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Leica SP5 Scanning Confocal

Location: U4204 Learned Lab

Reservations required on VU iLab website

This confocal is reserved for Primary and Joint CDB Faculty and their personnel.

Consultation with the Equipment Manager is required before using this microscope.


Want a Leica .lif Viewer?

Please visit Leica. Then, go to the bottom of the page. You will see an option to download “LAS AF LITE” above the Apple icon.


The Scope

The Leica SP5 confocal is a DMI 6000 model Inverted microscope with a Super Z Galvo stage for fast Z sectioning and prisms for DIC imaging. Microscopists utilize the scanning stage for setting up multiple z-stacks with the Mark-n-Find option and for stitching numerous fields into one large image. The galvo is detachable to fit a humidified incubation chamber for time-lapse or cross-section live cell imaging. A heating collar for the 100x 1.47 NA objective provides additional improvement in the optics by mitigating thermal gradients.

The Leica SP5 is supported by an Argon laser for excitation at 458, 476, 488, 496, and 514 nm; a DPSS for 561 nm and a HeNe for 633 nm. Users benefit from simultaneously collecting 3 color images (+ DIC) with PMTs or up to 6 colors with sequential acquisition function. Addition of a GaAsP hybrid detection system (HyD) significantly improves image quality and detection efficiency. This HyD technology delivers super-sensitivity which improves cell viability and offers low dark noise while providing a larger dynamic range. FRAP and FRET Wizards are included in the software for increased functionality.

Leica Reservation Policy

The Leica has been a heavy workhorse for confocal microscopy in our department. This is a result of an increasing number of CDB personnel who are now being trained to use it. In an attempt to make the reservation/billing procedures fair to all and to maximize the number of reservation slots available, the Equipment Resource Committee is enacting the following guidelines as of February 25, 2013.

  • All bookings will be charged in full unless canceled 48 hours in advance.
  • If you are unable to make your reservation within the 48 hours and if you are able to locate someone to fill your spot, then the reservation transfers to them and you will not be billed. However, the person taking over the reservation must email the Equipment Manager confirming the arrangement.
  • Bookings can only be made two weeks in advance.
  • Currently, there is no maximum amount of time the instrument can be booked. However, booking times could be limited in the future if the needs of the business change.

Additional Information for Download

New users are strongly encouraged to read the TCS SP5 user manual chapter titled “Introduction to confocal work” which discusses many issues of confocal microscopy in a clear and direct manner. Users may also benefit from reading “What is Köhler illumination?”, especially if the term is not familiar. There are additional .pdfs below which are good references for using the scope and loading your files into ImageJ.

Leica TCS SP5 User Manual.pdf

Leica Quickstart Guide.pdf

leica confocal microscope.pdf

Loading LIF files into ImageJ.pdf

Leica Reservation Guidelines.pdf