Sony/iCyt  Reflection
An iCyt Reflection is available to provide sorting capability in a BSL-2+ environment. This enables the sorting of viable infectious or potentially infectious materials. Human cells of unknown infectious state or cells with known BSL-2+ infectious agents may be sorted on the Reflection. Cells containing BSL-3 (or higher) level organisms are not permitted. Cells with BSL ratings lower than BSL-2 (e.g. mouse cells) may also be sorted using the Reflection facility. The Reflection has at least 12 parameter analysis/sorting capability - forward and side scatter and 10 colors of fluorescence using up to 6 laser excitation wavelengths. The iCyt Reflection as currently configured in our facility has 2 sorting modules (called HAPS - Highly Automated Parallel Sorting) which can operate simultaneously. Up to 2 additional HAPS could be added. Each HAPS can sort up to 4 populations simultaneously. This configuration permits many opportunities for new approaches to sorting. For example: both sort units could be doing the same sort on the same sample (i.e get done twice as fast); each sort unit could be doing a different sort on the same sample; each sort unit could be doing the same sort on different samples (sort the cells from your experimental mouse and your control mouse simultaneously); a high speed enrich sort could be done on one HAPS and the product of that sort moved to the other HAPS for high purity sort - this could be done in an overlapping way to conserve time - very useful for rare event sorting.

Only staff personnel are permitted into the sort room. However, investigators may view the data and participate in setting up the sort via a networked computer in Rm. 618. Investigators may view the data live during the sort from other locations (e.g. office or lab) if they have a computer with WinList 6.0 (Verity Software House). Consultation with the Facility Director as well as completing appropriate documentation (Biohazard Sorting Form) is required before using the Reflection

Instrument Overview

Location: 623 Mary Ellen Jones Bldg.
Laser Excitations: 355nm, 408nm, 488nm, 532nm, 594nm, and 635nm

• at least 11 colors of fluorescence
• Two simultaneous sort channels
• culture plate deposition - single/multiple cell
• 4-way sorting on each sort unit
• BSL-2+ sorting

Laser/Excitation - Possible Fluorochromes

UV/355nm excitation - Hoechst, Quantum Dots

violet/408nm - Pacific Blue, Pacific Orange, DAPI, Violet DyeCycle
Live/Dead violet, Quantum Dots

blue/488nm - FITC, Alexa 488, GFP, PerCP, PerCP-Cy5.5

green/532nm - PE, PE-TxRed, Alexa532, PE-Alexa610, PE-Cy5,
PE-Cy5.5, PE-Alexa700, PE-Alexa 750, PE-Cy7, PE-Alexa 647, DsRed

orange/594nm - mCherry, mRFP, max FP-Red, Alexa 594, Texas Red
red/640nm - APC, APC-Alexa700, APC-Cy7, APC-Alexa 750, Alexa-700, Alexa-647