HoloMonitor®
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Catalog excerpts

HoloMonitor® - 1

Imaging living cells without compromising cell integrity Label-free live cell imaging and analysis for cell population studies by tracking and quantifying individual cells over time — a technical explanation. The HoloMonitor® label-free live cell imaging system is based on the principle of quantitative phase imaging, enabling non-invasive visualization and quanti cation of living cells without compro ising cell integrity. fi m Here we describe the rationale and advantages of using quanti ative phase imaging for live cell kinetic t analysis of cellular events — explaining the power of live cell time-lapse cytometry and how cells are made visible without labels or stains. Peter Egelberg CEO & Founder, Phase Holographic Im

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LABEL-FREE LIVE CELL IMAGING — HOW DOES IT WORK? Just like water waves, light waves of a specific wavelength have two principal charac erstics: amplitude and t i phase. Amplitude corre ponds to light intensity and is the height of the wave, measured from crest to trough. s Phase describes whether a wave is currently at its crest, in its trough, or some here in between. w Invisible cells For a living cell to be visible to the naked eye or in a light micro scope, the light arriving from the cell must differ in intensity from the back round, i.e. differ in amplitude. Unfortunately, living...

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THE SOLUTION With computer technology, it is possible both to quan fy and visualize phase-shifts. This tech ique is called ti n quantitative phase imaging (QPI) or quanti ative phase contrast microscopy to distinguish it from con entional t v phase contrast microscopy. QPI provides both quantitative and beautiful images of living cells, transforming phase micro copy and live cell imaging into a quantitative tool for detailed cell analysis. s An example of a quantitative phase image of cells in 3D, created by HoloMonitor. The height of the cell and its color tone corre pond to the optical...

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Simpler live cell identification As the cells are well separated from the background in a quanti tative phase image, simpler and more robust computer algo rithms can be used to identify individual cells in such images. Optical thickness Unlike in phase contrast microscopy images, the inten ity of a pixel s in a phase image has a direct physical meaning. It corresponds to the optical thickness of the cell, which is the physical height of the cell multiplied by the optical density of the cell at that point. Con e uently, cell structures that are optically dense like s q lipid droplets will...

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HOLOGRAPHIC MICROSCOPY There are several forms of quanti ative phase micro copy. The HoloMonitor® live cell time-lapse cytometers1 t s employ the most common form, holographic micro copy. s Parallel laser light Diode laser Reference beam Sample beam Cell culture Imprinted laser light Image sensor Shallow water imprinted beach waves Holographic microscopy creates quantitative phase images by letting a sample beam and a reference beam interfere to create an interference pattern or hologram, as shown above. The hologram is recorded by an image sensor and computer processed to produce the phase...

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An example time-lapse image sequence of a monster HeLa cell, imaged by HoloMonitor. HeLa cells are cancer cells named after Henrietta Lacks, who in the early 1950s donated the first cells that was successfully kept alive and cultured in a laboratory environment. HeLa cells and other immortal cells are today routinely cultured by scientists to study the complex behavior of cells and their response to drug treatments. In addition to identifying each individual cell, HoloMonitor provides data for analysis of more than 30 morphological parameters. However, the true power of time-lapse cytometry...

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When HoloMonitor was compared with standard transwell migration and invasion assays, the tracking of non-directional motility of individual cells over time produced highly correlated relative motilities. Evaluation of holographic imaging cytometer HoloMonitor® M4 motility applications, Cytometry (2018) — Yuntian Zhang and Robert L. Judson, University of California, San Francisco LIVE CELL IMAGING & ANALYSIS ASSAYS Altogether the described features enable a range of biologically relevant parameters to be effortlessly anayzed l over time. The label-free live cell imaging and analysis...

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REFERENCES 1. Cells and Holograms – Holograms and Digital Holographic Microscopy as a Tool to Study the Morphology of Living Cells, K. Alm, Z. El-Schich, M. Falck Miniotis, A. Gjörloff Wingren, B. Janicke and S. Oredsson, Holography - Basic Principles and Contemporary Applications (2013). 2. Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy, Z. El‑Schich, A. Leida Mölder and A. Gjörloff Wingren, Appl. Sci. (2018). 3. Evaluation of Holographic Imaging Cytometer HoloMonitor M4® Motility Applications, Y. Zhang and R. L. Judson, Cytometry...

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