NATIONAL CANCER INSTITUTE - CANCER.GOV

Contact Information

Primary Contact

Sandeep Pallikkuth
Core Head

Location

9000 Rockville Pike
Building 37, Room 2033B
Bethesda, MD 20892

Additional Contacts

Itoro Akpan
Biologist

Overview

LCMB Microscopy Core offers live cell imaging technologies as well as super-resolution, fluorescence lifetime and confocal imaging systems for immunofluorescence. Our confocal instruments are a Leica SP8 laser scanning confocal microscope and a Nikon spinning disk confocal microscope. We also house a Total Internal Reflection Fluorescence (TIRF) microscope with capability to perform super-resolved Single Molecule Localization Microscopy and incuabtion chamber equipped epi-fluorescence imaging systems for long-term live-cell imaging experiments.

Established Technologies
  • Live cell imaging, super-resolution imaging, confocal imaging, FLIM, FRET, single-molecule localization imaging/tracking.
  • Equipment includes Leica SP8 laser scanning confocal microscope with Falcon-FLIM and Lightning modules, a Nikon spinning disk confocal microscope with laser stimulation/FRAP module, and a Nikon epifluorescence/TIRF microscope with N-STORM and an Olympus long-term live-cell imaging epi-fluorescence microscope. The core also houses general-use bright-field/epi-fluorescence optical microscopes and a stereo microscope. We also have 3 imaging data processing work stations.
Developing Technologies
  • We are constantly developing analysis software for FLIM, SMLM and single-molecule tracking image data, customized for cancer research imaging experiments.

Major Instrumentation

  • Leica SP8 scanning confocal microscope, equipped with Falcon-FLIM module (for FLIM and FRET-FLIM experiments), and Leica Lightning module (for fast simultaneous multi-color super-resolution fluorescence imaging).
  • Nikon Eclipse Ti2 inverted microscope equipped with Yokogawa CSU-X1 spinning disk confocal system, 3-channel simultaneous confocal detection.
  • Nikon Eclipse TE2000 inverted microscope with total internal reflection fluorescence (TIRF), and super-resolution fluorescence detection (N-STORM) modules.
  • Olympus XI81 epifluorescence microscope with an enclosed incubator and a precise motor-controlled stage.
  • The Core also houses a Keyence all-in-one automated epifluorescence microscope, general use bright-field/epi-fluorescence microscopes with color camera detection, and a stereo microscope.
  • Three workstations for image processing with high-quality graphic cards, large RAM capacity, and high-speed solid-state drives for rapid processing of large 3D and 4D image data.

User Guidelines

The microscopes at the LCMB Microscopy Core are available for use to all members of NCI, free of cost. Facility staff provide training to all NCI users on the instruments at the LCMB Microscopy Core. Advanced NCI users, fully trained and cleared for independent use by facility staff, are free to use the microscopes by themselves after  scheduling time slots on the calendar. However, only LCMB members can use the instruments at times when the Facility staff are not present (i.e., evenings and weekends). Access and training are also provided to non-NCI users with a fee.

Publications

  • L. Balagopalan, T. Moreno, H. Qin, B. C. Angeles, T. Kondo, J. Yi, K. M. McIntire, N. Alvinez, S. Pallikkuth, M. E. Lee, H. Yamane, A. D. Tran, P. Youkharibache, R. E. Cachau, N. Taylor, L. E. Samelson, u201cGeneration of antitumor chimeric antigen receptors incorporating T cell signaling motifsu201d, Sci. Signal., 17 eadp8569 (2024).
  • K. Raychaudhuri, R. Rangu, A. Ma, N. Alvinez, A. D. Tran, S. Pallikkuth, K. M. McIntire, J. A. Garvey, J. Yi, L. E. Samelson, u201cCD28 shapes T cell receptor signaling by regulating ZAP70 activation and Lck dynamicsu201d, BioRxiv (2024).
  • E. M. Rosenberg Jr., X. Jian, O. Soubias, H. Y. Yoon, M. P. Yadav, S. Hammoudeh, S. Pallikkuth, I. Akpan, P. W. Chen, T. K. Maity, L. M. Jenkins, M. E. Yohe, R. A. Byrd, P. A. Randazzo, u201cThe small molecule inhibitor NAV-2729 has a complex target profile including multiple ADP-ribosylation factor regulatory proteinsu201d, J. Biol. Chem., 299(3), 102992 (2023).
  • Barr, V.A., Piao, J., Balagopalan, L., McIntire, K.M., Schoenberg, F.P., and Samelson, L.E. Heterogeneity of Signaling Complex Nanostructure in T Cells Activated via the T Cell Antigen Receptor. Microscopy and Microanalysis, 29, 1503, 2023. https://doi.org/10.1093/micmic/ozad072
  • Nunes-Santos, C.J., Kuehn, H., Boast, B., Hwang, S., Kuhns, D.B., Stoddard, J, Niemela, J.E., Fink, D.L., Pittaluga, S., Abu-Asab, M., Davies, J.S., Barr, V.A., Kawai, T., Delmonte, O.M., Bosticardo, M., Garofalo, M., Carneiro-Sampaio, M., Somech, R., Gharagozlou, M., Parvaneh, N., Samelson, L.E., Fleisher, T.A., Puel, A., Notarangelo, L.D., Boisson, B., Casanova, J.-L., Derfalvi, B., and Rosenzweig, S.D. Inherited ARPC5 deficiency is an actinopathy imparing cell motility and dissecting cytokine signaling. Nat.Commun., 2023, 14(1):3708. doi: 10.1038/s41467-023-39272-0, 2023.
  • Gasparski AN, Moissoglu K, Pallikkuth S, Meydan S, Guydosh NR, Mili S., mRNA location and translation rate determine protein targeting to dual destinations. Mol Cell. 2023 Aug 3;83(15):2726-2738.e9. doi: 10.1016/j.molcel.2023.06.036. Epub 2023 Jul 27. PMID: 37506697
  • Moissoglu K, Lockett SJ, Mili S., Visualizing and Quantifying mRNA Localization at the Invasive Front of 3D Cancer Spheroids, Methods Mol Biol. 2023;2608:263-280. doi: 10.1007/978-1-0716-2887-4_16. PMID: 36653713

Keywords

CO2FRAPFRETLeica SP8 laser scanning confocal microscopeNikon epifluorescence/TIRF microscopePALMTIRFTotal Internal Reflection FluorescenceconfocalImaging and Microscopyimmunofluorescencelocalization microscopymulti-labellingnci-coreoptical microscopyphotoactivated localization microscopyphotoactivationspinning disk confocal microscopetilingtime lapse with heattotal internal reflection fluorescenceFLIMsingle molecule localization microscopysuper-resolution fluorescence microscopy