Label-free cell imaging

Introduction

QUANTITATIVE PHASE MICROSCOPY


Phasics offers a unique quantitative phase microscopy solution for label-free cell imaging. It delivers artifact-free images which enable accurately measuring valuable biological parameters for each single cell: morphology, dry mass…

cell-dry-mass-measurement

For

DRY MASS | at single cell level


New label-free biomarker

  • to quantitatively study cell events and pathologies: RBC disorder, cancer cell growth…
  • to monitor cell population over time: cell proliferation, stem cell differentiation, bioproduction…

 
Quantitative cell monitoring

MORPHOLOGY PARAMETERS
With high accuracy


  • Robust segmentation and measurement
  • Live cell tracking and cell motility measurement
  • Artefact-free images to observe intracellular components including cell membranes

Label-free-cell-segmentation

HIGH CONTENT IMAGING
Study each single cell of a large population


  • to run quantitative and statistical study of large cell population based on multiple parameter measurement: dry mass, surface, density…
  • Easy merging with other modalities to get even more data

 
subpopulation sorting

LABEL-FREE IMAGING
Non-invasive and fast preparation


  • Time-lapse microscopy over long period of time
  • No photobleaching

 
Intracellular imaging

Introduction

QUANTITATIVE PHASE IMAGE
| Meaningful pixel values


Each specimen introduces a delay in the light path when light propagates through it. This amount of delay (the phase shift) is locally measured by Phasics technique. An image is thus obtained for which each pixel value is the measured local phase shift.

This value is meaningful in cell biology as it directly relates to the specimen local thickness and composition (refractive index)

The sum of these pixels over the cell area is proven to give the dry mass of the cell, i.e. the weight without water.

Phase-shift-measurementOptical thickness = (n2-n1)xd

Set Up

Phasics technique enables label-free live microscopy without any change on the microscope. Thus it provides artifact-free images for accurate measurement and it is easy to integrate in very various experiments (time-lapse microscopy, high content screening, cell culture monitoring…)

Camera for quantitative phase microscopy
Phasics technique measures the phase shift thanks to a patented technology: the quadriwave lateral shearing interferometry. It consists in a camera-like instrument. It plugs on to any microscope. A dedicated software package computes the camera image in real time to deliver the quantitative phase image.

Advantages

HIGH CONTENT ANALYSIS
Phase, morphology and tracking data
for each single cell


  • Multiplex diagnostics combining phase shift parameters (homogeneity, density, dry mass…) and morphology data
  • Automated segmentation and analysis thanks to reliable phase shift measurement

LABEL-FREE
Easy & non-invasive


  • Fast assay preparation: no staining, no labelling
  • Non-invasive monitoring: long period time-lapse microscopy
  • Non-invasive cell classification and selection

PLUG & PLAY CAMERA
Any microscope – any modality


Suitable to:

  • Any microscope for versatile use in Research labs and imaging facilities
  • Any dishes, multi-well plates and any cell culture conditions
  • Incubator for long duration time-lapse

 
Easy set-up

  • Easy multi-modality
  • Possible OEM integration

REAL TIME
1 acquisition = 1 image


  • Applies to fast live cell event
  • No blur due to cell motion

ARTIFACT FREE
as the light is not filtered nor modified


  • Robust segmentation to enable automated analysis
  • Reliable high content study over large population
  • Phase noise < 0.5nm

Camera-like instrument for quantitative phase imaging

Introduction

Dry mass and morphometric parameters are proven indicators of many cell mechanisms:

  • cell viability (apoptose detection…)
  • cell growth (cell cycle status, , proliferation…)
  • cell differentiation
  • cell polarity
  • cell motility
  • cell abnormalities: shape change, heterogeneity, presence of parasite…

 
They are useful parameters for biology as shown in the following examples.

Example

BLOOD TESTING: pathology and disorder identification

Identify red blood cell blood disorder and blood-related pathology

Malaria diagnostic
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Anemia diagnostic
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Reticulocyte classification
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Label-free CELL POPULATION MONITORING

Get non-invasive live cell time-lapse for cell culture monitoring; cell selection in IVF or in regerative medicine (Stem cell, cornea cell)

In vitro fecondation
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Stem cell differentiation
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Cell culture monitoring
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DRUG TESTING & TOXICOLOGY

Quantify effects on cell mechanisms

Drug screening
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Cardiac cell
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Cell motility
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RESEARCH

Observe and measure cell-cell interaction or intracellular mechanisms

Neuron
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Adhesion fiber motility
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Cytoskeleton
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Bacteria proliferation
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Intracellular components
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Scientific publications

J. Biomed. Opt. 20(12), 126009 (Dec 22, 2015)

DOI: 10.1117/1.JBO.20.12.126009

Living cell dry mass measurement using quantitative phase imaging with quadriwave lateral shearing interferometry: an accuracy and sensitivity discussion

Applied Physics Letter 106, 233703

DOI: 10.1063/1.4922525

Phase imaging microscopy for the diagnostics of plasma-cell interaction

Press release

"Fifth edition of the FIEEC Applied Research Prize" (French press release)

The 2nd prize was given to Serge Monneret from Institut Carnot STAR to reward his collaboration with Phasics for quantitative phase microscopy"
French press release on FIEEC site

Biophysical Journal, 106,8 pp 1588-1595

DOI: 10.1016/j.bpj.2014.02.023

Fast label-free cytoskeletal network imaging in living mammalian cells

Journal of Biomedical Optics, SPIE

DOI: 10.1117/1.JBO.17.7.076004

Optical detection and measurement of living cell morphometric features with single-shot quantitative phase microscopy

Optics Express, Vol. 17, Issue 15, pp.13080-13094

DOI: 10.1364/OE.17.013080

Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells

Application Note

Cell dry mass to monitor single cell growth

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Red Blood Cell disorders classification using dry mass and morphology

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Malaria parasite counting using fast label-free protocol

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