Chapter 1: Introducing Polaris - Fluidigm Polaris User Manual

Chapter 1: Introducing Polaris

The Polaris™ system is a flexible microfluidics platform that allows you to perform
single-cell gene expression analysis. Under automated control, Polaris can image,
select, capture, culture, and dose single cells individually.
You can perform multi-step molecular biology protocols on the captured, single cells
and export the reaction products for further analysis. Thus, in situ single-cell analysis
can be combined with the study of alterations in the entire transcriptome or genome.
The Polaris instrument uses the Polaris Single-Cell Dosing mRNA Seq IFC. This IFC
(integrated fluidic circuit) captures 48 cells varying in size from 5 to 25 microns and is
capable of measuring responses to environmental changes by allowing treatment
and culturing of cells for up to 24 hours.
Follow the procedures in the document Polaris mRNA Seq Protocol (PN 101-0082),
which shows you how to select single cells; dose, feed, and/or treat them under
controlled environmental conditions; image targeted cells with fluorophore stains
(for example, to ensure cell viability); harvest their cDNA; and generate a cDNA
library for sequencing. You can see an example of the Results step for a Dose
Response experiment in Run: View the Results on page 35.
The following table is an example of fluorophores that can be used on Polaris:
Table 1. Examples of cell-staining dyes
Channel Name
(FAM)
(VIC)
(Cy5)
(ZOMBIE)†
* Excitation values are center wavelength/pass band (≥90%).
† Zombie Yellow is only for post section and treatment staining.
6
Excitation * Emission
475/40 525/25
530/20 570/30
632/28 700/30
438/28 570/30
Recommended Stains
Alexa Fluor ® 488 (selection marker)
CellTracker™ Orange CMRA Dye (universal marker)
Alexa Fluor 647 (selection marker)
Zombie Yellow™ (dead cell dye)
Polaris: User Guide