nm/636nm)
7AAD (ex/em - 546nm/647nm)

Abcam live-and-dead-cell-assay
Live and Dead Cell Assay

Filtration recommendations
- Sterile 12x75 mm tubes with filter caps; by Corning - Fisher Cat. number 08-771-23
- Nitex Nylon Filtration Mesh order from: ELKO Filtering Co, LLC. (elkofiltering.com) Nylon mesh roll 37µ opening size, 40" wide, sold by the yard) Item#: 03-37/24

The fabric comes in yards. Use a OLFA rotary cutting mat or just scissors to cut it into strips and then 1cm² pieces and store them in a tissue-culture dish (if sterilization is needed, use a glass petri dish which can be autoclaved). Place a square on the top of the tube and carefully add a drop of sample to the fabric so that the fluid that can penetrate and is pulled through by surface tension down the inside of the tube. Concentrated samples may resist filtration and may need to be diluted. When the sample is flowing through, you can add the rest of the sample being sure not to go too fast. If the entire surface of the fabric gets wet where it’s touching the tube, flow will stop as the air pressure builds up inside the tube.

- Miltenyi MACS^® SmartStrainers #130-098-458