Confocal microscopy

Latest news

* LSM800 out of service. (18/11/2020)

Simultaneous acquisition on Spinning Disk equipment is NOT available. Sequential acquisition works correctly. (11/11/2020)

* Installed new PCO edge 4.2 bi camera in FRET equipment. (09/09/2020)

Scientific supervisor: Francisco Javier Díez-Guerra
Technical director: Ángeles Muñoz

  confocal-cbm (at) listas.csic.es
  Third floor - 310

The optical and confocal microscopy facility (SMOC) is the responsible of maintaining and managing the advanced-optical-microscopy equipment in the CBMSO. It also offers support, advice and training in optical microscopy techniques and image analysis, it distributes reagents and other materials related to microscopy, and is responsible for finding resources to acquire new equipment that suits the requests of CBM researchers.

Our Facility, as a central scientific service at the Centro de Biología Molecular "Severo Ochoa", does not carry out any scientific research work but offers:

  • 7 confocal microscopy systems and 6 wide-field microscopes, with different levels of automation, allowing studies on live and fixed samples. In addition, 2 workstations for image analysis, a vibratome and a stereomicroscope.
  • An online platform to schedule and manage the equipment and invoicing.
  • A stock of reagents and materials useful for microscopy.
  • A quality management system ISO 9001-2015 certified by AENOR, since March 2009.

SMOC:

PUBLICATIONS:

  • "Setting up and running an Advanced Light Microscopy and Imaging Facility". Carlos Sánchez, Mª Ángeles Muñoz, Maite Villalba, Verónica Labrador and F. Javier Díez-Guerra 2010. Current Protocols in Cytometry. 57:12.22.1-12.22.21.
  • "Técnicas avanzadas de microscopía óptica para el estudio de la fisiopatología celular". Francisco Javier Díez Guerra y Carlos Sánchez Martín. Capítulo 5, Sección II (Papel de la Biología Molecular y Celular en el estudio de las Enfermedades Renales). Nefrología Clínica. (4ª Edición. Noviembre 2013). Autor: Manuel Arias Rodríguez. Editor: L. Hernando Avendaño. Editorial Médica Panamericana
Image

* For external calls please dial 34 91196 followed by the extension number
Last name Name Laboratory Ext.* Schedule e-mail Professional category
Calvo Cazalilla Elena 310 4643 9:00 - 16:30 elena.calvo@cbm.csic.es Tco. Sup.Investig. y Laboratorio, GP3
Gallego García Carlos 310 4643 8:00 - 15:30 cgallego(at)cbm.csic.es Tit.Sup.Activ.Técn.y Profes. GP1
Muñoz Alcalá Mª Angeles 310 4643/4613 9:00 - 16:30 mamunoz(at)cbm.csic.es E.Ayudantes De Invest. De Los Oo.Publicos De Investigacion
Sánchez Jiménez Carmen 310 4643 9:30-17:00 csjimenez(at)cbm.csic.es Titulado Sup.de Actividades Técn. y Profes. GP1
Vega Sabugo Francisco José 310 4643 9:30 - 17:00 fjvega(at)cbm.csic.es Titulado Sup. Actividades Tecn. y Prof.GP1
Villalba Villacorta María Teresa 310 4643 8:00 - 15:30 tvillalba(at)cbm.csic.es Ayudante Investigación
Widefield Microscopes
...
sCMOS-monochrome Camera

Location: 3rd Floor (Lab. 310)

In vivo system in the inverted microscope Axiovert200 (Zeiss) coupled to a monochrome sCMOS camera and ultrafast filter switching

Booking

Technical Specifications

USER GUIDE (Disponible en "Documents" del sistema de Reservas)

Image Calibration: Pixel/µm size relationship

Deconvolution: Necessary values for a correct acquisition

...
F.R.E.T
 

Location: 3rd Floor (Lab. 310)

In vivo system in the inverted microscope Axiovert200 (Zeiss) coupled to a monochrome sCMOS camera and ultrafast filter switching

Booking

Technical Specifications

USER GUIDE (Available through "Documents" at the Booking System)

Image Calibration: Pixel/µm size relationship

Deconvolution: Necessary values for a correct acquisition

...
High-speed in vivo system
 

Location: 4th Floor (Lab. 423.a)

In vivo system in the inverted microscope AF6000 LX (Leica) coupled to a monochrome EMCCD camera and ultrafast filter switching for calcium experiments.

Booking

Technical Specifications

USER GUIDE (Available under "Documents" at the Booking System)

Image Calibration: Pixel/µm size relationship

Leica System File Processing Program

Deconvolution: Neccessary values for a correct acquisition

Calcium Measurement

...
CMOS-color Camera
 

Location: 3rd Floor (Lab. 335)

Upright Axioskop2 plus microscope (Zeiss) coupled to a color CMOS camera (DMC6200 Leica)

Booking

Technical Specifications

USER GUIDE (Available under "Documents" at the Booking System)

Image Calibration: Pixel/µm size relationship

...
CoolSNAPfx – monochrome Camera

Location: 3rd Floor (Lab. 335)

Uprigth Axio Imager M1 Microscope (Zeiss) coupled to a CCD monochrome camera (Coolsnap FX)

Booking

Technical Specifications

USER GUIDE (Available under "Documents" at the Booking System)

Image Calibration: Pixel/µm size relationship

 
Image
Available reagents at the Confocal Microscopy Facility
General recommendations - Tips
Fluorophores and fluorescent proteins
  • Check the excitation and emission spectra of the florophores and fluorescent proteins in order to optimize the method used for imaging.
  • When a fluorescent protein (FP) is used for the first time check the emission spectrum, by spectral detection, to ensure the real identity of the protein.
  • The laser line 405 nm may photoconvert your FP (change the emission profile). Check it.

Servicio de Microscopía Optica y Confocal (Lab.310)

Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM)

C/Nicolás Cabrera,1

Universidad Autónoma de Madrid. Cantoblanco.

28049. Madrid

Teléfonos:

Despacho: 91 196 4613

Laboratorio 310: 91 196 4643

Laboratorio 336: 91 196 4660

Fax. 91 196 4420

E-mail: confocal-cbm@listas.csic.es

SMOC en 3 plantab 1

Transporte en la Comunidad de Madrid

Transporte en la U. Autónoma de Madrid

RENFE

Líneas C7, C8 y C10 de Cercanías: Parla-Atocha-Chamartín-Cantoblanco-Alcobendas/S.S. de los Reyes o Colmenar Viejo.

Estación de Cantoblanco.

AUTOBUSES INTERURBANOS

Líneas 714, 827, 827B y 828

METRO DE MADRID Y LÍNEAS DE AUTOBUSES EMT

Protocols
Autofluorescence
Methods for removing autofluorescence
  • Sodium Borohydride (NaBH4)
    • Treat the fixed coverslips with NaBH4 (1 mg/ml in PBS, pH 8.0) for 10 minutes at room temperature (RT)
    • Wash with PBS and continue with the immunofluorescence protocol. This solution must be recently prepared, observing bubble formation

  • Toluidine blue (‎C15H16N3ClS)
    In cases that FITC is the detection method and mainly for tissues. This will decrease autofluorescence mainly due to chlorophyll and cell walls (elastin/collagen):
  • Ammonium chloride (NH4Cl)
    • Treat the fixed coverslips with NH4Cl (50mM diluted in PBS, pH 8.0) for 10 minutes at RT
    • Wash with PBS and continue with the immunofluorescence protocol

  • Sudan Black (C29H24N6)
    • Treat the coverslips after secondary antibodies incubation with 0.3% Sudan Black (w/v) in 70% EtOH (v/v) for 10 minutes
    • Wash extensively with PBS and proceed mounting the samples

  • Trypan Blue (C34H28N6O14S4)
    • Treat the coverslips after secondary antibodies incubation with 250µg/ml Trypan Blue in PBS pH 4.4, for 1 minute
    • Wash extensively with PBS and proceed mounting the samples
Rates
Image
Image

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