Research | Optical microscopy laboratories

Activities

The optical microscopy laboratories are equipped with two confocal microscopes and three widefield optical microscopes. The application of laser scanning confocal microscopy enables high-resolution, high-contrast imaging with optical sectioning, allowing reconstruction of precise 3D images of the specimen. The Zeiss Axioplan 2 imaging stand and the Leica DMI6000 B, on the other hand, are for fluorescence widefield and brightfield use. The systems enable digitized imaging of fixed preparations and real-time imaging of live cells, as in live-cell imaging and time-lapse imaging, respectively, which are useful for monitoring intercellular interactions and analyzing the dynamics of specific subcellular processes. A Zeiss Axiovert 5 is also available, which is used to acquire digitized images of cell cultures. The Leica CM1950 cryostat for sectioning histological preparations completes the equipment.
The microscopy laboratory staff is responsible for managing the advanced bioimaging equipment, providing routine maintenance, instrument upgrades, training and technical support to users.

Equipment

List of instrumentation provided

Confocal microscopes:

• Leica TCS SP8 confocal microscope

• Zeiss LSM 510 Meta confocal microscope

Widefield optical microscope:

• Inverted fluorescence optical microscope with thermostatted chamber and CO2 control aimed for use in in-vivo imaging Leica DMI6000 B

• Zeiss Axioplan 2 Straight Fluorescence Microscope Imaging

• Zeiss Axiovert 5 inverted microscope

Leica CM1950 Cryostat

Instrumentation description.

Leica TCS SP8 confocal microscope

The Leica TCS SP8 is a confocal fluorescence microscopy system that provides high-resolution images and three-dimensional reconstructions by optical sectioning. The system is based on a Leica DMi8 inverted microscope with motorized stage equipped with HC PL APO CS2 20x/0.75 NA Dry, HC PL APO CS2 40x/1.30 NA oil, HC PL APO CS2 63x/1.40 NA oil objectives and four laser excitation sources: Diode 405, Argon, DPSS 561 and finally HeNe 633nm. The acquisition system uses four conventional photomultipliers (PMTs) and a PMT Trans acquisition system for the bright field.

Zeiss LSM 510 Meta confocal microscope

The Zeiss LSM 510 Meta microscope is a confocal fluorescence microscopy system that provides high-resolution images and three-dimensional reconstructions by optical sectioning. The system is based on a Zeiss Axiovert 200M inverted microscope equipped with Plan-Neo 10X /0.3 NA, Plan-Apo 20X/ 0.50 NA, EC Plan-Neo 40X/ 1.30 NA Oil Dic, C-Apo 40X/1.2 NA W, Plan-Apo 63X / 1.4 NA Oil Dic and Plan-Apo 150X/1.35 NA Dic Gly objectives. It is equipped with four laser excitation sources: Argon 458/477/488 / 514nm, HeNe 543nm, HeNe 633nm and finally Diode 405 nm.

Fluorescence optical microscope with thermostatted chamber and CO2 control for in-vivo imaging Leica DMI6000 B

The DMI6000 B inverted optical microscope with motorized stage is equipped with HC PL Fluotar 10x/0.30 NA dry, HC PL Fluotar L 20x/0.40 NA dry, HC PL Fluotar L 40x/0.60 NA dry, HC PL Fluotar L 63x/0.70 NA dry and HCX PL Fluotar 63x/1.25 NA oil objectives and two cameras (DFC7000T and DFC350FXR2) that enable conventional and bright-field fluorescence imaging. The equipment allows time-lapse and 3D acquisitions and is implemented with an Okolab incubation system that allows observation of live biological specimens, using a temperature and CO2 control module.

Zeiss Axioplan 2 imaging fluorescence microscope

The Axioplan 2 imaging straight optical microscope is equipped with A-Plan 2.5x/0.06 NA, Plan-Neofluar 10x/o.30 NA Ph1, Plan-Neofluar 20x/0.50 NA, Plan-Neofluar 40x/1.30 NA oil, Plan-Neofluar 63x/1.25 NA oil, Plan-Neofluar 100x/1.3 NA oil objectives. Allows observation and acquisition of digitized images in both brightfield and fluorescence.

Zeiss Axiovert 5 optical microscope

The Axiovert 5 optical microscope enables bright-field imaging of cellular samples using Axiocam 208 color camera. It is equipped with the following objectives: LD A-Plan 5x/0.15 NA Ph1, LD A-Plan 10x/0.25 NA Ph1, LD A-Plan 20x/0.35 NA Ph1 and LD A-Plan 40x/0.55 NA Ph1.

Leica CM1950 Cryostat

The Leica CM1950 cryostat enables sectioning of histological preparations included in O.C.T. It has a temperature adjustment range from 0 °C to -35 °C, and the section thickness range is from 1 to 100 μm.

Price list

Available at the following link:
https://www.difarma.unisa.it/uploads/rescue/472/2261/tariffario-difarma.pdf

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