Ammasi Periasamy

Professor of Research, Biology

Primary Location

PO Box 400328

Primary Phone Number

434.243.7602

[email protected]

Website

http://www.cci.virginia.edu/

Bio & Overview

I designed and developed different light microscopy imaging systems to investigate cellular signaling such as calcium, pH, protein-protein interactions and also diagnostic system in clinical imaging. The system includes: Digitized video microscopy, digital deconvolution, confocal, multi-photon excitation, fluorescence lifetime (FLIM), time-resolved, anisotropy and fluorescence resonance energy transfer (FRET) microscopy. My current research area of interest is energy-based FLIM-FRET Microscopy and Spectroscopy in the determination of when and where specific proteins associate with one-another in living cells and tissues. Through the use of conventional fluorescence microscopy, proteins labeled with different fluorophores can be localized within fixed or living cell preparations. The fluorophores absorb light at one wavelength and emit light at another, longer wavelength. By using appropriate filters, it is possible to detect several different labeled proteins in the same preparation. However, the optical resolution of the light microscope limits determination of protein proximities to 0.2 micrometers. Resolving the relative proximities of proteins that exceeds the optical limit of the microscope is necessary to reveal the physical interactions between protein partners. This degree of spatial resolution can only be achieved in energy-based imaging using the technique of fluorescence resonance energy transfer (FRET) and fluorescence lifetime imaging (FLIM) microscopy. FRET is a process by which radiation-less transfer of energy occurs from a fluorophore in the excited state to an acceptor molecule in close proximity. The range over which resonance energy transfer can occur is limited to ~ 0.01 micrometers and the efficiency of energy transfer is extraordinarily sensitive to the distance between fluorophores. The measurement of FRET in the microscope provides a non-invasive approach to visualize the spatio-temporal dynamics of the interactions between protein partners in the living cell. The fluorescence lifetime is defined as the average time that a molecule remains in an excited state prior to returning to the ground state. Many currently available fluorescence microscopic techniques, such as confocal or multi-photon excitation, cannot provide detailed information about the organization and dynamics of complex cellular structures. In contrast, fluorescence lifetime imaging (FLIM) microscopy allows the measurement of dynamic events at very high temporal resolution and can monitor interactions between cellular components with very high spatial resolution as well. An important advantage of FLIM is that the absolute values of lifetimes are independent of the probe concentration, photobleaching, light scattering and the amount of excitation intensity. FLIM thus offers many opportunities for studying dynamic event of living cells. A major obstacle to implementation of energy-based spectroscopy in living cells has been the lack of suitable methods for specifically labeling intracellular proteins with the appropriate fluorophores. The cloning of the jellyfish green fluorescent protein (GFP) and its expression in a wide variety of cell-types has proven this fluorescent protein to be a versatile marker for both gene expression and protein localization in living cells. When illuminated by blue light, the jellyfish GFP yields a bright green fluorescence that does not require any cofactors, substrates, or additional gene products. GFP retains its fluorescent properties when fused to other proteins, allowing energy-based microscopy to be used to visualize dynamic changes in protein localization in intact cells.

Academic Information

Department: Biology

Academic Title: Professor of Research

Other Appointments

Professor of Research, Biomedical Engineering

Other Affiliations

Professor of Research, BIMS

Research Interests

Imaging of Proteins in Living (or fixed) Cells and Tissues Using Light Microscopy Systems

Research Disciplines

Biomedical Engineering
Biophysics

Selected Publications

2022

Alam, S. R., Wallrabe, H., Christopher, K. G., Siller, K. H., & Periasamy, A. (2022). Characterization of mitochondrial dysfunction due to laser damage by 2-photon FLIM microscopy. SCIENTIFIC REPORTS, 12(1). doi:10.1038/s41598-022-15639-z

Norambuena, A., Sun, X., Wallrabe, H., Cao, R., Sun, N., Pardo, E., . . . Bloom, G. S. (2022). SOD1 mediates lysosome-to-mitochondria communication and its dysregulation by amyloid-β oligomers. NEUROBIOLOGY OF DISEASE, 169. doi:10.1016/j.nbd.2022.105737

2019

Zaman, K., Knight, J., Hussain, F., Cao, R., Estabrooks, S. K., Altawallbeh, G., . . . Gaston, B. (2019). S-Nitrosylation of CHIP Enhances F508Del-CFTR Maturation. AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY, 61(6), 765-775. doi:10.1165/rcmb.2018-0314OC

Marozkina, N., Bosch, J., Cotton, C., Smith, L., Seekler, J., Zaman, K., . . . Gaston, B. (2019). Cyclic compression increases F508 Del CFTR expression in ciliated human airway epithelium. AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 317(2), L247-L258. doi:10.1152/ajplung.00020.2019

Dai, J., Xing, Y., Xiao, L., Li, J., Cao, R., He, Y., . . . Li, X. (2019). Microfluidic Disc-on-a-Chip Device for Mouse Intervertebral Disc-Pitching a Next-Generation Research Platform To Study Disc Degeneration. ACS BIOMATERIALS SCIENCE & ENGINEERING, 5(4), 2041-2051. doi:10.1021/acsbiomaterials.8b01522

Periasamy, A. (2019). Special Issue on Metabolism Cytometry A. CYTOMETRY PART A, 95A(1), 10-12. doi:10.1002/cyto.a.23712

Cao, R., Wallrabe, H., Siller, K., Alam, S. R., & Periasamy, A. (2019). Single-cell redox states analyzed by fluorescence lifetime metrics and tryptophan FRET interaction with NAD(P)H. CYTOMETRY PART A, 95A(1), 110-121. doi:10.1002/cyto.a.23711

2018

Norambuena, A., Wallrabe, H., Cao, R., Wang, D. B., Silva, A., Svindrych, Z., . . . Bloom, G. S. (2018). A novel lysosome-to-mitochondria signaling pathway disrupted by amyloid-β oligomers. EMBO JOURNAL, 37(22). doi:10.15252/embj.2018100241

Wallrabe, H., Svindrych, Z., Alam, S. R., Siller, K. H., Wang, T., Kashatus, D., . . . Periasamy, A. (2018). Segmented cell analyses to measure redox states of autofluorescent NAD(P)H, FAD & Trp in cancer cells by FLIM. SCIENTIFIC REPORTS, 8. doi:10.1038/s41598-017-18634-x

2017

Sonavane, P. R., Wang, C., Dzamba, B., Weber, G. F., Periasamy, A., & DeSimone, D. W. (2017). Mechanical and signaling roles for keratin intermediate filaments in the assembly and morphogenesis of Xenopus mesendoderm tissue at gastrulation. DEVELOPMENT, 144(23), 4363-4376. doi:10.1242/dev.155200

Alam, S. R., Wallrabe, H., Svindrych, Z., Chaudhary, A. K., Christopher, K. G., Chandra, D., & Periasamy, A. (2017). Investigation of Mitochondrial Metabolic Response to Doxorubicin in Prostate Cancer Cells: An NADH, FAD and Tryptophan FLIM Assay. SCIENTIFIC REPORTS, 7. doi:10.1038/s41598-017-10856-3

Hsu, K., Lee, T. -Y., Periasamy, A., Kao, F. -J., Li, L. -T., Lin, C. -Y., . . . Lin, M. (2017). Adaptable interaction between aquaporin-1 and band 3 reveals a potential role of water channel in blood CO₂ transport. FASEB JOURNAL, 31(10), 4256-4264. doi:10.1096/fj.201601282R

Sun, B. L., Palmer, L., Alam, S. R., Adekoya, I., Brown-Steinke, K., Periasamy, A., & Mutus, B. (2017). O-Aminobenzoyl-S-nitrosoglutathione: A fluorogenic, cell permeable, pseudo-substrate for S-nitrosoglutathione reductase. FREE RADICAL BIOLOGY AND MEDICINE, 108, 445-451. doi:10.1016/j.freeradbiomed.2017.04.008

2016

Periasamy, A., So, P. T. C., & Konig, K. (2016). Introduction. MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XVI, 9712. doi:10.1117/12.2239754

2015

Zaman, K., Sawczak, V., Zaidi, A., Butler, M., Bennett, D., Getsy, P., . . . Gaston, B. (2016). Augmentation of CFTR maturation by S-nitrosoglutathione reductase. AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 310(3), L263-L270. doi:10.1152/ajplung.00269.2014

Wallrabe, H., Sun, Y., Fang, X., Periasamy, A., & Bloom, G. S. (2015). Three-color confocal Forster (or fluorescence) resonance energy transfer microscopy: Quantitative analysis of protein interactions in the nucleation of actin filaments in live cells. CYTOMETRY PART A, 87A(6), 580-588. doi:10.1002/cyto.a.22651

Burdikova, Z., Svindrych, Z., Hickey, C., Wilkinson, M. G., Auty, M. A. E., Samek, O., . . . Sheehan, J. J. (2015). Application of advanced light microscopic techniques to gain deeper insights into cheese matrix physico-chemistry (vol 95, pg 687, 2015). DAIRY SCIENCE & TECHNOLOGY, 95(6), 921. doi:10.1007/s13594-015-0262-1

Burdikova, Z., Svindrych, Z., Pala, J., Hickey, C. D., Wilkinson, M. G., Panek, J., . . . Sheehan, J. J. (2015). Measurement of pH micro-heterogeneity in natural cheese matrices by fluorescence lifetime imaging. FRONTIERS IN MICROBIOLOGY, 6. doi:10.3389/fmicb.2015.00183

2014

Sun, Y., & Periasamy, A. (2015). Localizing Protein-Protein Interactions in Living Cells Using Fluorescence Lifetime Imaging Microscopy. ADVANCED FLUORESCENCE MICROSCOPY: METHODS AND PROTOCOLS, 1251, 83-107. doi:10.1007/978-1-4939-2080-8_6

Periasamy, A. (2014). Advanced Light Microscopy. METHODS, 66(2), 121-123. doi:10.1016/j.ymeth.2014.03.011

Rehman, S., Gladman, J. T., Periasamy, A., Sun, Y., & Mahadevan, M. S. (2014). Development of an AP-FRET Based Analysis for Characterizing RNA-Protein Interactions in Myotonic Dystrophy (DM1). PLOS ONE, 9(4). doi:10.1371/journal.pone.0095957

2013

Wallrabe, H., Cai, Y., Sun, Y., Periasamy, A., Luzes, R., Fang, X., . . . Bloom, G. S. (2013). IQGAP1 Interactome Analysis by In Vitro Reconstitution and Live Cell 3-Color FRET Microscopy. CYTOSKELETON, 70(12), 819-836. doi:10.1002/cm.21146

Sun, Y., Rombola, C., Jyothikumar, V., & Periasamy, A. (2013). Forster resonance energy transfer microscopy and spectroscopy for localizing protein-protein interactions in living cells. CYTOMETRY PART A, 83(9), 780-793. doi:10.1002/cyto.a.22321

Jyothikumar, V., Sun, Y., & Periasamy, A. (2013). Investigation of tryptophan-NADH interactions in live human cells using three-photon fluorescence lifetime imaging and Forster resonance energy transfer microscopy. JOURNAL OF BIOMEDICAL OPTICS, 18(6). doi:10.1117/1.JBO.18.6.060501

Abe, K., Zhao, L., Periasamy, A., Intes, X., & Barroso, M. (2013). Non-Invasive In Vivo Imaging of Near Infrared-labeled Transferrin in Breast Cancer Cells and Tumors Using Fluorescence Lifetime FRET. PLOS ONE, 8(11). doi:10.1371/journal.pone.0080269

2012

Wallrabe, H., Sun, Y., Fang, X., Periasamy, A., & Bloom, G. (2012). Three-Color FRET expands the ability to quantify the interactions of several proteins involved in actin filament nucleation. MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XII, 8226. doi:10.1117/12.906432

Periasamy, A., Vogel, S. S., & Clegg, R. M. (2012). FRET at 65: A Celebration of Forster. JOURNAL OF BIOMEDICAL OPTICS, 17(1). doi:10.1117/1.JBO.17.1.011001

Sun, Y., Hays, N. M., Periasamy, A., Davidson, M. W., & Day, R. N. (2012). MONITORING PROTEIN INTERACTIONS IN LIVING CELLS WITH FLUORESCENCE LIFETIME IMAGING MICROSCOPY. IMAGING AND SPECTROSCOPIC ANALYSIS OF LIVING CELLS: OPTICAL AND SPECTROSCOPIC TECHNIQUES, 504, 371-391. doi:10.1016/B978-0-12-391857-4.00019-7

2011

Williams, M., Burdsal, C., Periasamy, A., Lewandoski, M., & Sutherland, A. (2012). Mouse primitive streak forms in situ by initiation of epithelial to mesenchymal transition without migration of a cell population. DEVELOPMENTAL DYNAMICS, 241(2), 270-283. doi:10.1002/dvdy.23711

Sun, Y., Day, R. N., & Periasamy, A. (2011). Investigating protein-protein interactions in living cells using fluorescence lifetime imaging microscopy. NATURE PROTOCOLS, 6(9), 1324-1340. doi:10.1038/nprot.2011.364

2010

Sun, Y., Wallrabe, H., Seo, S. -A., & Periasamy, A. (2011). FRET Microscopy in 2010: The Legacy of Theodor Forster on the 100th Anniversary of his Birth. CHEMPHYSCHEM, 12(3), 462-474. doi:10.1002/cphc.201000664

Sun, Y., Wallrabe, H., Booker, C. F., Day, R. N., & Periasamy, A. (2010). Three-Color Spectral FRET Microscopy Localizes Three Interacting Proteins in Living Cells. BIOPHYSICAL JOURNAL, 99(4), 1274-1283. doi:10.1016/j.bpj.2010.06.004

Sun, Y., & Periasamy, A. (2010). Additional correction for energy transfer efficiency calculation in filter-based Forster resonance energy transfer microscopy for more accurate results. JOURNAL OF BIOMEDICAL OPTICS, 15(2). doi:10.1117/1.3407655

Li, H., Yu, P., Sun, Y., Felder, R. A., Periasamy, A., & Jose, P. A. (2010). Actin cytoskeleton-dependent Rab GTPase-regulated angiotensin type I receptor lysosomal degradation studied by fluorescence lifetime imaging microscopy. JOURNAL OF BIOMEDICAL OPTICS, 15(5). doi:10.1117/1.3484751