Q-PHASE: Quantitative, label-free imaging cytometry

Q-PHASE: Quantitative, label-free imaging cytometry
Q-PHASE Multimodal Holographic Microscope

OBSERVE cells with unmatched resolution

Q-PHASE microscope has been designed to image the cells with unmatched clarity and without any labelling. A unique patented setup of QPI, allows clearly detecting cellular boundaries and mass changes inside the cells.

Unparalleled cellular contrast

The quantitative phase imaging is directly dependent on cellular mass (thickness & refractive index).

Transparent cells made visible

As the QPI detects even the slightest changes in the cellular mass, even the most transparent cells and their parts can be distinguished from the background.

Look inside the cells

Extraordinary mass detection of QPI allows also seeing changes in internal parts of the cells, such as nuclei, vacuoles, and many more, again, without any labelling.

Automatically SEGMENT the data with high accuracy

High clarity data based on precise image background compensation allows quick segmentation and identification of cellular boundaries of individual moving cells in large populations.
Automated segmentation of QPI image

ANALYSE quantitative data for every cell

Automated segmentation - The excellent sofware segmentation capability allows obtaining data on the fly with excellent accuracy.
Segmented image ready for subsequent data mining
Automated cellular data analysis using Cell Analyser module

TRACK cell dynamics over long time periods

We have developed an automatic label-free method to track the dynamic phenotype of thousands of single cells in heterogeneous populations over long time periods.

QPI Time-lapse of cell interactions. (A) Time-lapse imaging of entosis
Cell tracking during entosis

Get the DATA from single cells to large populations

Quantitative analysis for each cell - This approach efficiently uncovers and classifies cellular subtypes as well as discovers rare cells from large cellular populations.
Autophagic human ovarian cancer cell line A2780 (fluorescence staining: Hoechst 33342, CytoID) (20x mag.)
Human ovarian cancer cell line A2780 (fluorescence staining: Hoechst 33342, CytoID) (4x mag.)

New look at cell biology: Applications

  • Cell differentiation
  • Cancer cell biology
  • Biocompatibility
  • Drug testing
  • Cell death analysis
  • Cell cycle analysis
  • Plant biology
  • Hydrobiology
  • Cells in 3D environment

Metastatic cancer cell – immune cell interaction (T-cell invading polyploid cancer cell)

Formation of polyploid ovarian cancer cell A2780

Human embryonal stem cells: Colony formation

Diversity of metastatic cancer cells population (PC-3 cells)

Communication of human chondrogenic cells

Sperm cells movement for semen analysis

Intake of apodox and treatment response of human breast cancer cell line MDA-MB-231

Intercellular communication of human metastatic polyploid prostate cancer cell PC-3

Observation of HEK293 cell population transfected with GFP-Vesicular Stomatis Virus Coat Protein

5 days of human embryonal stem cells (HESC) colony life

Related Application Notes

Quantitative Phase Imaging (QPI) provided by Q-PHASE – TESCAN multimodal holographic microscope – allows observation of cells reactions to different treatments without any other added dyes. Combination of QPI with fluorescence provides the possibility to observe cell processes with low phototoxicity and simultaneously verify observed processes using a single instrument. This innovative approach opens interesting opportunities in cancer research. Through experiments, researchers look at how cancer cells behave and try to understand cancer at its deepest levels. Understanding the basic processes of these cells can help researchers figure out what controls cell division and cell death; find out what makes cancer cells spread or metastasize, identify unique characteristics of cancer cells to design newtherapeutic strategies, and, find out why certain cancer cells become resistant to therapy. In some of these experiments, cells taken from tumors of people with cancer are studied.
pdf – 1.3 MB
Q-PHASE Live Cell Imaging
Q-PHASE, the multimodal holographic microscope, is a unique instrument for quantitative phase imaging (QPI). The main application of this technique is in live cell imaging where advantages such as no need for labeling, low phototoxicity, easy segmentation, cell dry mass interpretation of measured signal and suitability for long term experiments are used. Q-PHASE is built as a transmitted light microscope in an inverted configuration for easy handling with biological samples. Appropriate conditions for live cells are ensured by the microscope incubator heated to 37°C and low exposures of light for QPI. Moreover, there is no need for specific sample preparation. The cells are just seeded into a suitable observation chamber and examined.
pdf – 1.7 MB

Key features

  • Simple Automated Image segmentation and processing – comparable to fluorescence data processing
  • No image artifacts such as halo effect (as opposed to techniques based on Zernike phase contrast illumination)
  • Enables very precise detection of cell boundaries
  • Label-free – no staining is needed, simple sample preparation, observation of live cells in their native environment, no photobleaching problems
  • Low phototoxicity – low light power density (107 × lower than fluorescence microscopy) allows long-term observations (for days)
  • Coherence-gating effect – Q-PHASE special feature enabling to observe samples even in scattering media (phospholipid emulsions, extracellular matrices, etc.)
  • Multimodality – fully integrated fluorescence module, simulated DIC and brightfield which enables automatic multimodal imaging of the sample
  • High-quality QPI – unique Q-PHASE’s optical setup allows using incoherent illumination which provides extraordinary imaging quality without any compromises
  • Strong suppression of coherent noise (speckles) & parasitic interferences (as opposed to laser-based approaches)
  • ‚‚Lateral resolution of conventional microscopes (up to 2× better when compared to common laser-based approaches or pinhole spatial filtering based techniques)
  • Fast acquisition – the use of off-axis holographic approach makes Q-PHASE a single-shot instrument, thus enabling imaging of very fast cell dynamics
  • Full motorization – focusing, sample stage, objective exchange, fluorescence filters
  • Automated multidimensional acquisition – time-lapse, channel, position, Z-stack
  • Quantitative – phase values can be recalculated e.g. to cell dry-mass density (pg/μm2) or direct topography with nanometer sensitivity (usually non-biological samples with homogeneous refractive index distribution)
  • High phase detection sensitivity – enables to detect even the smallest changes in axial direction, very sensitive detection of morphology or position changes

Principles of quantitative phase imaging

The time of propagation of light in a specific environment depends on the refractive index as well as the distance of the optical path. Therefore, when a light wave travels through a sample with varying refractive index and/or height, its wavefront is distorted and a change in the phase distribution of this wave occurs. The Q-PHASE allows to detect the phase distribution in the sample plane. This process of phase detection at a sample plane is usually referred to as quantitative phase imaging. Quantitative phase image can provide information on sample morphology, topography or cell dry-mass distribution. Cell dry mass is quantified in pg/μm2 and can be calculated directly from phase values detected in each pixel. Quantitative phase imaging provides very simple and sensitive way for monitoring of cell reactions to treatment and analyses of movement, growth, area, shape and many other parameters. Various color LUTs are often used for representation of phase images to easily distinguish different phase values.

Patented optical setup

The Q-PHASE microscope consists of two arms, object arm and reference arm. The arms have similar microscope setups with a common illumination system. The sample is placed into the object arm, and the so-called reference sample (blank) is placed into the reference arm. The beams in each arm pass through the inserted sample and are combined at the image plane of the microscope. Thanks to the Q-PHASE’s unique patented optical setup, the beams interfere and form a hologram even when illuminated with a halogen lamp or a LED. The hologram is then recorded by a detector and a quantitative phase image is extracted from the hologram in real time by a computer.

Product Brochure

Q-PHASE Brochure
Q-PHASE - MULTIMODAL HOLOGRAPHIC MICROSCOPE - Quantitative phase imaging. Download the brochure!
pdf – 891 kB

Q-PHASE in Springer Nature

New article dedicated to the Q-PHASE is available on SHAREDIT by Springer Nature. Name of article is "Holography microscopy as an artifact-free alternative to phase-contrast". 

Holography microscopy (HM) is one of the quantitative phase imaging techniques, which has been finding new applications in life science, especially in morphological screening, cell migration, and cancer research. Holography microscopy imaging provides artefact-free data essential for subsequent image processing and analysis. This work gives evidence that holography microscopy represents a very convenient method that can be successfully applied to phenotypic screening using image analysis implemented in a high-throughput workflow.