Office for Research and Graduate Studies (ORAGS)

Confocal Microscopy Core Facility

Scientific Objectives and Instrumentation:  The aims of the Confocal Core are to offer state-of-the-art microscopy and imaging capabilities to campus researchers, and to help improve the microscopy-related skills of these scientists. 

Since the core was founded 10 years ago, 230 researchers have participated in the individual 4-hour training session required to operate the Zeiss LSM410 confocal microscope system, which has been the core’s primary instrument. The user group represents 28 campus departments, and includes scientists from the Schools of Medicine, Dentistry, Nursing and Pharmacy, as well as other units on campus, including the MBC, IHV, and VA.   The large majority, however, has been faculty, students and staff in the School of Medicine.  The facility has been extremely active, with nearly 10,000 hours of use since its inception.

The new Zeiss LSM 510 META is a hyperspectral confocal microscope.  It can separate fluorophores with overlapping emission spectra based on their spectral signatures, by measuring up to 32 emission wavelength bands.  It also has a Ti:S laser for multiphoton applications in addition to 453, 488, 514, 543, and 633 excitation laser lines. 

Personnel:  The Facility for Confocal Microscopy is managed by it’s Director (P. Luther, Assistant Professor, Dept. of Physiology), who is responsible for maintaining the instrument, training new users, consulting with researchers regarding the technical aspects of sample preparation, advanced confocal techniques, data analysis, etc. 

Training:  Individuals who require the instrument go through a basic training module, supplemented by various advanced modules for special techniques.  These sessions, which are performed one-on-one, are arranged by appointment.  The basic module is an 4-6 hour session that covers a) start-up procedures (5 min), description of optical path (60 min), which includes function of components, discussion of detector pinhole operation and parameters (size, alignment), concepts of numerical aperture and spatial sampling, signal optimization and spectral overlap problems; c) software and imaging (90 min), including image optimization, noise reduction, factors affecting resolution and choice of laser power, and image analysis; d) a free-time session (30 min), in which users operate the instrument independently to experiment with the various controls and prove to themselves that they are capable of operating the instrument without supervision. e) pinhole alignment (20 min), and introduction to 3D reconstruction.  The session also addresses specimen preparation, and users are expected to supply samples so their suitability for confocal techniques can be assessed.  The session is high-paced, but we have developed a series of training aids to ensure that the concepts are understood.  According to the requirements of the investigator, additional training modules are available for z-series and 3D reconstruction, multicolor confocal microscopy, laser scanning DIC and interference reflection imaging, microscopy of living cells using the water immersion objective, high speed and xt scanning, and image analysis.  We have created a 30-page “Users Guide”, which provides specific instructions for operating the instrument (see website).  This document is supplemented by a series of handouts that cover various advanced aspects of confocal microscopy.

Contact Information, location, hours of operation:

Paul Luther, Ph.D.      6-4166
pluther@umaryland.edu

Zeiss 410        Howard Hall 433          Available 24/7

Zeiss 510        Howard Hall 525          Available 24/7

Updated Pricing:

Zeiss 410:       $500.00 training (includes first five hours of scope time and must be used within 30 days of training), $35.00 per hour.

Zeiss 510:       $500.00 Training, $40.00 per hour (New service, price may be subject to change)

Oversight Committee Members:

Bloch, Robert J. (Physiology)
Lederer, W. Jonathan (MBC)
Pallone, Thomas L. (Nephrology)
Rizzo, Mark A. (Physiology)
Wade, James T. (Physiology)

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