UWFlowLSRMap032718
Optical Bench Map of BD LSR Flow Cytometer with common fluorochromes and optical filter specs
File: UWFlowLSRMap.pdfFlow Cytometry Lab
Rainbow Bead Standardization – Attune
Technical Note describing method and steps for incorporating a bead standard into flow cytometry assays acquired on the Attune flow cytometer.
File: Rainbow-Standard-Tech-Note-Attune.pdfFlow Server Connect Mac
Instructions for connecting to UWCCC Flow Lab data server using FILR on a Mac
File: Mac-Filr-Access-to-Flow-Data-Server.pdfFlow Server Connect PC
Instructions for connecting to UWCCC Flow Lab data server using FILR on PC
File: PC-Filr-Access-to-Flow-Data-Server.pdfRegaining Keyboard Control After Diva Locks it Out
Periodically, Diva will stop accepting input from the keyboard. If the mouse is working but you cannot type to annotate your data, try the method explained the document to regain control.
Updated 9/18/17
Biosafety guidelines for cell sorting
Cell sorting is an aerosol-generating procedure, with greatly increases the risk of airway transmission of hazardous and infectious material. Aerosol incidents are not uncommon during normal sort procedures. These incidents can be caused by sample clogging and other disturbances that increase physical instability of the fragile single-cell stream.
Updated 9/18/17
Cell Cycle Analysis
The cell cycle profile of a sample can be determined by staining the DNA with a fluorescent dye and measuring its intensity. The dye stains DNA stoichiometrically, allowing differentiations of cells in G0/G1, S phase, and G2/M, as well as identification of aneuploid populations. A variety of staining protocols can be adapted for different sample types, but the general analysis remains the same.
Updated 9/18/17
Doublet Discrimination
Good sample preparation practices are critical for flow cytometry. Filtering removes large aggregates that may otherwise clog the cytometer, but doublets and small multiplets will pass throug hand remain in the sample. Each clump passes through the lasers and is interpreted as one event with the combined properties of all cells in the group. These composite events must be excluded during analysis.
Updated 9/18/17
Sort Sample Prep Guidelines
Cell viability, autofluorescence and cell aggregation may affect the overall quality of live cell sorting assays. Superior cell preparation is crucial and will result in better sort purity, yield, and post-sort cellular function and viability.
Updated 9/18/17
Single Cell Sorting for Transfected Cells
Cell sorting creates aerosols that may pose a risk to the operator and others in the sort lab. Cell sorting must be included in your lab’s Biological Safety Protocol and on file with the UW Office of Biological Safety.
Updated 9/18/17