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 ◽  
...  
2020 ◽  
Author(s):  
Neha Goswami ◽  
Yiyang Lu ◽  
Mikhail E. Kandel ◽  
Michael J. Fanous ◽  
Kathrin Bohn-Wippert ◽  
...  

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.


2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Eunjung Min ◽  
Mikhail E. Kandel ◽  
CheMyong J Ko ◽  
Gabriel Popescu ◽  
Woonggyu Jung ◽  
...  

2017 ◽  
Author(s):  
Lina Liu ◽  
Mikhail E. Kandel ◽  
Marcello Rubessa ◽  
Sierra Schreiber ◽  
Mathew Wheeler ◽  
...  

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.


2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Mikhail E. Kandel ◽  
Chenfei Hu ◽  
Ghazal Naseri Kouzehgarani ◽  
Eunjung Min ◽  
Kathryn Michele Sullivan ◽  
...  

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.


2011 ◽  
Vol 16 (2) ◽  
pp. 026019 ◽  
Author(s):  
Zhuo Wang ◽  
Larry Millet ◽  
Vincent Chan ◽  
Huafeng Ding ◽  
Martha U. Gillette ◽  
...  

2017 ◽  
Author(s):  
Hassaan Majeed ◽  
Tan Huu Nguyen ◽  
Mikhail Eugene Kandel ◽  
Andre Kajdacsy-Balla ◽  
Gabriel Popescu

Breast cancer is the most common type of cancer among women worldwide. The standard histopathology of breast tissue, the primary means of disease diagnosis, involves manual microscopic examination of stained tissue by a pathologist. Because this method relies on qualitative information, it can result in inter-observer variation. Furthermore, for difficult cases the pathologist often needs additional markers of malignancy to help in making a diagnosis. We present a quantitative method for label-free tissue screening using Spatial Light Interference Microscopy (SLIM). By extracting tissue markers of malignancy based on the nanostructure revealed by the optical path-length, our method provides an objective and potentially automatable method for rapidly flagging suspicious tissue. We demonstrated our method by imaging a tissue microarray comprising 68 different subjects - 34 with malignant and 34 with benign tissues. Three-fold cross validation results showed a sensitivity of 94% and specificity of 85% for detecting cancer. The quantitative biomarkers we extract provide a repeatable and objective basis for determining malignancy. Thus, these disease signatures can be automatically classified through machine learning packages, since our images do not vary from scan to scan or instrument to instrument, i.e., they represent intrinsic physical attributes of the sample, independent of staining quality.


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