scholarly journals Epi-illumination gradient light interference microscopy for imaging opaque structures

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Mikhail E. Kandel ◽  
Chenfei Hu ◽  
Ghazal Naseri Kouzehgarani ◽  
Eunjung Min ◽  
Kathryn Michele Sullivan ◽  
...  
Keyword(s):  
Multiple Scattering ◽  
Imaging Modality ◽  
Single Cells ◽  
Scattering Problem ◽  
Biological Tissues ◽  
Interference Microscopy ◽  
Model Organisms ◽  
Phase Imaging ◽  
Label Free ◽  
Light Interference

Abstract Multiple scattering and absorption limit the depth at which biological tissues can be imaged with light. In thick unlabeled specimens, multiple scattering randomizes the phase of the field and absorption attenuates light that travels long optical paths. These obstacles limit the performance of transmission imaging. To mitigate these challenges, we developed an epi-illumination gradient light interference microscope (epi-GLIM) as a label-free phase imaging modality applicable to bulk or opaque samples. Epi-GLIM enables studying turbid structures that are hundreds of microns thick and otherwise opaque to transmitted light. We demonstrate this approach with a variety of man-made and biological samples that are incompatible with imaging in a transmission geometry: semiconductors wafers, specimens on opaque and birefringent substrates, cells in microplates, and bulk tissues. We demonstrate that the epi-GLIM data can be used to solve the inverse scattering problem and reconstruct the tomography of single cells and model organisms.

scholarly journals Quantitative Histopathology of Stained Tissues using Color Spatial Light Interference Microscopy (cSLIM)

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hassaan Majeed ◽  
Adib Keikhosravi ◽  
Mikhail E. Kandel ◽  
Tan H. Nguyen ◽  
Yuming Liu ◽  
...  
Keyword(s):  
New Technologies ◽  
Imaging Modality ◽  
Interference Microscopy ◽  
Phase Imaging ◽  
Bright Field Image ◽  
Interferometric Imaging ◽  
Breast Cancer Patients ◽  
Prognostic Information ◽  
Light Interference ◽  
Tissue Phase

Abstract Tissue biopsy evaluation in the clinic is in need of quantitative disease markers for diagnosis and, most importantly, prognosis. Among the new technologies, quantitative phase imaging (QPI) has demonstrated promise for histopathology because it reveals intrinsic tissue nanoarchitecture through the refractive index. However, a vast majority of past QPI investigations have relied on imaging unstained tissues, which disrupts the established specimen processing. Here we present color spatial light interference microscopy (cSLIM) as a new whole-slide imaging modality that performs interferometric imaging on stained tissue, with a color detector array. As a result, cSLIM yields in a single scan both the intrinsic tissue phase map and the standard color bright-field image, familiar to the pathologist. Our results on 196 breast cancer patients indicate that cSLIM can provide stain-independent prognostic information from the alignment of collagen fibers in the tumor microenvironment. The effects of staining on the tissue phase maps were corrected by a mathematical normalization. These characteristics are likely to reduce barriers to clinical translation for the new cSLIM technology.

scholarly journals Monitoring Reactivation of Latent HIV by Label-Free Gradient Light Interference Microscopy

2020 ◽  
Author(s):  
Neha Goswami ◽  
Yiyang Lu ◽  
Mikhail E. Kandel ◽  
Michael J. Fanous ◽  
Kathrin Bohn-Wippert ◽  
...  
Keyword(s):  
Mass Density ◽  
Single Cells ◽  
Dry Mass ◽  
Interference Microscopy ◽  
Label Free ◽  
Light Interference ◽  
Mean Diameter ◽  
Latently Infected ◽  
Infected Cells ◽  
Latent Hiv

SummaryLatent human immunodeficiency virus (HIV) reservoirs in infected individuals present the largest barrier to a cure. The first step towards overcoming this challenge is to understand the science behind latency-reactivation interplay. Fluorescence imaging of GFP-tagged HIV has been the main method for studying reactivation of latent HIV in individually infected cells. In this paper, we report insights provided by label-free, gradient light interference microscopy (GLIM) about the changes in measures including dry mass, diameter, and dry mass density associated with infected cells that occur upon reactivation. We discovered that mean cell dry mass and mean diameter of latently infected cells treated with reactivating drug, TNF-α, are higher for cells with reactivated HIV as compared to those with latent disease. Results also indicate that cells with mean dry mass and diameter less than 10pg and 8µm, respectively, remain exclusively in the latent state. Also, cells with mean dry mass greater than 23pg and mean diameter greater than 11µm have a higher probability of reactivating. This study is significant as it presents a new label-free approach to quantify latent reactivation of a virus in single cells based on changes in cell morphology.

scholarly journals Topography and refractometry of sperm cells using SLIM

2017 ◽  
Author(s):  
Lina Liu ◽  
Mikhail E. Kandel ◽  
Marcello Rubessa ◽  
Sierra Schreiber ◽  
Mathew Wheeler ◽  
...  
Keyword(s):  
Refractive Index ◽  
Interference Microscopy ◽  
Phase Imaging ◽  
Label Free ◽  
Quantitative Phase Imaging ◽  
Ensemble Averaging ◽  
Thickness Profile ◽  
Light Interference ◽  
Phase Sensitive

AbstractCharacterization of spermatozoon viability is a common test in treating infertility. Recently, it has been shown that label-free, phase-sensitive imaging can provide a valuable alternative for this type of assay. Here, we employ spatial light interference microscopy (SLIM) to decouple the thickness and refractive index information of individual cells. This procedure was enabled by quantitative phase imaging cells on media of two different refractive indices and using a numerical tool to remove the curvature from the cell tails. This way, we achieved ensemble averaging of topography and refractometry of 100 cells in each of the two groups. The results show that the thickness profile of the cell tail goes down to 150 nm and the refractive index can reach values of 1.6 close to the head.

scholarly journals Monitoring Reactivation of Latent HIV by Label-Free Gradient Light Interference Microscopy

iScience ◽  
2021 ◽  
pp. 102940
Author(s):  
Neha Goswami ◽  
Yiyang Lu ◽  
Mikhail E. Kandel ◽  
Michael J. Fanous ◽  
Kathrin Bohn-Wippert ◽  
...  
Keyword(s):  
Interference Microscopy ◽  
Label Free ◽  
Light Interference ◽  
Latent Hiv

scholarly journals Label-free, multi-scale imaging of ex-vivo mouse brain using spatial light interference microscopy

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Eunjung Min ◽  
Mikhail E. Kandel ◽  
CheMyong J Ko ◽  
Gabriel Popescu ◽  
Woonggyu Jung ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Ex Vivo ◽  
Interference Microscopy ◽  
Label Free ◽  
Multi Scale ◽  
Light Interference

scholarly journals Patterned substrates modulate growth and dynamics of 3D cellular systems

2019 ◽  
Author(s):  
Michael J. Fanous ◽  
Yanfen Li ◽  
Mikhail E. Kandel ◽  
Kristopher A. Kilian ◽  
Gabriel Popescu
Keyword(s):  
Cell Mass ◽  
Mouse Embryonic Fibroblast ◽  
Growth Patterns ◽  
Cellular Systems ◽  
Polar Coordinates ◽  
Interference Microscopy ◽  
Phase Imaging ◽  
Label Free ◽  
Patterned Substrates ◽  
Independent Manner

AbstractThe development of 3D cellular architectures during development and pathological processes involves intricate migratory patterns that are modulated by genetics and the surrounding microenvironment. The substrate composition of cell cultures has been demonstrated to influence growth, proliferation, and migration in 2D. Here we study the growth and dynamics of mouse embryonic fibroblast (MEF) cultures patterned in a tissue sheet which then exhibits 3D growth. Using gradient light interference microscopy (GLIM), a label-free quantitative phase imaging approach, we explored the influence of geometry on cell growth patterns and rotational dynamics. We apply, for the first time to our knowledge, dispersion-relation phase spectroscopy (DPS) in polar coordinates to generate the radial and rotational cell mass-transport. Our data show that cells cultured on engineered substrates undergo rotational transport in a radially independent manner and exhibit faster vertical growth than the control, unpatterned cells. The use of GLIM and polar DPS provides a novel quantitative approach to studying the effects of spatially patterned substrates on cell motility and growth.

scholarly journals Label-free screening of brain tissue myelin content using phase imaging with computational specificity (PICS)

2021 ◽  
Author(s):  
Michael Fanous ◽  
Chuqiao Shi ◽  
Megan Caputo ◽  
Laurie Rund ◽  
Rodney Johnson ◽  
...  
Keyword(s):  
Brain Tissue ◽  
Dry Mass ◽  
Phase Imaging ◽  
Label Free ◽  
Light Interference ◽  
Free Data ◽  
Highly Sensitive ◽  
Expert Pathologist ◽  
Molecular Specificity ◽  
The Central Nervous System

Inadequate myelination in the central nervous system is associated with neurodevelopmental complications. Thus, quantitative, high spatial resolution measurements of myelin levels are highly desirable. We used spatial light interference microcopy (SLIM), a highly sensitive quantitative phase imaging (QPI) technique, to correlate the dry mass content of myelin in piglet brain tissue with dietary changes and gestational size. We combined SLIM micrographs with an AI classifying model that allows us to discern subtle disparities in myelin distributions with high accuracy. This concept of combining QPI label-free data with AI for the purpose of extracting molecular specificity has recently been introduced by our laboratory as phase imaging with computational specificity (PICS). Training on nine thousand SLIM images of piglet brain tissue with the 71-layer transfer learning model Xception, we created a two-parameter classification to differentiate gestational size and diet type with an accuracy of 82% and 80%, respectively. To our knowledge, this type of evaluation is impossible to perform by an expert pathologist or other techniques.

scholarly journals Label-free imaging of collagen fibers in tissue slices using phase imaging with computational specificity

2021 ◽  
Author(s):  
Masayoshi Sakakura ◽  
Gabriel Popescu ◽  
Andre Kajdacsy-Balla ◽  
Virgilia Macias
Keyword(s):  
Second Harmonic ◽  
Imaging Modality ◽  
High Specificity ◽  
Ground Truth ◽  
Collagen Fibers ◽  
Nonlinear Susceptibility ◽  
Phase Imaging ◽  
Label Free ◽  
Tissue Slices ◽  
Disease Stratification

Evaluating the tissue collagen content in addition to the epithelial morphology has been proven to offer complementary information in histopathology, especially in disease stratification and patient survivability prediction. One imaging modality widely used for this purpose is second harmonic generation microscopy (SHGM), which reports on the nonlinear susceptibility associated with the collagen fibers. Another method is polarization light microscopy (PLM) combined with picrosirius-red (PSR) tissue staining. However, SHGM requires expensive equipment and provides limited throughput, while PLM and PSR staining are not part of the routine pathology workflow. Here, we advance phase imaging with computational specificity (PICS) to computationally infer the collagen distribution of unlabeled tissue, with high specificity. PICS utilizes deep learning to translate quantitative phase images (QPI) into corresponding PSR images with high accuracy and speed. Our results indicate that the distributions of collagen fiber orientation, length, and straightness reported by PICS closely match the ones from ground truth.

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