Who we are
Where we are
The confocal microscopy laboratory is located on the ground floor of the Integrated Biological Complex at 39 Volturno Street
.
What we do
Confocal microscopy allows imaging by eliminating out-of-focus fluorescence interference on live or fixed preparations subjected to fluorescent marking. Production of the confocal image is achieved by illuminating the specimen with a beam of laser light that is sequentially focused on each individual spot on the preparation, either along the XY or Z plane. The acquisition of multiple confocal planes of the analyzed sample then allows its subsequent three-dimensional reconstruction using specific software. Technological evolution in the field of confocal microscopy has enabled the development of increasingly high-performance confocal systems in terms of scanning speed, resolving power and detection sensitivity of fluorescent signals. In the Confocal Microscopy Laboratory, afferent to the Department of Medicine of our University, a state-of-the-art confocal system supplied by the company Leica Microsystems has been installed.
Main equipment provided
The LEICA Stellaris 5 confocal system is equipped as follows:
-microscope DMi8 inverted revolver equipped with 4 objectives (10X/0.3; 20X/0.7; 40X/1.3 oil; 63X/1.4 oil) and motorized traineeship on XYZ axis
-White laser and AOBS dichroic filter programmable for simultaneous separation of up to 8 laser lines in excitation and between 470 nm and 670 nm. The Laser module is complemented by a 405 nm source for excitation in the UV.
-Fluorescence detection system is equipped with 3 independent "HyDS" spectral photo detectors with high quantum efficiency and configurable for acquisitions in analog mode, photon counting or λ-scan.
-"Lightning" acquisition mode for super resolution up to 120nm on XY-axes and 320nm on Z-axis
-top traineeship incubator at CO2 for time-lapse morpho-functional studies on live cell cultures
-double license of LasX software for 3D processing, Co-localization, FRAP, FRET and allow analysis of previous acquisitions in offline mode, on hardware platform independent from the confocal system.
The confocal microscopy office is completed with the recent upgrade of a Zeiss Axiovert 200 stand used for fluorescence widefield use. The system allows the acquisition of digitized images of fixed or viable biological preparations through the integration of a CO2incubator. The microscope is intended to provide an evaluation service prior to a thorough confocal analysis of the specimen under examination as well as to provide an alternative solution in case of particularly weak fluorescent signals that cannot be detected by a confocal approach.
The Zeiss Axiovert 200 fluorescence widefield microscopy system is configured as follows:
inverted revolver stand equipped with 5 objectives (10X/0.3; 20X/0.7; 40X/1.3 oil; 63X/1.4 oil; 100X/1.4 oil)
-3 sets of filters in excitation (BP 365/12; BP 450-490; BP 546/12)
-digital color camera Axiocam 305 controlled by ZEN software
-possibility of multichannel acquisitions
How to book analysis sessions
-For internal users, by contacting any of the authorized personnel available to use the microscope pertaining to your Department
-For University Departments that have not identified any contact person, by contacting any of the authorized personnel available to perform the measurements
-For external users, addressing any of the authorized personnel available to carry out the measurements
To schedule analysis sessions, consult the schedule for use of the instrumentation of interest by logging on to the following link:
LINK
Confocal Microscopy office fee schedule:
| Type of analysis | INTERNAL USER | EXTERNAL USER |
| New Acquisition | 60 euro/4h* | 200 euro/4h |
| Extra time** | 10 euro/h | 30 euro/h |
| Post processing | 10 euro/h | 30 euros/h |
* means minimum bookable machine hours, corresponding to half a day's office time
** means machine time beyond 8h of office time
Widefield Observation Tariff Schedule (ZEISS AXIOVERT 200)
| Type of analysis | INTERNAL USER | EXTERNAL USER |
| New Acquisition | 10 euros/h | 30 euros/h |
IMPORTANT!! Reservations may be cancelled:
- for FIXED samples up to the third working day prior to the requested date
for LIVE samples up to 24 hours prior to the requested date
Non-use will result, in the absence of timely cancellation, in the payment of the office
.
