Type of device

High-Content Imager / Confocal Scanning Microscopy / Live-Cell Imaging
Operetta CLS by PerkinElmer (Revity)

Short description

The device is flexible for different imaging situations that require quantification:

Well plate plate types and slides
Automated re-imaging, e.g., re-image cells with 10% largest nuclei at higher magnification
Widefield and spinning-disk confocal imaging
Live-cell imaging (37°C, CO₂ variable) over time also with water-objectives (autom. refill)

Instrument parameters

Technical specifications:	sCMOS camera Motorized stage Confocal Disk Transmitted light and digital phase contrast Ratiometric FRET 3D-imaging and live cell imaging Fluorescence, fluorescence transmittance, transmission
Objectives:	3 air and 3 water-emmersion objectives permanently installed All objectives in the table below
Laser:	8 LEDs
Software:	Harmony 4.9

Objectivs
Objective	NA	Working distance [mm]	Corrected for plate thickness [mm]	Field of view [mm]	Dept of focus [µm]
1.25x Air	0.03	4	0.17	10.33 x 10.33	1501.0
5x Air	0.16	12.1	0.17	2.58 x 2.58	58.2
10x Air	0.3	5.2	0.17	1.29 x 1.29	16.2
20x Air	0.4	8.28	0 - 1.5	0.65 x 0.65	8.0
20x Water	1.0	1.7	0.17	0.65 x 0.65	1.8
40x Air	0.6	3.28	0 - 1.5	0.32 x 0.32	3.3
40x Water	1.1	0.62	0.17	0.32 x 0.32	1.2
63x Water	1.15	0.6	0.17	0.21 x 0.21	1.0

Laser: 8 LEDs
Wavelength	Power	Excitation Filter	Classification
365 nm (UV)	130 mW	355-385 nm	Risk Group 3
405 nm	230 mW	390-420 nm	Risk Group 3
450 nm	230 mW	435-460 nm	Risk Group 3
475 nm	110 mW	460-490 nm	Risk Group 2
505 nm	100 mW	490-515 nm	Risk Group 2
550 nm	170 mW	530-560 nm	Risk Group 2
630 nm	120 mW	615-645 nm	Risk Group 2
660 nm	250 mW	650-675 nm	Risk Group 3

Autofocus Laser
Wavelength	Laser Power	Descrption
785+/-10 nm	12 mW	sCMOS camera Laser class 3B

Transmission Light Source
Wavelength	Laser Power	Descrption
740 nm	80 mW	LED for Brightfiled and Digital Phase Contrast measurements Integrated in sample lid

Methods & Expertise

Many different imaging assays and quantifications, e.g., migration assay, 3D spheroid growth tracking, organoid imaging and three-dimensional object segmentation and quantification, zebrafish larvae imaging, actin length quantification, scratch assays, single-cell sorting verification (single cell per well) > all in 96-well plates or similar; whole tissue section imaging on up to four slides, prescan-rescan assays for automated re-imaging of specific sub-populations within cell lines and primary cells

Reference publications

Over 10 publications together with University (Medicine) Greifswald, e.g., on human organoids (DOI: 10.1007/s00018-022-04355-6 Fig. 4e)
Over 30 publications in house, e.g., on high-throughput micronucleus analysis (DOI: 10.3390/nano12244463 Fig. 1), actin protrusion quantification (DOI: 10.1038/s41598-018-37169-3 Fig. 5), and whole-tissue imaging and quantification (DOI: 10.1080/2162402X.2024.2425124 Fig. 1-3)

Affiliation, Contact & Terms of use

Affiliation:	XXXX
Location:	XXXX
Contact person:	XXXX
Terms of use:	XXXX
Biosafety level:	None/S2