Instrumentation - Nikon AXr-Multiphoton Intravital Microscope

Specifications

Laser’s configurations:

  • Single-photon:
    • 405 nm
    • 488 nm
    • 561 nm
    • 637 nm
  • Multiphoton/two-photon:
    • Fixed wavelength 1045 nm
    • Tunable wavelength 820 – 1300 nm

Objectives available

CFI75 Apochromat 20x Water Immersion Objective (MRD77226) with Correction Collar. N.A.: 1, Working distance: 2.8mm, Cover glass thickness: 0–0.17.

CFI90 20XC Glycerin/Glycerol Immersion Objective (MRD71220) with Correction Collar for cleared tissue. N.A.: 1, Working distance: 8.2mm, Cover glass thickness: 0, Correction: 1.44 - 1.50.

Miscellaneous additions

Heated temperature-controlled black enclosure for reducing light contamination & maintaining animal temperature.

Fast-speed Resonant scanner with 4k imaging.

High-resolution Galvo scanner with 8k imaging.

This microscope is also suitable for imaging cleared tissue using either the water immersion or glycerol immersion objective. Please contact the core director to discuss imaging cleared tissue.

Anticipated Uses & Detailed Information:

This microscope is located within the animal facility in the Georgia Cancer Center (CN) Building, CN-5113. The use of this system is for fluorescence-based live animal (intravital) imaging of mice. As there is a large space on the stage that can accommodate considerable depth, other animals might also be feasible for imaging studies but consultation with the core director would be required for prior approval before such use.

The system has an integrated laser line system with both traditional single-photon confocal capabilities as well as a new two-laser multiphoton laser set up. The dual multiphoton has a fixed laser firing at 1045 nm and a tunable laser ranging from 820 – 1300 nm. The single-photon and multiphoton integrated system allows wide imaging usage capabilities depending on the research question to be answered.

The multiphoton is equipped with traditional Galvano-controlled sensors for high content imaging (8k resolution) or high-speed resonant scanner for fast imaging (2k resolution). This microscope system is from Nikon and uses a different software platform known as Nikon Elements.

For live animal imaging, users need to be trained on both microscope usage as well as animal surgery for self-use. Typically, there will be a total of three training sessions - two sessions for microscope use and one animal surgery demonstration (for typical organ/tissue imaging). After the animal surgery demonstration, the user is expected to practice animal surgery on their own before microscope use.