Prof. Rohit Bhargava Profile

Prof. Rohit Bhargava

Professor at Beckman Institute for Advanced Science & Technology

SPIE Involvement:

Conference Program Committee | Author

Publications (33)

SPIE Journal Paper | 16 July 2024 Open Access

Capturing cell morphology dynamics with high temporal resolution using single-shot quantitative phase gradient imaging

Sun Woong Hur, MinSung Kwon, Revathi Manoharaan, Melika Haji Mohammadi, Ashok Zachariah Samuel, Michael Mulligan, Paul Hergenrother, Rohit Bhargava

JBO, Vol. 29, Issue S2, S22712, (July 2024) https://doi.org/10.1117/12.10.1117/1.JBO.29.S2.S22712

KEYWORDS: Cell death, Polarization, Microscopes, Tunable filters, Temporal resolution, Fluorescence, Phase reconstruction, Image restoration, Fluorescence imaging, Digital image correlation

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SignificanceLabel-free quantitative phase imaging can potentially measure cellular dynamics with minimal perturbation, motivating efforts to develop faster and more sensitive instrumentation. We characterize fast, single-shot quantitative phase gradient microscopy (ss-QPGM) that simultaneously acquires multiple polarization components required to reconstruct phase images. We integrate a computationally efficient least squares algorithm to provide real-time, video-rate imaging (up to 75 frames/s). The developed instrument was used to observe changes in cellular morphology and correlate these to molecular measures commonly obtained by staining.AimWe aim to characterize a fast approach to ss-QPGM and record morphological changes in single-cell phase images. We also correlate these with biochemical changes indicating cell death using concurrently acquired fluorescence images.ApproachHere, we examine nutrient deprivation and anticancer drug-induced cell death in two different breast cell lines, viz., M2 and MCF7. Our approach involves in-line measurements of ss-QPGM and fluorescence imaging of the cells biochemically labeled for viability.ResultsWe validate the accuracy of the phase measurement using a USAF1951 pattern phase target. The ss-QPGM system resolves 912.3 lp/mm, and our analysis scheme accurately retrieves the phase with a high correlation coefficient (∼0.99), as measured by calibrated sample thicknesses. Analyzing the contrast in phase, we estimate the spatial resolution achievable to be 0.55 μm for this microscope. ss-QPGM time-lapse live-cell imaging reveals multiple intracellular and morphological changes during biochemically induced cell death. Inferences from co-registered images of quantitative phase and fluorescence suggest the possibility of necrosis, which agrees with previous findings.ConclusionsLabel-free ss-QPGM with high-temporal resolution and high spatial fidelity is demonstrated. Its application for monitoring dynamic changes in live cells offers promising prospects.

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Proceedings Article | 13 March 2024 Presentation + Paper

Metal carbonyl loaded nanoparticles for breast cancer biomarker detection

Craig Richard, Pei-Hsuan Hsieh, Matthew Confer, Rohit Bhargava

Proceedings Volume 12858, 1285805 (2024) https://doi.org/10.1117/12.3002618

KEYWORDS: Nanoprobes, Nanoparticles, Infrared imaging, Infrared spectroscopy, Polystyrene, FT-IR spectroscopy, Breast cancer, Biomolecules

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Proceedings Article | 13 March 2024 Poster

Characterization of a laser-scanning mid-infrared microscope with digitally referenced per-pulse detection

Ruo-Jing Ho, Kevin Yeh, Yen-Ting Liu, Matthew Confer, Rohit Bhargava

Proceedings Volume PC12853, PC128530T (2024) https://doi.org/10.1117/12.2692392

KEYWORDS: Quantum cascade lasers, Mid-IR, Microscopes, Quantum detection, Vibration, Thermography, Thermal stability, Spectrometers, Quantum hardware, Pulsed laser operation

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Proceedings Article | 13 March 2024 Presentation

Development and translation of an infrared laser scanning microscope for whole-slide chemical imaging

Kevin Yeh, Ishaan Sharma, Kianoush Falahkheirkhah, Matthew Confer, Andres Orr, Yen-Ting Liu, Ruo-Jing Ho, Mykolas Tamonis, John Cheville, Rohit Bhargava

Proceedings Volume PC12855, PC128551A (2024) https://doi.org/10.1117/12.3003524

KEYWORDS: Imaging spectroscopy, Microscopes, Infrared spectroscopy, Infrared lasers, Biomedical optics, Chemical analysis, Spatial resolution, Quantum imaging, Quantum frequencies, Quantum cascade lasers

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Proceedings Article | 13 March 2024 Presentation

Null-deflection scanning probe infrared (NDIR) spectroscopic imaging: from analytical models to applications

Anirudha Rao, Seth Kenkel, Rohit Bhargava

Proceedings Volume PC12839, PC128390U (2024) https://doi.org/10.1117/12.3008670

KEYWORDS: Biological samples, Imaging spectroscopy, Resonance enhancement, Signal to noise ratio, Biological imaging, Spectroscopy, Spatial resolution, Image quality, Visualization, Molecules

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Proceedings Article | 6 March 2023 Presentation

Uncovering new utility in infrared spectroscopic imaging by pushing the limits of accuracy, speed, and resolution

Rohit Bhargava, Kevin Yeh, Seth Kenkel, Yamuna Dilip Phal, Ruo-Jing Ho, Kianoush Falahkheirkhah

Proceedings Volume PC12373, PC1237303 (2023) https://doi.org/10.1117/12.2659474

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Proceedings Article | 8 March 2021 Presentation

Infrared chemical imaging with scanning quantum cascade laser microscopy systems

Kevin Yeh, Yamuna Phal, Rohit Bhargava

Proceedings Volume 11656, 116561A (2021) https://doi.org/10.1117/12.2578981

KEYWORDS: Imaging spectroscopy, Quantum cascade lasers, Infrared radiation, Infrared imaging, Imaging systems, Microscopy, Laser systems engineering, Tissues, Spectroscopes, Infrared spectroscopy

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Proceedings Article | 8 March 2021 Presentation

Mid-IR laser-based polarimetric imaging for polymeric and biological applications

Yamuna Phal, Kevin Yeh, Rohit Bhargava

Proceedings Volume 11656, 1165619 (2021) https://doi.org/10.1117/12.2579099

KEYWORDS: Polymers, Polarimetry, Mid-IR, Laser applications, Quantum cascade lasers, Infrared imaging, Microscopes, Tissues, Spectral resolution, Polarization

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Proceedings Article | 8 March 2021 Presentation

Pushing the limits of spatial and temporal capability of infrared spectroscopic imaging: a theory-based approach

Rohit Bhargava

Proceedings Volume 11656, 1165615 (2021) https://doi.org/10.1117/12.2579135

KEYWORDS: Infrared spectroscopy, Infrared imaging, Imaging spectroscopy, Infrared radiation, Spectroscopy, Microscopes, Infrared microscopy, Image processing, Upconversion, Spectroscopes

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Proceedings Article | 5 March 2021 Presentation

Statistical considerations in spectral histopathology and performance evaluation

Shachi Mittal, Rohit Bhargava

Proceedings Volume 11656, 1165614 (2021) https://doi.org/10.1117/12.2577920

KEYWORDS: Statistical analysis, Error analysis, Spectroscopy, Pathology, Infrared spectroscopy, Infrared imaging, Imaging spectroscopy, Data modeling, Tissue optics, Thermal modeling

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Proceedings Article | 5 March 2021 Presentation

Infrared chemical imaging for histologic interpretation of renal tissue

Ghazal Azarfar, Kianoush Falahkheirkhah, Meera Gopu, Jennifer Pfister, Sergey Brodsky, Tibor Nadasdy, Georgina Cheng, Anjali Satoskar, Rohit Bhargava

Proceedings Volume 11656, 1165612 (2021) https://doi.org/10.1117/12.2577337

KEYWORDS: FT-IR spectroscopy, Biopsy, Kidney, Neural networks, Hyperspectral imaging, Pathology, Tissues, Thermal modeling, Spectral resolution, Medicine

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Proceedings Article | 5 March 2020 Paper

Polarimetric infrared spectroscopic imaging using quantum cascade lasers

Yamuna Phal, Kevin Yeh, Rohit Bhargava

Proceedings Volume 11252, 1125210 (2020) https://doi.org/10.1117/12.2544392

KEYWORDS: Infrared imaging, Polarization, Infrared spectroscopy, Tissues, Collagen, Quantum cascade lasers, Imaging spectroscopy, Polarimetry, Infrared microscopy, Spectroscopy

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Proceedings Article | 15 March 2019 Paper

Multi-space-enabled deep learning of breast tumors improves prediction of distant recurrence risk

Dooman Arefan, Bingjie Zheng, David Dabbs, Rohit Bhargava, Shandong Wu

Proceedings Volume 10954, 109540L (2019) https://doi.org/10.1117/12.2513013

KEYWORDS: Tumors, Data modeling, Breast, Breast cancer, Mammography, Performance modeling, Statistical modeling, Cancer

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Proceedings Article | 5 March 2019 Paper

Nanoscale imaging of biological samples with responsivity corrected Atomic Force Microscopy-Infrared (AFM-IR) spectroscopy

Seth Kenkel, Rohit Bhargava

Proceedings Volume 10891, 108910A (2019) https://doi.org/10.1117/12.2510131

KEYWORDS: Infrared imaging, Imaging spectroscopy, Quantum cascade lasers, Spectroscopy, Infrared spectroscopy, Near field scanning optical microscopy, Infrared microscopy, Absorption

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Proceedings Article | 8 May 2018 Paper

Agile optoelectronic fiber sources for hyperspectral chemical sensing from SWIR to LWIR

Inuk Kang, Andrew Grant, Mihaela Dinu, James Jaques, Luke Pfister, Rohit Bhargava, P. Scott Carney

Proceedings Volume 10644, 106441E (2018) https://doi.org/10.1117/12.2305120

KEYWORDS: Long wavelength infrared, Biological and chemical sensing, Short wave infrared radiation, Spectroscopy, Standoff detection, Solitons, Detection and tracking algorithms

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Proceedings Article | 23 March 2016 Paper

A four class model for digital breast histopathology using high-definition Fourier transform infrared (FT-IR) spectroscopic imaging

Shachi Mittal, Tomasz Wrobel, L. Leslie, Andre Kadjacsy-Balla, Rohit Bhargava

Proceedings Volume 9791, 979118 (2016) https://doi.org/10.1117/12.2217358

KEYWORDS: FT-IR spectroscopy, Breast, Imaging spectroscopy, Spectroscopy, Pathology, Detection and tracking algorithms, Breast cancer, Tissue optics, Data analysis, Solid modeling, Tissues, Tumor growth modeling, Thermal modeling

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Proceedings Article | 23 March 2016 Paper

High-definition Fourier transform infrared spectroscopic imaging of prostate tissue

Tomasz Wrobel, Jin Tae Kwak, Andre Kadjacsy-Balla, Rohit Bhargava

Proceedings Volume 9791, 97911D (2016) https://doi.org/10.1117/12.2217341

KEYWORDS: FT-IR spectroscopy, Tissues, Imaging spectroscopy, Spectroscopy, Infrared imaging, Pathology, Fourier spectroscopy, Prostate, Tissue optics, Gold, Infrared spectroscopy, Infrared radiation, Data modeling

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Proceedings Article | 7 March 2016 Paper

Translation of infrared chemical imaging for cardiovascular evaluation

Saumya Tiwari, Jai Raman, Vijaya Reddy, Miranda Dawson, Rohit Bhargava

Proceedings Volume 9704, 97040X (2016) https://doi.org/10.1117/12.2230004

KEYWORDS: FT-IR spectroscopy, Imaging spectroscopy, Infrared imaging, Infrared spectroscopy, Infrared radiation, Spectroscopy, Tissues, Quantum cascade lasers, Biological research, Pathology

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Proceedings Article | 7 March 2016 Paper

Discrete frequency infrared imaging using quantum cascade lasers for biological tissue analysis

Kevin Yeh, Rohit Bhargava

Proceedings Volume 9704, 970406 (2016) https://doi.org/10.1117/12.2230003

KEYWORDS: Infrared imaging, Infrared spectroscopy, Quantum cascade lasers, Tissue optics, Biological research, Chemical analysis, Spectroscopy, Imaging spectroscopy, Infrared microscopy, Infrared radiation, Tissues, Absorbance, Signal to noise ratio, Absorption, Data acquisition, Confocal microscopy

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Proceedings Article | 19 March 2015 Paper

High-definition Fourier transform infrared spectroscopic imaging of breast tissue

L. Suzanne Leslie, Andre Kadjacsy-Balla, Rohit Bhargava

Proceedings Volume 9420, 94200I (2015) https://doi.org/10.1117/12.2082461

KEYWORDS: Infrared imaging, FT-IR spectroscopy, Imaging spectroscopy, Breast, Tissue optics, Image quality, Signal to noise ratio, Spectroscopy, Microscopes, Spectroscopes

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Proceedings Article | 17 March 2015 Paper

Fourier transform infrared (FT-IR) spectroscopy and imaging of the nucleus to characterize DNA contributions in different phases of the cell cycle

Saumya Tiwari, Xinying Zong, Sarah Holton, K. Prasanth, Rohit Bhargava

Proceedings Volume 9417, 941720 (2015) https://doi.org/10.1117/12.2082347

KEYWORDS: Infrared spectroscopy, FT-IR spectroscopy, Imaging spectroscopy, Spectroscopy, Proteins, Tissues, Infrared imaging, Packaging, Scattering, Infrared radiation

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Proceedings Article | 20 March 2014 Paper

Breast histopathology using random decision forests-based classification of infrared spectroscopic imaging data

David Mayerich, Michael Walsh, Andre Kadjacsy-Balla, Shachi Mittal, Rohit Bhargava

Proceedings Volume 9041, 904107 (2014) https://doi.org/10.1117/12.2043783

KEYWORDS: Tissues, FT-IR spectroscopy, Collagen, Infrared spectroscopy, Spectroscopy, Breast cancer, Imaging spectroscopy, Mid-IR, Biopsy, Breast

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Proceedings Article | 29 May 2013 Paper

Mid-infrared microspectroscopic imaging with a quantum cascade laser

Kevin Yeh, Matthew Schulmerich, Rohit Bhargava

Proceedings Volume 8726, 87260E (2013) https://doi.org/10.1117/12.2015984

KEYWORDS: Quantum cascade lasers, FT-IR spectroscopy, Mid-IR, Absorbance, Staring arrays, Imaging systems, Imaging spectroscopy, Infrared spectroscopy, Spectroscopes, Infrared imaging

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Proceedings Article | 10 February 2012 Open Access

Denoising and deblurring of Fourier transform infrared spectroscopic imaging data

Tan Nguyen, Rohith Reddy, Michael Walsh, Matthew Schulmerich, Gabriel Popescu, Minh Do, Rohit Bhargava

Proceedings Volume 8296, 82960M (2012) https://doi.org/10.1117/12.921144

KEYWORDS: FT-IR spectroscopy, Absorbance, Data modeling, Imaging spectroscopy, Point spread functions, Spectroscopy, Denoising, Sensors, Image processing, Imaging systems

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Proceedings Article | 1 February 2012 Paper

High-resolution mid-infrared imaging for disease diagnosis

Michael Walsh, David Mayerich, Andre Kajdacsy-Balla, Rohit Bhargava

Proceedings Volume 8219, 82190R (2012) https://doi.org/10.1117/12.909339

KEYWORDS: Tissues, FT-IR spectroscopy, Colon, Spatial resolution, Infrared imaging, Attenuated total reflectance, Imaging systems, Imaging spectroscopy, Image resolution, Absorbance

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Proceedings Article | 12 February 2011 Paper

Spatial-domain low-coherence quantitative phase microscopy for cancer diagnosis

Pin Wang, Rajan Bista, Rohit Bhargava, Randall Brand, Yang Liu

Proceedings Volume 7889, 78891Q (2011) https://doi.org/10.1117/12.873620

KEYWORDS: Cancer, Refractive index, Microscopy, Tissues, Scattering, Light sources, Signal detection, Light scattering, Breast cancer, Phase measurement

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Proceedings Article | 3 March 2009 Paper

Computer-aided detection of HER2 amplification status using FISH images: a preliminary study

Bin Zheng, Xiao-Hui Wang, Urvashi Surti, Rohit Bhargava, David Gur

Proceedings Volume 7260, 726028 (2009) https://doi.org/10.1117/12.810155

KEYWORDS: Image segmentation, Signal detection, Computer aided design, Breast cancer, Image analysis, Statistical analysis, Luminescence, Computer aided diagnosis and therapy, Genetics

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The amplification status of human epidermal growth factor receptors 2 (HER2) genes is strongly associated with clinical outcome in patients with breast cancer. The American Society of Clinical Oncology Tumor Marker Guidelines Panel has recommended routine testing of HER2 status on all newly diagnosed metastatic breast cancers since 2001. Although fluorescent in situ hybridization (FISH) technology provides superior accuracy as compared with other approaches, current manual FISH analysis methods are somewhat subjective, tedious, and may introduce interreader variability. The goal of this preliminary study is to develop and test a computer-aided detection (CAD) scheme to assess HER2 status using FISH images. Forty FISH images were selected for this study from our genetic laboratory. The CAD scheme first applies an adaptive, iterative threshold method followed by a labeling algorithm to segment cells of possible interest. A set of classification rules is then used to identify analyzable interphase cells and discard nonanalyzable cells due to cell overlapping and/or other image staining debris (or artifacts). The scheme then maps the detected analyzable cells onto two other gray scale images corresponding to the red and green color of the original image followed by application of a raster scan and labeling algorithms to separately detect the HER-2/neu ("red") and CEP17 ("green") FISH signals. A simple distance based criterion is applied to detect and merge split FISH signals within each cell. The CAD scheme computes the ratio between independent "red" and "green" FISH signals of all analyzable cells identified on an image. If the ratio is ≥ 2.0, the FISH image is assumed to have been acquired from a HER2+ case; otherwise, the FISH image is assumed to have been acquired from HER2- case. When we applied the CAD scheme to the testing dataset, the average computed HER2 amplification ratios were 1.06±0.25 and 2.53±0.81 for HER2- and HER2+ samples, respectively. The results show that the CAD scheme has the ability to automatically detect HER2 status using FISH images. The success of CAD-guided FISH image analysis could result in a more objective, consistent, and efficient approach in determining HER2 status of breast cancers.

Proceedings Article | 24 February 2009 Paper

Spectral histology of breast tissue using mid-infrared spectroscopic imaging

F. Nell Pounder, Rohit Bhargava

Proceedings Volume 7182, 718206 (2009) https://doi.org/10.1117/12.810125

KEYWORDS: Tissues, FT-IR spectroscopy, Breast, Imaging spectroscopy, Image classification, Tissue optics, Spectroscopy, Blood vessels, Image segmentation, Tumors

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Proceedings Article | 24 February 2009 Paper

Histologic models for optical tomography and spectroscopy of tissues

Rohit Bhargava, Brynmor Davis

Proceedings Volume 7174, 71742H (2009) https://doi.org/10.1117/12.810119

KEYWORDS: Tissues, Optical coherence tomography, Spectroscopy, FT-IR spectroscopy, Imaging spectroscopy, Absorption, Signal to noise ratio, Scattering, Statistical modeling, Optical spectroscopy

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Proceedings Article | 20 February 2009 Paper

Validating the cancer diagnosis potential of mid-infrared spectroscopic imaging

F. Nell Pounder, Rohith Reddy, Michael Walsh, Rohit Bhargava

Proceedings Volume 7186, 71860F (2009) https://doi.org/10.1117/12.810122

KEYWORDS: Tissues, FT-IR spectroscopy, Breast, Sensors, Tumors, Image segmentation, Spectral resolution, Cancer, Spectroscopy, Imaging spectroscopy

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Proceedings Article | 26 August 2008 Paper

Tunable multilayered nanospheres as probes for surface-enhanced Raman spectroscopy

Anil Kodali, Rohit Bhargava

Proceedings Volume 7032, 70320V (2008) https://doi.org/10.1117/12.798168

KEYWORDS: Dielectrics, Gold, Metals, Silica, Raman spectroscopy, Refractive index, Optical spheres, Multilayers, Silver, Raman scattering

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Proceedings Article | 21 February 2008 Paper

Structural and biochemical characterization of engineered tissue using FTIR spectroscopic imaging: melanoma progression as an example

Rohit Bhargava, Rong Kong

Proceedings Volume 6870, 687004 (2008) https://doi.org/10.1117/12.762511

KEYWORDS: Skin, Tissues, Tumors, FT-IR spectroscopy, Imaging spectroscopy, Spectroscopy, Melanoma, Tissue optics, Systems modeling, Infrared imaging

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Proceedings Article | 8 February 2008 Paper

Practical protocols for fast histopathology by Fourier transform infrared spectroscopic imaging

Frances Keith, Rohith Reddy, Rohit Bhargava

Proceedings Volume 6853, 685306 (2008) https://doi.org/10.1117/12.762468

KEYWORDS: Tissues, FT-IR spectroscopy, Spectral resolution, Signal to noise ratio, Data acquisition, Image classification, Tissue optics, Image segmentation, Breast, Imaging spectroscopy

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Showing 5 of 33 publications

Conference Committee Involvement (26)

Digital and Computational Pathology

16 February 2025 | San Diego, California, United States

Advanced Chemical Microscopy for Life Science and Translational Medicine 2025

25 January 2025 | San Francisco, California, United States

Digital and Computational Pathology

19 February 2024 | San Diego, California, United States

Biomedical Vibrational Spectroscopy 2024: Advances in Research and Industry

27 January 2024 | San Francisco, California, United States

Advanced Chemical Microscopy for Life Science and Translational Medicine 2024

27 January 2024 | San Francisco, California, United States

Digital and Computational Pathology

20 February 2023 | San Diego, California, United States

Advanced Chemical Microscopy for Life Science and Translational Medicine 2023

28 January 2023 | San Francisco, California, United States

Digital and Computational Pathology

20 February 2022 | San Diego, California, United States

Advanced Chemical Microscopy for Life Science and Translational Medicine 2022

23 January 2022 | San Francisco, California, United States

Biomedical Vibrational Spectroscopy 2022: Advances in Research and Industry

22 January 2022 | San Francisco, California, United States

Advanced Chemical Microscopy for Life Science and Translational Medicine 2021

6 March 2021 | Online Only, California, United States

Digital and Computational Pathology

15 February 2021 | Online Only, California, United States

Digital Pathology

19 February 2020 | Houston, Texas, United States

Biomedical Vibrational Spectroscopy 2020: Advances in Research and Industry

1 February 2020 | San Francisco, California, United States

Advanced Chemical Microscopy for Life Science and Translational Medicine

1 February 2020 | San Francisco, California, United States

Digital Pathology

20 February 2019 | San Diego, California, United States

Digital Pathology

11 February 2018 | Houston, Texas, United States

Biomedical Vibrational Spectroscopy 2018: Advances in Research and Industry

27 January 2018 | San Francisco, California, United States

Digital Pathology

12 February 2017 | Orlando, Florida, United States

Digital Pathology Posters

12 February 2017 | Orlando, FL, United States

Digital Pathology

2 March 2016 | San Diego, California, United States

Biomedical Vibrational Spectroscopy 2016: Advances in Research and Industry

13 February 2016 | San Francisco, California, United States

Digital Pathology

25 February 2015 | Orlando, Florida, United States

Digital Pathology

16 February 2014 | San Diego, California, United States

Biomedical Vibrational Spectroscopy VIII: Advances in Research and Industry

1 February 2014 | San Francisco, California, United States

Biomedical Vibrational Spectroscopy VII: Advances in Research and Industry

21 January 2012 | San Francisco, California, United States

Showing 5 of 26 Conference Committees

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