PerkinElmer Opera Phenix - Specifications

General information

Microscope type: Spinning disk confocal and widefield
Purpose: High-throughput and high-content imaging of multi-well plates will full automation
FPbase link: https://www.fpbase.org/microscope/rBKJsuQSCGgSnZuxcGRYSh/

Equipment information

Location: JSCBB C350C

Serial number: 1400L18161
CU Tag: UCB 214288 RC16

Sample types

Fixed Cells	Yes
Live Cells	Yes

Software

• Harmony v5.6

Environmental control

• CO₂, Humidity, Temperature

Laser lines

• 375 nm or 425 nm
• 488 nm
• 561 nm
• 640 nm

Filters

Emission				Example fluorophores
Center (nm)	Width (nm)	Min (nm)	Max (nm)
457.5	45	435	480	DAPI, BFP, CellMask Blue
472.5	15	465	480	CFP
475	80	435	515	CFP/YFP FRET: Acceptor Only
492.5	115	435	550	DAPI, BFP, CFP, CellMask Blue
497.5	65	465	530	DAPI, BFP, CFP, CellMask Blue
515	30	530	500	Alexa 488, GFP
525	50	500	550	GFP, Alexa 488, FITC, CellTracker Green, YFP
532.5	35	515	550	YFP
583.5	25	571	596	Alexa 568, Cy3, TRITC, dsRed
600	60	570	630	TRITC, Texas Red, dsRed, Alexa 568, Alexa 594, mCherry
617.5	25	605	630	Alexa 594, mCherry
665	30	650	680	Alexa 647, Cy5, CellMask Deep Red
705	30	690	720	Cy5.5, Draq5
705	110	650	760	Cy5, Alexa 647, Draq5

Please note that these emission filters are tied to specific laser lines/cameras, indicated by color above.

• 375 nm or 425 nm laser → 457.5 nm, 472.5 nm, or 475 nm filter → Camera 1
• 488 nm laser → 492.5 nm, 497.5 nm, 515 nm, 525 nm, 532.5 nm filter → Camera 2
• 561 nm laser → 583.5 nm, 600 nm, or 617.5 nm filter → Camera 3
• 640 nm laser → 665 nm or either 705 nm filter → Camera 4

Objectives

Magnification	NA	Immersion medium	Field of view (mm)	Effective xy resolution (µm), binning = 1*	Effective xy resolution (µm), binning = 2*	Recommended minimum distance between planes (µm)**
5X	0.16	Air	2.58 x 2.58	2.6	5.3	26.5
10X	0.3	Air	1.29 x 1.29	1.3	2.6	7.4
20X	0.8	Air	0.65 x 0.65	0.66	1.32	1.2
20X	1.0	Water	0.65 x 0.65	0.66	1.32	0.8
40X	1.1	Water	0.32 x 0.32	0.33	0.66	0.5

*Calculated at 520 nm. Optical resolution = 0.61 ʎ_emission / NA.

**Calculated as "Depth of focus / 2.2" (according to Nyquist's theorem). Please note that using smaller distances will not improve the results.

Detectors

• Andor Zyla x4 sCMOS camera
• 16-bit camera: pixel intensity values from 0 - 65,535
• Camera ROI: 2160 x 2160 pixels

Instrument diagram