Technical assistance in the acquisition of contocal microscopy images using microscopy equipment Leica TCSP5.
In 2013 this Facility has been granted with the ISO 9001 Certificate: IQNetES-0286-2009_2019-01-08.pdf
• Stereo Microscope Leica MZ16F with color camera Leica DFC490. Transmitted and fluorescence light.
• CCD Digital Camera Leica DFC 350 FX coupled with a light microscopy Zeiss Axioplan with phase contrast technique and incident light fluorescence.
• Confocal Laser Scanning Microscope (CLSM) LEICA TCS SP5 fitted with eight excitation lines (405, 457, 477, 488, 496, 514, 543, 633 nm) from UV, Ar and HeNe lasers and AOTF (Acousto-Optical Tunable Filter) – AOBS (Acousto-Optical Beam Splitter) System. Scanning stage for motorized stage applications.
• Confocal Laser Scanning Microscope (CLSM) LEICA TCS SP2 fitted with seven excitation lines (457, 477, 488, 496, 514, 543, 633 nm). AOTF and AOBS.
• Widefield Multidimensional Microscopy System Leica AF6000 LX for live cell imaging (epifluorescence and transmitted light). It consists of an inverted Leica DMI6000B microscope and an incubation chamber equipped with CO2 and temperature controls. System equipped with two cameras: CCD Hamamatsu 9100-O2 and Orca-Flash 4.0 LT Digital CMOS camera, this one fitted with an image splitting optics (W-VIEW GEMINI), which provides one pair of dual wavelength (GFP/mCherry) images separated by a dichroic mirror onto a single camera. Scanning stage for motorized applications.
• The facility is equipped with a high capacity storage system for digital images, which is password-accessible from any other workstation connected to the CIB intranet
• The facility also hosts an independent image workstation equipped with different software, including Metamorph offline (Molecular Devices), for image analysis and data output.
Fluorochromes: Furthermore, there are various conjugated fluorescent antibodies available for the users at reduced price (100 μl aliquots).
Clients belonging to the Centro de Investigaciones Biológicas (CIB) have to book in the intranet calendar according to standards that are exposed on the reservations page.
External clients to CIB can contact by e-mail (firstname.lastname@example.org ) or phone ( 034 918 373 112 ext. 4401). The person from the facility who gets the message, will make booking at the intranet calendar. The Client interested on use any equipment of the facility must provide information to refill an Offer of Service that will be sent to him by the administration department of the Center
Contact with the Department by phone or e-mail for booking. Timetable: from 8 AM to 17 PM (Monday to Thursday) and from 8 AM to 15 PM (Friday).
TECHNICAL SERVICES CURRENTLY OFFERED.
The main services offered by the Microscopy Unit to the scientific community are:
- Adquisition images in different scan modes: XY, Xt, XYZ, XZY, XYZʎ, XYZt
- 3D Reconstruction
- FRAP analysis (Fluorescence recovery after photobleaching),
- FRET analysis (Förster resonance energy transfer) and another dynamic analysis.
- colocalization analysis
- Motorized applications: Mark & Find and Tile Scan
Multidimensional Microscopy fluorescence Advanced System for in vivo studies
- Multidimensional acquisition of images in X,Y,Z,t and ʎ. Equipped with a wide range of fluorescence filters spanning whole visible spectrum from UV to near IR. Being fully motorized in temporal acquisition.
- FRET (Förster resonance energy transfer) studies with CFP_YFP tandem.
- Calcium in vivo studies with FURA and other dynamic studies.
- Motorized applications: Mark & Find enables the user to define multi position experiments to observe a large number of cells within one experiment. Is particularly useful for studying dynamic processes in live cells. And the Tile Scan feature allows the acquisition of overview images of the specimen with multiple channels and in three dimensions.
- Protein colocalization in vivo with high spatial and temporal resolution. Hamamatsu camera Orca Flash 4.0 LT has an effective number of pixels of 2048 x2048 with a cell size of 6,5 µm x6,5 µm, and a minimum frame rate of 30 frames per sec (30 fps), and can even reach values above 1000 fps. In addition, it has attached an image splitting optics (W-VIEW GEMINI), which provides one pair of dual wavelength (GFP/mCherry) images separated by a dichroic mirror. Simultaneous image acquisition of dual wavelength images allows you protein colocalization studies, high speed ratio metric imaging and other multiple fluorescence applications. Acquisition software Metamorph (Molecular Dynamics)
Epifluorescence microscope equipment with CCD camera
Light transmitted and epifluorescence Axioplan Universal Microscope Zeiss with contrasting techniques of bright field and phase contrast. Fluorescence with filters for UV, green and red. Digital Camera Leica DFC 350 FX
Stereo Microscope Leica MZ16 FA
Objective 1× AN 0,14 /oculars 10×. Zoom capability (16:1):7.1× – 115×. Resolution 420Lp/mm. Field of view about 29,6mm – 1,8mm (according to zoom system). 8 Mpx Camera in color Leica DFC490. It allows the observation of samples and the capture of images both, with visible light (diascopy base with Rottermann contrast, bright field, dark field and cold light source), as with fluorescence light (UV, GFP, TXRed). Acquisition of images in different resolutions and formats.
The facility also hosts an independent image workstation equipped with different software, including Metamorph offline (Molecular Devices), for image analysis and data output.
The facility is equipped with a high capacity storage system for digital images, which is password-accessible from any other workstation connected to the CIB intranet
The services offered include the assessment to the users on all aspects related to the technique, even the experimental design and the data analysis.