Creating Biomedical Technologies to Improve Health

LABS AT NIBIB

Interferometric Photoactivated Localization Microscope (IPALM)

Capable of unsurpassed optical resolution, this system is a clone of the original prototype developed at the Janelia Research Campus, but offers 3 color capability. IPALM combines photoactivated localization microscopy with single-photon, simultaneous multiphase interferometry providing <20 nm resolution in all three dimensions. This microscope is currently located in the Taraska Lab and will move to AIM once the facility is operational.

Donating lab: Justin Taraska, NHLBI

Spatial resolution: 20nm in X, Y. 10nm in Z.

Temporal resolution: Fixed samples only.

Excitation wavelengths: 488nm, 647nm, 750nm.

Detection channels: Single channel (3 cameras used for interferometric localization of single emitters). Multiple emission bands are acquired sequentially.

Sample specifications: Fixed samples sandwiched between two coverslips <10 microns apart. The sample is placed between two objectives in a 4Pi geometry and images can be acquired up to 750nm from the coverslip surface. This is ideally suited for high resolution imaging of membrane or peri-membrane structures.

Strengths:

  • Highest resolution optical microscope currently available.
  • Provides unparalleled axial resolution.

Limitations:

  • Difficult sample preparation protocol.
  • Sample thickness is restricted and only sub-volumes of mammalian cells can be imaged.

Reference:
Shtengel et al. PNAS 106(9): 3125-3130, Mar 2009.