In situ nanoscale observations of gypsum dissolution by digital holographic microscopy

The current invention concerns a method for detecting cancerous cells and/or classifying cells in a cell sample comprising the following steps: providing a cell sample; obtaining holographic information from said cell sample by digital holographic microscopy (DHM); deriving at least one cellular parameter from said holographic information, and; classifying said cells of cells sample; characterized in that said classification occurs by appointing a Scoring Factor to said cells of cell sample, based on said cellular parameters. In a second aspect, a system for the detection of cancerous cells and/or classification of cells in a cell sample is provided, employing the method as disclosed in the invention. In a final aspect, a method for updating and/or improving a database comprising thresholds linked to holographic information and the database related thereof is equally disclosed.

A method and system for detecting and/or classifying cancerous cells in a cell sample

Low cost digital holographic microscope for 3-D cell imaging by integrating smartphone and DVD optical head

Total internal reflection fluorescence (TIRF) microscopy benefits from high-sensitivity, low background noise, low photo-toxicity and high-contrast imaging of sub-cellular structures close to the membrane surface. Although, TIRF microscopy provides high-contrast imaging it does not provide quantitative information about morphological features of the biological cells. Here, we propose an integrated waveguide chip-based TIRF microscopy and label-free quantitative phase imaging (QPI). The evanescent field present on top of a waveguide surface is used to excite the fluorescence and an upright microscope is used to collect the signal. The upright microscope is converted into a Linnik-type interferometer to sequentially extract both the quantitative phase information and TIRF images of the cells. Waveguide chip-based TIRF microscopy benefits from decoupling of illumination and collection light path, large field of view imaging and pre-aligned configuration for multi-color TIRF imaging. The proposed multi-modal microscopy is used to study inflammation caused by lipopolysaccharide (LPS) on rat macrophages. The TIRF microscopy showed that LPS inflammatory molecule disrupts the cell membrane and causes cells to significantly expand across a substrate. While, QPI module quantified changes in the sub-cellular content of the LPS challenged macrophages, showing a net decrease in its maximum phase values.

Multi-modal chip-based fluorescence and quantitative phase microscopy for studying inflammation in macrophages

In this work, we demonstrate single-shot, simultaneous recording and subsequent retrieval of one incoherent and two holographic (intensity and phase) images from the same camera frame. Demultiplexing of incoherent and holographic signals in the spatial frequency domain is made possible by carrier frequency modulation and spatial oversampling intrinsic to the off-axis digital holographic configuration. In particular, we show applications to combined fluorescence and digital holographic microscopy, as well as combined bright field and holographic second harmonic generation microscopy.

Single‐shot, simultaneous incoherent and holographic microscopy

We apply multiple-wavelength digital holography in combination with epi-fluorescence microscopy in order to generate quantitative phase and fluorescence information from cell samples. While digital holography provides high precision morphological information, the addition of fluorescence supplies the specificity needed to identify cellular constituents. By the application of multiple-wavelength digital holography it is possible to obtain the complete wavefront data deterministically and in real-time. We demonstrate this dual-mode imaging capability through the investigation of living cells.

Dual modality live cell imaging with multiple-wavelength digital holography and epi-fluorescence

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Dual modality live cell imaging with multiple-wavelength digital holography and epi-fluorescence