Error Status code: 401
Location : UIC Room 5 (0B05), Bartolomeu Dias Wing
Manufacturer : Zeiss
Model : LSM 980
Year Installed: 2019
Optics : Confocal fluorescence and DIC
Detectors : 2 PMT + 1 GaAsP + 1 TLD (Transmitted light - brightfield) + Airyscan2
Keywords:
User guide: link to pdf
Confocal microscopy allows one to optically section a fluorescently labelled sample with superior contrast and even higher resolution by using a pinhole to reject light that originates above and below the focal plane. By collecting a series of image planes (“optical slices”) through the full thickness of a sample,it is possible to assemble a three-dimensional reconstruction of the entire sample.
The Zeiss LSM 980 is an inverted microscope equipped with a laser scanning head with a total of 5 detectors:
1x blue-green multialkali PMT
1x GaAsP PMT spectral detectors (tunable between 450-750 nm)
1x green-far-red multialkali PMT
The microscope is also equipped with a dark heating and athmospheric control chamber to allow live imaging of mammalian cells.
Images were acquired on an Zeiss LSM980 system, [using Airyscan [SR/Multiplex4Y or 8Y] mode] equipped with two PMT and one GaAsP, using a 63x 1.4NA Oil immersion objective. Serial sections were acquired every ** µms. with Zeiss’s ZEN Blue v3.0.
We thank the technical support of IGC's Advanced Imaging Facility, which is supported by Portuguese funding ref# PPBI-POCI-01-0145-FEDER-022122, co-financed by Lisboa Regional Operational Programme (Lisboa 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER) and Fundação para a Ciência e a Tecnologia (FCT, Portugal).
Laser |
Excitation line |
UV |
405 nm |
Blue |
488 nm |
Green |
561 nm |
Red |
639 nm |
Magnification |
NA1 |
Immersion |
Working distance (mm) |
FOV (mm) |
Theoretical resolution / @Nyquist |
Link |
|
2.5x |
0.085 |
- |
8.8 |
10 |
3.30 / 1.32 |
||
10x ![]() |
0.45 |
- |
2.0 |
2.5 |
0.623 / 0.249 |
||
20x ![]() |
0.8 |
- |
0.55 |
1.25 |
0.35 / 0.14 |
||
25x ![]() |
0.8 |
water, silicone oil, glycerine or oil |
0.57 |
0.72 |
0.35 / 0.14 |
||
40x ![]() |
1.1 |
water |
0.62 |
0.625 |
0.26 / 0.10 |
||
63x ![]() |
1.4 |
oil |
0.14 |
0.397 |
0.20 / 0.08 |
(1) NA - Numerical Aperture. For more information, follow this link.
Theoretical resolution for each objective, which might differ from the actual pixel size of acquired image, calculated as 0.51*550nm/NA. Resolution formula is taken from here.
Nyquist = max resolution with sampling at 2.5x the theoretical resolution; must be used for images to be deconvolved.
The pixel size in microns of your picture is saved automatically. Use Bio-formats to recognize it with ImageJ/Fiji (already installed in the Fiji package).
DO NOT TURN OFF THE COMPONENTS SWITCH, THE STAGE AND PIEZO CONTROLLER THAT ARE LOCATED ABOVE THE LASER BOX
Acquiring images in a confocal is a complex procedure, first time users are required to contact the UIC staff to discuss the use of confocal microscopes and fluorescent imaging in general. If you need assistance don’t forget to press “with assistance” at reservation time.
Zeiss LSM 980 manual is in construction.