Magnification of the system: 3x, 5x, 10x, 20x Excitation (nm)
| Dichroic (nm)
| Emission (nm)
| Common Dyes
| 377/50 | 409 LP
| 438/24 | DAPI, Hoechst, Alexa Fluor 350, Aminomethylcoumarin, Lysosensor Blue, Marina Blue, Pacific Blue, BFP, EBFP | 438/24 | 458 LP
| 483/32 | AmCyan, CFP, ECFP, BOBO-1, SYTOX Blue
| 472/30 | 495 LP
| 520/35 | Cy2, CyQuant, DiO, YOYO-1, GFP, EGFP,
| 482/35 | 506 LP
| 536/40 | Carboxyfluorescein. FITC, Alexa Fluor 488, Calcein, Fluo-4, Mitotracker Green, Oregon Green, Rhodamine 110, rsGFP
| 525/40 | 555 LP
| 585/40 | Rhodamine, TMR, TRITC, Alexa Fluor 532, 5-TAMRA, Dil, POPO-3, PO-PRO-3, | 531/40 | 562 LP
| 593/40 | Alexa Fluor 546, Alexa Fluor 555, Calcium Orange, Cy3, LysoTracker Yellow, MitoTracker Orange, Phycoerythrin (PE), dsRed | 562/40 | 593 LP
| 624/40 | Propidium Iodide (PI), Cy3.5, Alexa Fluor 568, Alexa Fluor 594, Calcium Crimson, MitoTracker Red, Texas Red, HcRed
| 628/40 | 660 LP
| 692/25 | Cy5, Alexa Fluor 647, Alexa Fluor 660, APC (Allophycocyanin), BODIPY 650/665, DiD, SYTO Red 60, 62, 63, TOTO-3
|
Do note that due to the flexible combination of the filters, special dyes such as QDots are not an issue. Please check with us if you have any queries | SOP Start-up 1. Turn on the LEAP instrument by switching on the main power button. 2. Turn on the LEAP computer. 3. Launch the LEAP application using the “LEAP” icon, located on the desktop. The program opens and the initial startup screen, called Power System Warm-Up Status 4. Tick on the Excitation Lamp and wait for it to be warmed up. Pre-operation 1. Select magnification 2. Select the Laser to be used 3. Set the Laser power and spot size (with guidance from the Excel Spreadsheet on the desktop) Operation 1. Load the plate 2. Select the plate format 3. Load the Cell Purification Protocol 4. Load the channel settings (Example: channel 1 DAPI, Channel 2 GFP) 5. Optimise the exposure time and segmentation settings 6. Select the wells to be used 7. Run the protocol 8. Process the well for the Laser to fire 9. Run protocol again to verify selection efficiency Shut-down 1. Remove your plate 2. Close the LEAP software 3. Shutdown the computer 4. Power off the main system Sample preparation Please read attached phototherm reagent file |