Cancer Institute A national cancer institute
designated cancer center

Cell Sciences Imaging Facility (CSIF)

https://sharedfacilities.stanford.edu/service_center/show_external/1/CSIF

Facility Director: Jon Mulholland
Facility Advisor: Tim Stearns

Contact

Jon Mulholland
Director, Cell Sciences Imaging Facility
Beckman Center B050
Stanford, CA 94305
(650) 725-7532
E-mail:  jwm@stanford.edu

Overview

The Cell Sciences Imaging Facility (CSIF) provides access to and training in high-resolution, state-of-the-art fluorescence and electron microscopy services. The facility has three advanced imaging systems that provide confocal, deconvolution and 2-photon fluorescent light microscopy technologies. For electron microscopy, the facility has a full-service laboratory that offers sample preparation and training for both transmission and scanning electron microscopy technologies. Specific goals include acquisition of cutting-edge light and electron imaging instrumentation, training in the proper application and use of available equipment and technical assistance and consultation in the application of advanced immuno- and histo-chemical techniques. The CSIF is also dedicated to providing access to multi-dimensional (3D, 4D) imaging technology for volumetric analysis of cell and tissue architecture and continues to obtain and make available advanced 3D, 4D volume rendering, image-analysis software.

Services

Fluorescence Microscopy Services

The CSIF provides training and consultation in the use of confocal, deconvolution and 2-photon fluorescent light microscopy technologies. Major instrumentation available for use includes a Zeiss LSM 510 NLO 2-photon confocal microscope with a Non Descanned Detection system, a Leica SP2 AOBS confocal and an API Delta Vision wide-field deconvolution system. Confocal, 2-photon and deconvolution technologies allow optical sectioning while eliminating out-of-focus fluorescence. Additionally, time-lapse software allows 3D localization of labeled protein over time, thus providing 4D data sets. The confocal microscope is also equipped with a prism spectrometer detector which allows spectral detection and un-mixing of overlapping spectrum, e.g., eCFP, eGFP and eYFP expressed in the same cell. The CSIF’s LSM510 confocal system is equipped with a new high-sensitivity, non-descanned detection system, which allows researchers to acquire images from samples that are less fluorescent or from deeper in tissue. Thus, 2-photon excitation, together with non-descanned detection is ideal for live cell imaging of thick tissue samples.,

Electron Microscopy Services

The CSIF offers a full-service laboratory with sample preparation and training for both transmission and scanning electron microscopy technologies. Sample preparation services include fixation, microtome and staining procedures as needed. The facility houses a transmission electron microscope that can produce a resolution of 2 angstroms (10-10). It is fitted with a high-contrast, biological objective lens, making it ideal for imaging thin, immuno-localized samples used for the unambiguous determination of a protein's subcellular location. The facility’s scanning electron microscope is used for study of specimen topology.

Data Management

The CSIF provides all data management needs of all users. This includes server storage (1.25 terabytes) and tape data archiving. Each user of the CSIF has a password-protected computer account and the user is able to access to data via secure FTP. Data are saved directly to the server as acquired and backed up to tape nightly; it is the CSIF policy to hold backed-up data (i.e., recoverable from tape) for one year.

Operation

The CSIF is located in the basement of Beckman Center. Trained and approved users of the facility have access to the microscope during normal business hours (8 am to 5 pm, Monday through Friday); after-hours access, via an access code on the CSIF facility doors, is allowed after at least two independent sessions and with the approval of the CSIF director.

Procedures

Investigators contact the CSIF Director to discuss experimental needs and their microscopy background. Researchers then complete an online application containing information the CSIF requires for account setup. Once this form has been received, an appointment for training or experimental procedure is made with the CSIF staff. Training typically requires two three-hour training sessions, during which time trainees are taught the proper, safe use of the microscope and how to optimize imaging for their specific experimental goals.

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