Overview of the UW-Madison Genetics Confocal Facility

All users receive training on how to use the Zeiss 510 Confocal tailored to their prior knowledge before being allowed to use to the system. Training is directed towards producing independent users. Sample preparation is solely the users responsibility. Advice is available but the facility personnel do not perform experiments, gather data for users or are responsible for scientific outcome. Use of the confocal scanner is charged on a fee per unit time of use basis.

Training in the independent use and proper care of the confocal microscope and the associated computer equipment and software, some advice in experimental design and assistance in optimal use of the equipment and image processing and handling are provided. Once trained and checked out, users are allowed independent access to the equipment. Training, which involves major contact time, is done on specific days determined by the main trainer of your particular lab.

The facility is under the management of Dr. Barry Ganetzky, with Dr. Richard Daniels serving as a trainer and management personnel.

Barry Ganetzky ganetzky@wisc.edu Office Phone: 263-2404
Richard Daniels rwdaniels@wisc.edu Lab Phone: 262-3896

Things to do to gain access to the Genetics Confocal: Get at least a 2-day training session from the confocal trainer for your lab Sign up for confocal calendar access once trained Request confocal room access by emailing Ganetzky lab for approval Make sure you use the sign in sheet in the room every time you use the system.

Calendar Sign Up
A web-based booking calendar system is provided for the facility, and is available from any location to all authorized users via shared WiscCal.  Please email Richard Daniels (rwdaniels@wisc.edu) for access and instructions for adding the calendar and booking time.

Confocal Room Access
Once you have been trained for use of the confocal, please tell Richard Daniels or Barry Ganetzky and they will get your campus ID approved for access to the confocal room. Marj Haanstad (haanstad@wisc.ed) will grant access after she receives approval from either Richard or Barry.

Transferring files
FTP transfer is available within the department. Please obtain a login and password from your lab to access the AKKA server.http://akka.genetics.wisc.edu Transfer your files to a computer in your lab asap.WARNING: Any files left on the confocal computer for more than a month will be dumped.

Laser Excitation Spectrum
Argon: 458nm, 477nm, 488nm, 514nm
Diode: 405nm (DAPI)
HeNe: 543nm, 643nm
DPS 561: 561nm

Common Uses for our Lasers
405-UV or DAPI
458-CFP
477-FITC
488-GFP, ALEXA 488
514-YFP
561-Rhodamine, ALEXA 568, CY3
633-ALEXA 647, CY5

Table of Fluorochromes:http://pingu.salk.edu/flow/fluo.html
Molecular Probes Spectra Viewer:http://probes.invitrogen.com/resources/spectraviewer/

Objectives
4X, 10X, 20X, 40X Oil, 63X Oil, 100X Oil

Mercury lamp
DAPI, FITC and Rhodamine dichroic filters are available for sample evaluation and subject location.

DIC
DIC is available. Transmitted images can be obtained along with your confocal images.

Usage charges
Only Laboratory of Genetics labs and Genetics Trainer labs have access to this confocal.

Use of the general purpose PC and Confocal PC for image analysis processing and preparation (no confocal laser scanning) are provided without charge to authorized users.

All users/trainees are charged at the same rate regardless of Laboratory or Department.

Computers & Usage
The Windows computers in the facility maintain extensive logs of usage of the confocal equipment and ancillary computers used for image processing. The information from these logs is passed on to the Genetics Department business office, where personnel prepare bills for the confocal scanner time accrued by users. PIs will be billed via the usual UW-Madison billing channels.

Notes

Initial capital funding for the Zeiss 510 Confocal system was provided by a NSF Grant in 2007, by Barry Ganetzky, PI; Ahna Skop, Francisco Pelegri, Xin Sun, Aki Ikeda, Jerry Yin, co-applicants. Additional funding was provided by the Genetics Department.