We at the Olympus’ Advalytix team believe that single cell molecular analysis should not be unreliable and cumbersome. Single cell analysis should be Simple, Seamless, and Sensitive®, a concept that we call Zellvergnügen® (German for the “pleasure of cells”). We have developed workflows to guide researchers through single cell analysis, no matter whether single cells are captured via flow sorting, micromanipulation, or laser capture microscopy, or whether downstream analysis involves qPCR, PAGE, CE, or sequencing see video. Step 1: Isolation & deposition Three easy ways to isolate and deposit single cells on AmpliGrid slides: Take the guess work out of cell placement! Visual QC result: verify correct cell placement
FACS™ see video
• For liquid samples with cells in suspension
• High throughput cell sorting/deposition
• Placement accuracy superior to sorting into MTPs
Laser Capture Microdissection (LCM)
• Isolate cells from tissue (pathology, oncology, neurology)
• Direct deposit of cells with PALM system
Micromanipulation see video
• For standard workflows using capillaries for cell picking and transfer, e.g. IVF
• Automation available
Step 2: Quality control see video
Stain your cells (e.g. fluorescent DNA stain) prior to deposition. Then check sorted slide for accurate cell placement with a fluorescent microscope (US and EU customers). This process can also be automated.
Step 3: Amplification see video
AmpliSpeed Slide Cycler for AmpliGrid slide
For amplification, simply add 1µL of mastermix and cover with 5µL of cover oil. Hydrophobic areas confine mastermix and oil to their designated areas – no contamination, no mess.
• Sensitive amplification of DNA, mRNA, mtDNA
• Enhanced sensitivity with DNA extraction kit
• Manual or automated pipetting (see video on Youtube)
• 1uL assay volume
• Pre-amplification for qPCR
Step 4: Analysis see video
Gel Analysis/ Page
Determine presence/absence of up to 5 genes/transcripts, or isolate fragments for subsequent sequencing. Let us design your multiplex assay.
Capillary Electrophoresis
Detect up to 25 genes/ transcripts from a multiplex assay. Get semi-quantitative results by mapping the area under the peaks into low, medium and high expressers.
Sequencing
Combine RT-PCR/PCR with sequencing on AmpliGrid
qPCR
Add AmpliGrid’s convenient single cell workflow and sensitivity with to established qPCR data analysis.
BD, FACS and FACSAria are registered trademarks of Becton, Dickinson and Company, USA
